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We examine feedstock cost and minimum selling price for ethanol production from corn stover as a function of scale, stover yield, participation rate, and price incentives for two conversion technologies: a conventional base case featuring thermochemical pretreatment with added cellulase, and an advanced case featuring consolidated bioprocessing with cotreatment (C‐CBP). Delivered feedstock cost ranged from $85 Mg−1 at small (10 million gallons year−1 or ~38 million L year−1) scale with high yield and participation rates to $124 Mg−1 at large scale (60 million gallons year−1 or 227 million L year−1) and low yield and participation rates. The minimum ethanol selling price (MESP) was approximately twofold lower for the advanced case compared with the base case. The payback period was several times lower for the advanced case compared with the base case, with increasing disparity at smaller scales, and was highly sensitive to ethanol price supports. For both C‐CBP and the conventional processing paradigm, MESP decreased with increasing scale, indicating that the cost penalty due to higher feedstock transport distances was more than outweighed by lower capital costs. However, the cost penalty for operation at small scale, expressed in $ gallon−1 ethanol, is lower for C‐CBP than for the conventional paradigm by roughly twofold. Particularly for initial applications of C‐CBP, we speculate that this cost penalty will likely be modest compared with the anticipated benefits of small‐scale operation such as increased opportunity to use existing infrastructure, easier plant siting and supply chain establishment, and lower total investment required. [ABSTRACT FROM AUTHOR]

Corn cobs consist primarily of a lignocellulosic material comprising hemicellulose, cellulose, and lignin in a crystalline state, which is resistant to microbial saccharification. Bioethanol production from corn cobs has rarely been attempted, especially using chemical pretreatment methods. The present study deals with the production and purification of fungal (Phanerochaete chrysosporium MTCC 787 and Pleurotus florida PAU 22‐01) extracellular ligninolytic enzymes – lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac) – followed by their utilization for the biological pretreatment of corn cobs along with saccharification using commercial cellulase. Crude LiP, MnP, and Lac demonstrated specific activity of 2.23, 2.1, and 2.63 U/mg, respectively. The one‐step purification of crude enzyme using diethyl amino ethyl (DEAE) cellulose ion exchange chromatography resulted in 11.3, 10.1 and 8.62‐fold purification of LiP, MnP and Lac activity, respectively, with corresponding specific activity of 25.1 U/mg (LiP), 21.2 U/mg (MnP) and 22.7 U/mg (Lac) in the partially purified ligninozymes. Using the latter, biological pretreatment of 2.5 g corn cobs in a reaction volume of 30 mL containing approximately 200 units of Lac, Lip and MnP enzymes (in phosphate buffer, pH 6) resulted in a maximum of 78.4% delignification with a saccharification efficiency of 97.1% using commercial cellulases. [ABSTRACT FROM AUTHOR]

The high cost of cellulolytic enzyme complexes (CECs) has been a significant impediment to the commercial production of bioproducts from lignocellulose biomass. This study aimed to develop a cost‐effective CEC derived from Penicillium ucsense (former Penicillium echinulatum), utilizing diverse forms of pretreated sugarcane bagasse as the primary carbon/inductor source. Among the different pretreatments used, the hydrothermal pretreatment followed by NaOH delignification (BHD) produced higher FPase and xylanase activities (4.5 FPU mL–1 and 120 IU mL–1) in bioreactor experiments at 20 g BHD L–1 initial concentration. A batch‐mode assay conducted across a range of initial carbon source (5 to 60 g L–1) confirmed the highest FPase activity (4.0 to 5.0 FPU mL–1 at 120 h), in the range of 20–40 g BHD L–1. During these assays the agitation rate, controlled by dissolved O2, tended to stabilize at lower levels, indicating substrate limitation. Conversely, higher initial carbon source concentrations led to an excess of glucose, likely triggering carbon catabolite repression and inhibiting cellulase production. This insight prompted the development of a controlled pulsed fed‐batch strategy, resulting in FPase activity of 11 FPU mL–1 at 220 h using 90 g L–1 BHD controlled fed into the bioreactor. An enzymatic hydrolysis procedure using the generated CEC was also optimized using a central composite rotational design (CCRD). The optimized enzyme hydrolysis conditions achieved a reducing sugar concentration of 80.9 g L–1 in 48 h using 170 g L–1 of BHD as the substrate at a ratio of 15 FPU of enzyme substrate per g of BHD. A preliminary economic assessment demonstrated that, for a first‐ and second‐generation (1G + 2G) ethanol biorefinery, the cost contribution of enzymes would be about US$0.2/L of biofuel. In conclusion, an efficient and highly productive procedure was developed successfully for the production of a CEC. It was particularly effective for the enzymatic hydrolysis of pretreated sugarcane bagasse. [ABSTRACT FROM AUTHOR]

Lignocellulosic biomass, the most abundant natural resource on earth, can be used for cellulosic ethanol production but requires a pretreatment to improve enzyme access to the polymeric sugars while obtaining value from the other components. γ‐Valerolactone (GVL) is a promising candidate for biomass pretreatment since it is renewable and bio‐based. In the present work, the effect of a pretreatment based on GVL on the enzymatic saccharification of white birch was evaluated at a laboratory scale and the importance of the washing procedure for the subsequent saccharification was demonstrated. Both the saccharification yield and the production of cellulosic ethanol were higher using a noncommercial enzyme crude from Talaromyces amestolkiae than with the commercial cocktail Cellic CTec2 from Novozymes. Furthermore, the production of extracellular cellulases by T. amestolkiae has been optimized in 2 L bioreactors, with improvements ranging from 40% to 75%. Finally, it was corroborated by isoelectric focus that optimization of cellulase secretion by T. amestolkiae did not affect the pattern production of the main β‐glucosidases and endoglucanases secreted by this fungus. [ABSTRACT FROM AUTHOR]

Cellulose reducing ends are believed to play a vital role in the cellulose recalcitrance to enzymatic conversion. However, their role in insoluble cellulose accessibility and hydrolysis is not clear. Thus, in this study, reducing ends of insoluble cellulose derived from various sources were modified by applying reducing and/or oxidizing agents. The effects of cellulose reducing ends modification on cellulose reducing ends, cellulose structure, and cellulose accessibility to cellulase were evaluated along with the impact on cellulose hydrolysis with complete as well purified cellulase components. Sodium borohydride (NaBH4) reduction and sodium chlorite‐acetic acid (SC/AA) oxidation were able to modify more than 90% and 60% of the reducing ends, respectively, while the bicinchoninic acid (BCA) reagent applied for various cycles oxidized cellulose reducing ends to various extents. X‐ray diffractograms of the treated solids showed that these treatments did not change the cellulose crystalline structure and the change in crystallinity index was insignificant. Surprisingly, it was found that the cellulose reducing ends modification, either through selective NaBH4 reduction or BCA oxidation, had a negligible impact on cellulose accessibility as well on cellulose hydrolysis rates or final conversions with complete cellulase at loadings as low as 0.5 mg protein/g cellulose. In fact, in contrast to what is traditionally believed, modifications of cellulose reducing ends by these two methods had no apparent impact on cellulose conversion with purified cellulase components and their synergy. However, SC/AA oxidation resulted in significant drop in cellulose conversion (10%–50%) with complete as well purified cellulase components. Nonetheless, further research revealed that the cause for drop in cellulose conversion for the SC/AA oxidation case was due to primary hydroxyl groups (PHGs) oxidation and not the oxidation of reducing ends. Furthermore, it was found that the PHGs modification affects cellulose accessibility and slows the cellulase uptake as well resulting in significant drop in cellulose conversions. [ABSTRACT FROM AUTHOR]

Cyanobacteria and heterotrophic bacteria show many similarities in their structure but are different based on their nutritional habits. Autotrophic cyanobacteria and heterotrophic bacteria coexist closely in their natural environments and are capable of a variety of interactions (synergistic, neutral, and antagonistic) due to the secretion of growth‐promoting/retarding (toxin) substances by the bacterial partner. In this study, the cyanobacteria Lyngbya aestuarii and its associated bacteria Alcaligenes aquatilis isolated from the mangrove water of Bhitarkanika showed an antagonistic interaction. The growth of axenic and co‐cultured (1:1; cyanobacteria to bacteria) cyanobacteria was examined under photoautotrophic conditions, to evaluate the interactions between the two groups. The bacteria were found to have cellulase and protease activity and were able to obstruct the growth of cyanobacteria under co‐culture conditions. Besides that, the bacteria secrete many allelochemicals like phenol, tannin, amino acids (alanine and aspartic acid), and hydroxamate siderophore to its surroundings which inhibit the growth and metabolism of the cyanobacteria under co‐cultured conditions. Findings from this study indicate its possible application for wastewater management. [ABSTRACT FROM AUTHOR]

Because of the association with other complex polysaccharides, extracting and utilizing cellulose from lignocellulosic materials requires the combined action of a broad range of carbohydrate‐active enzymes, including multiple glycoside hydrolases (GHs) and lytic polysaccharide monooxygenases (LPMOs). The interplay between these enzymes and the way in which Nature orchestrates their co‐existence and combined action are topics of great scientific and industrial interest. To gain more insight into these issues, we have studied the lignocellulose‐degrading abilities of an enzyme from Caldibacillus cellulovorans (CcLPMO10‐Man5), comprising an LPMO domain, a GH5 mannanase domain and two family 3 carbohydrate‐binding modules (CBM3). Using a natural softwood substrate, we show that this enzyme promotes cellulase activity, i.e., saccharification of cellulose, both by removing mannan covering the cellulose and by oxidatively breaking up the cellulose structure. Synergy with CcLPMO10‐Man5 was most pronounced for two tested cellobiohydrolases, whereas effects were smaller for a tested endoglucanase, which is in line with the notion that cellobiohydrolases and LPMOs attack the same crystalline regions of the cellulose, whereas endoglucanases attack semi‐crystalline and amorphous regions. Importantly, the LPMO domain of CcLPMO10‐Man5 is incapable of accessing the softwood cellulose in absence of the mannanase domain. Considering that LPMOs not bound to a substrate are sensitive to autocatalytic inactivation, this intramolecular synergy provides a perfect rationale for the evolution of modular enzymes such as CcLPMO10‐Man5. The intramolecular coupling of the LPMO with a mannanase and two CBMs ensures that the LPMO is directed to areas where mannans are removed and cellulose thus becomes available. [ABSTRACT FROM AUTHOR]

Heavy metals contamination poses a significant environmental threat, which requires the development of eco‐friendly bioremediation techniques. Present research work was conducted using wheat straw in submerged conditions for the growth of white rot fungus, Phlebia brevispora in the presence of varying concentrations of Cd, Pb, Ni, and Cr. The lignocellulolytic enzyme production ability of fungus was monitored. A wheat straw‐based fixed‐bed bioreactor was designed to treat metal‐containing water in continuous mode. The fungus showed a positive influence on laccase production, cellulase production, and lignin peroxidase activity. As observed through atomic absorption spectroscopy (AAS), Cr removal efficiency was 78%–80%, regardless of the initial metal concentration. The Cr was also found on the surface of fungal mycelium as per the results obtained from SEM–EDX. The continuous bioreactor achieved 98%–99% metal removal, making it a natural and cost‐effective solution for metal removal from wastewater using P. brevispora. [ABSTRACT FROM AUTHOR]

Cellulases play an important role in the bioconversion of lignocellulose. Microorganisms found in extreme environments are a potentially rich source of cellulases with unique properties. Due to the uniqueness of the environment, the abundant microbial resources in the Qinghai–Tibet Plateau (QTP) are worth being explored. The aim of this study was to isolate and characterize an acidic, mesophilic cellulase‐producing microorganism from QTP. Moreover, the fermentation conditions for cellulase production were optimized for future application of cellulase in the development of lignocellulose biomass. A novel cellulase‐producing strain, Penicillium oxalicum XC10, was isolated from the soil of QTP. The cellulase produced by XC10 was a mesophilic cellulase that exhibited good acid resistance and some cold‐adaptation properties, with maximum activity at pH 4.0 and 40°C. Cellulase activity was significantly enhanced by Na+ (p < 0.05) and inhibited by Mg2+, Ca2+, Cu2+, and Fe3+ (p < 0.05). After optimization, maximum cellulase activity (8.56 U/mL) was increased nearly 10‐fold. Optimal fermentation conditions included an inoculum size of 3% (v/v) in a mixture of corn straw (40 g/L), peptone (5 g/L), and Mg2+ (4 g/L), at pH 4.0, 33°C, and shaking at 200 rpm. The specific properties of the P. oxalicum XC10 cellulase suggest the enzyme may serve as an excellent candidate for the bioconversion and utilization of lignocellulose biomass generated as agricultural and food‐processing wastes. [ABSTRACT FROM AUTHOR]

Introduction: Efficient extraction of camptothecin (CPT), an anticancer agent from the commercial source Nothapodytes nimmoniana (J. Graham) Mabb in India, is of paramount importance. CPT is present in the highest concentration in the stem portion, and the stem can be readily harvested without uprooting the plant. The fluorescence microscopy mapping of the bark matrix for CPT revealed its presence in a free form within both the outer (epidermal and cortical tissues) and inner (xylem and phloem tissues) sections. The bark matrix primarily consists of cellulose, hemicellulose, and lignin, rendering it woody, rigid, and resistant to efficient solvent penetration for CPT extraction. We proposed a hypothesis that subjecting it to disruption through treatment with hydrolytic enzymes like cellulase and xylanase could enhance solvent diffusion, thereby enabling a swift and effective extraction of CPT. Objective: The present study was aimed at enzyme‐assisted extraction, using cellulase and xylanase for hydrolytic disruption of the cells to readily access CPT from the stem of the plant N. nimmoniana (J. Graham) Mabb. Methodology: The hydrolytic cell disruption of ground powder from the stem bark was studied using cellulase and xylanase enzymes. The enzymatically pretreated stem bark powder was subsequently recovered by filtration, dried, and subjected to extraction with methanol to isolate CPT. This process was optimised through a Box–Behnken design, employing a one‐factor‐at‐a‐time approach to assess parameters such as enzyme concentration (2–10% w/w), pH (3–7), incubation time (6–24 h), and solid‐to‐solvent ratio (1:30–1:70 g/mL). CPT was characterised using proton nuclear magnetic resonance (1H‐NMR) and Fourier transform infrared (FTIR) spectra, and a high‐performance liquid chromatography (HPLC) method was developed for quantification. Results: The cellulase and xylanase treatment resulted in the highest yields of 0.285% w/w and 0.343% w/w, with efficiencies of 67% and 81%, respectively, achieved in a significantly shorter time compared to the untreated material, which yielded 0.18% with an efficiency of only 42%. Extraction by utilising the predicted optimised process parameters, a nearly two‐fold increase in the yield, was observed for xylanase, with incubation and solvent extraction times set at 16 and 2 h, respectively. Scanning electron microscopy (SEM) images of the spent material indicated perforations attributed to enzymatic action, suggesting that this could be a primary factor contributing to the enhanced extraction. Conclusion: Enzyme‐mediated hydrolytic cell disruption could be a potential approach for efficient and rapid isolation of CPT from the bark of N. nimmoniana. Enzyme‐assisted efficient extraction of camptothecin (CPT), an anticancer agent from Nathapoydytes nimmonia, was achieved by disrupting the woody rigid bark matrix through treatment with enzymes cellulase and xylanase. A reference sample of CPT, generated by the extraction of the bark and characterized, was used for quantification employing the HPLC method. Parameters such as enzyme concentration, temperature, pH, solid‐to‐liquid ratio, were investigated and process optimised employing the Box‐Benkhen design resulting in an almost two‐fold increase in yield. [ABSTRACT FROM AUTHOR]

Summary: The increased demand for vegetable oils calls for new strategies for enhancing the yield of mechanical oil extraction, preferably without the use of potentially hazardous solvents such as n‐hexane. Enzyme‐assisted oil extraction has been studied under aqueous conditions, where the oil recovery was enhanced due to the partial enzymatic breakdown of cell wall compounds. However, this process is often not feasible for industrial applications, where mechanical pressing of dry seeds is implemented. This study provides a proof‐of‐concept for enzyme‐assisted rapeseed oil extraction with minimum water addition (1 mL per 100 g rapeseeds). The application of cellulytic and pectolytic enzymes increased the reducing sugar content as a result of the carbohydrate breakdown. The oil recovery in the mechanical pressing was increased significantly from ~60% to ~65%, and the quality of the oils was not significantly affected by the enzyme treatment. [ABSTRACT FROM AUTHOR]

Pestalotiopsis guepinii is a pathogenic fungus that causes grey blight on Camellia pitardii. In this study, we investigated the enzyme activity and kinetics of these cell‐wall‐degrading enzymes (CWDEs) produced by Pestalotiopsis guepinii in both C. pitardii leaves and culture medium. Our enzyme activity experiments revealed that the activities of xylanase, pectin methyl‐galacturonase (PMG), β‐1,4‐endoglucanase (Cx), and β‐glucosidase were high in both C. pitardii leaves and culture medium. These enzymes played a significant role in the pathogenic process. However, the activity of laccase was found to be very low and had a minor impact on the pathogenic process. Furthermore, our enzyme dynamics experiments demonstrated that the optimal reaction temperature for PMG and Cx was 50°C, while for β‐glucosidase and xylanase, it was 60°C. The optimal reaction pH for Cx, β‐glucosidase, and xylanase was 5.0, whereas for PMG, it ranged from 5.0 to 6.0. This indicates that these four enzymes prefer acidic conditions. Moreover, we observed that the activities of Cx, PMG, and xylanase decreased with increasing reaction time. On the other hand, the activity of β‐glucosidase initially increased sharply and then decreased slowly. The maximum reaction rates of the four cell‐wall‐degrading enzymes were ranked as follows: xylanase > PMG > β‐glucosidase > Cx. Additionally, the affinities of these enzymes with substrates were ranked as follows: PMG < Cx < xylanase < β‐glucosidase. [ABSTRACT FROM AUTHOR]

Two methods for extraction of oil from inajá pulp were studied; an enzymatic extraction, which was evaluated using the central composite rotational design (CCRD) combined with the response surface methodology (RSM), and the conventional solvent extraction. The enzymatic extraction method had no significant effect (p > 0.05%) on the fatty acid profile of inajá oil, which was 45.7% monounsaturated, 10.8% polyunsaturated, and 43.6% saturated. Oleic, palmitic, linoleic, and myristic acids were the predominant fatty acids. The enzymatic extraction method interfered with the total phenolic compounds, which were lower (p < 0.05%) than the total phenolic compounds obtained in the inajá pulp oil extracted by solvent. On the other hand, the carotenoid content was higher in the inajá pulp oil obtained by the enzymatic extraction (p < 0.05%). The enzymatic extraction showed an efficiency equivalent to that of the solvent extraction, and resulted in an oil that contains superior nutritional properties. Green technologies, such as enzymatic extraction for the production of vegetable oils, can be used to replace conventional methods that use solvent, thereby reducing environmental impact and the operating cost of the process itself. In the enzymatic extraction, the oil is obtained without the need for a solvent separation step. Moreover, this method of extraction produces an aqueous fraction and a solid residue (defatted pulp), both of which can be used as by‐products with potential for application in the food industry and, therefore, generate greater added value. [ABSTRACT FROM AUTHOR]

BACKGROUND: The high fibre content of whole plants of Broussonetia papyrifera limits its efficient utilization. Ferulic acid esterase (FAE), in combination with xylanase, can effectively cleave the lignin–carbohydrate complex, promoting the function of cellulase. However, little is known about the impact of these additives on silage. To effectively utilize natural woody plant resources, FAE‐producing Lactiplantibacillus plantarum RO395, xylanase (XY) and cellulase (CE) were used to investigate the dynamic fermentation characteristics, fibre and nitrogen components and microbial community structure during B. papyrifera ensiling. RESULTS: Broussonetia papyrifera was either not treated (CK) or treated with FAE‐producing lactic acid bacteria (LP), CE, XY, LP + CE, LP + XY or LP + CE + XY for 3, 7, 15, 30 or 60 days, respectively. In comparison with those in the CK treatment, the L. plantarum and enzyme treatments (LP + CE, LP + XY and LP + XY + CE), especially the LP + XY + CE treatment, significantly increased the lactic acid concentration and decreased the pH and the contents of acid detergent insoluble protein and NH3‐N (P < 0.05). Enzyme addition improved the degradation efficiency of lignocellulose, and a synergistic effect was observed after enzyme treatment in combination with LP; in addition, the lowest acid detergent fibre, neutral detergent fibre, hemicellulose and cellulose contents were detected after the LP + CE + XY treatment (P < 0.05). Moreover, CE, XY and LP additions significantly improved the microbial community structure, increased the relative abundance of Lactiplantibacillus and Firmicutes, and effectively inhibited undesirable bacterial (Enterobacter) growth during ensiling. CONCLUSION: FAE‐producing L. plantarum and the two tested enzymes exhibited synergistic effects on improving the quality of silage, which indicates that this combination can serve as an efficient method for improved B. papyrifera silage utilization. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND: Cherry tomatoes are nutritious and favored by consumers. Processing them into dried cherry tomatoes can prolong their storage life and improve their flavor. The pretreatment of tomato pericarp is crucial for the subsequent processing. However, the traditional physical and chemical treatments of tomato pericarp generally cause nutrient loss and environmental pollution. RESULTS: In this study, a novel enzymatic method for cherry tomatoes was performed using mixed enzymes containing cutinase, cellulase and pectinase. Results showed that the pericarp permeability of cherry tomatoes was effectively improved due to enzymatic treatment. Changes in the microscopic structure and composition of the cuticle were revealed. After treatment with different concentrations of enzymes, cherry tomatoes exhibited higher pericarp permeability and sensory quality to varying degrees. The lycopene content and total polyphenol content significantly increased 2.4- and 1.45-fold, respectively. In addition, the satisfactory effect of the six-time reuse of enzymes on cherry tomatoes could still reach the same level as the initial effect, which effectively reduced the cost of production. CONCLUSIONS: This study revealed for the first time that a mixed enzymatic treatment consisting of cutinase, pectinase and cellulase could effectively degrade the cuticle, enhance the pericarp permeability and improve the quality of cherry tomatoes, with the advantages of being mildly controllable and environmentally friendly, providing a new strategy for the processing of dried cherry tomatoes. [ABSTRACT FROM AUTHOR]

Summary: The integration of carbon dioxide‐nanobubbles (CO2‐NBs) with cellulase has led to the development of an innovative extraction system for phenolic compound retrieval from P. cyrtonema (Polygonatum cyrtonema) Hua rhizomes. Four critical factors influencing on CO2‐NBs‐cellulase were methodically investigated and optimised through single‐factor experiments. The optimal experimental conditions are 30 cycles, 0.35 MPa, pH 5.45 and 0.025 g/mL cellulase aqueous solution. The stability of system was characterised meticulously, indicating the viability for at least 24 h. Comparative assessment of bioactive constituents and antioxidant activity demonstrated exceptional extraction efficiency and potential synergistic effects of CO2‐NBs‐cellulase. Synergistic effects and reaction mechanisms during extraction were explored using Fourier‐transform infrared spectroscopy, differential scanning calorimetry and scanning electron microscopy, which contributed to mechanism understanding with significant alterations in partial structure and heightened thermal stability we found. In essence, this study introduces an eco‐friendly, energy‐efficient extraction system paradigm based on a novel nanobubble‐enzyme binding approach. [ABSTRACT FROM AUTHOR]

Strawberry (Fragaria × ananassa) is one of the most important fruit crops, worldwide. In June 2023, strawberry fruits with pink mould symptoms were collected from a storage facility in Khorasan Razavi province of Iran. Trichothecium roseum was isolated from symptomatic fruit and identified through morphological characteristics. Molecular identification was carried out based on the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) using ITS1 and ITS4 primers. Pathogenicity was confirmed according to Koch's postulates. Cell wall–degrading enzyme secretion was investigated as a potential factor in fungal pathogenicity. The results of morphological identification, ITS analysis and pathogenicity test indicated that the causal agent was T. roseum. Enzyme activity assays revealed that T. roseum obtained from strawberry with a high level of aggressiveness was able to secrete cellulase and pectinase. To our knowledge, this is the first report of post‐harvest pink mould of strawberry caused by T. roseum in Iran. [ABSTRACT FROM AUTHOR]

Sugarcane bagasse (SB) is an abundantly available agrowaste that is used extensively as a substrate for the production of different industrial enzymes. It is a renewable resource, rich in fermentable sugars. However, the presence of lignin makes it difficult for microbes to degrade it and so it is difficult to utilize it. Keeping in mind the role of white rot fungi in lignin degradation, Trametes pubescens MB89 was used in this study for the delignification of SB with the simultaneous production of laccase. The fungus was also used for the degradation of malachite green dye. A Box–Behnken design (BBD) was used to optimize variables affecting solid state fermentation of SB for laccase production. The leftover SB after fermentation was used for cellulase production by the thermophilic bacterium Neobacillus sedimentimangrovi UE25. The BBD depicted optimized conditions as 60% moisture, particle size 100 μ, 10 days' incubation period, 450 μM CuSO4, and a temperature of 30 °C. The laccase titers of 1271 IU mL−1 were obtained under optimized conditions. Neobacillus sedimentimangrovi UE25 produced 4.574 IU mL−1 of endoglucanase and 1.812 IU mL−1 of β‐glucosidase by utilizing fungal pretreated SB. Scanning electron microscopy and Fourier transform infrared spectroscopy confirmed the impact of fungal cultivation on SB. Pores were observed after microbial pretreatment and there were changes in the morphological characteristics such as the pith and fiber of SB. To test the ability of laccase to break down aromatic dyes, we grew the fungus T. pubescens MB89 in a nutrient‐rich medium supplemented with malachite green dye. The fungus was able to remove 73.68% of the malachite green dye from the medium in 24 h. This shows that T. pubescens MB89 has a range of potential applications for environmental and industrial purposes. [ABSTRACT FROM AUTHOR]

Here we describe the first crystal structure of a beta‐1,4‐endoglucanase from a brown‐rot fungus, Gloeophyllum trabeum GtCel45A, which belongs to subfamily C of glycoside hydrolase family 45 (GH45). GtCel45A is ~ 18 kDa in size and the crystal structure contains 179 amino acids. The structure is refined at 1.30 Å resolution and Rfree 0.18. The enzyme consists of a single catalytic module folded into a six‐stranded double‐psi beta‐barrel domain surrounded by long loops. GtCel45A is very similar in sequence (82% identity) and structure to PcCel45A from the white‐rot fungus Phanerochaete chrysosporium. Surprisingly though, initial hydrolysis of barley beta‐glucan was almost twice as fast in GtCel45A as compared to PcCel45A. [ABSTRACT FROM AUTHOR]

BACKGROUND: This work aimed to study different methodologies for extracting enzymes produced by the endophytic fungi isolated from mangrove Neofusicoccum parvum, Penicillium glabra series and Bjerkandera sp. grown on organic rice. The enzymatic activities of amylase, cellulase, laccase, lipase, peroxidase, protease and keratinase were evaluated for future applications in biotechnology and enzymology. The evaluation of the extraction was carried out using the design of experiments methodology, where variations were made in pH (4–8), temperature (20–36 °C) and agitation (150–200 rpm), in times of 1, 6 and 12 h. RESULTS: Among the enzymes analyzed, promising activities were found for amylase, cellulase, lipase, protease, peroxidase and especially keratinase, presenting activities of up to 7049.47 U g−1 in the fermentation carried out with the fungus Neofusicoccum parvum, 3538.52 U g−1 with the fungus Penicillium glabra series and 5068.20 U g−1 with the fungus Bjerkandera sp. This indicates that the fungi are promising for the production of multienzyme cocktails. CONCLUSION: It should be emphasized that the microorganisms and extraction conditions evaluated resulted in a series of promising results, that is, in the case of aiming to obtain different enzymes by other extraction conditions, after fermentation of various microorganisms on rice, several options of situations are presented and validated in this work, which allows the choice of the same, depending on the future application potential of these biocatalysts. © 2023 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

The yeast expression system, Pichia pastoris, is one of the most robust and versatile expression systems in biotechnology and molecular biology, generally considered as a safe host for heterologous protein expression, especially for producing cellulases and xylanases at the industrial level. Despite the high recombinant protein expression rate, the potential of the P. pastoris expression system has still not been fully explored. Cultivation of P. pastoris under optimized conditions greatly relies on the strain and is associated with certain problems such as promoter strength, sensitivity to methanol, and oxygen demand. To address these issues, different genetic engineering strategies have been employed. Advancements in promoter engineering, optimization of gene dosage and codon usage, recombinant plasmid engineering using CRISPR/Cas9 system, and directed evolution strategies have proven beneficial to the yield of cellulase expression levels. This study will systemically review recent progress in various genetic engineering strategies to enhance cellulase and xylanase expression in the P. pastoris expression system. The utilization of alcohol oxidase 1 promoter (pAOX1), methanol‐free system, and recombinant plasmid engineering for improved production of these enzymes are highlighted. Additionally, we discuss the recent advancements in the P. pastoris expression system toolbox for improved cellulase and xylanase production, thus providing a deep insight into how P. pastoris is becoming the indispensable platform for heterologous protein production. © 2023 Society of Chemical Industry (SCI). [ABSTRACT FROM AUTHOR]

Background: Pesticide formulations based on nanotechnology can effectively improve the efficiency of pesticide utilization and reduce pesticide residues in the environment. In this study, mesoporous silica nanoparticles containing disulfide bonds were synthesized by the sol–gel method, carboxylated and adsorbed with lufenuron, and grafted with cellulose to obtain a lufenuron‐loaded nano‐controlled release formulation (Luf@MSNs‐ss‐cellulose). Results: The structure and properties of Luf@MSNs‐ss‐cellulose were characterized. The results showed that Luf@MSNs‐ss‐cellulose exhibits a regular spherical shape with 12.41% pesticide loading. The highest cumulative release rate (73.46%) of this pesticide‐loaded nanoparticle was observed at 7 days in the environment of glutathione and cellulase, which shows redox‐enzyme dual‐responsive performance. As a result of cellulose grafting, Luf@MSNs‐ss‐cellulose had a small contact angle and high adhesion work on corn leaves, indicating good wetting and adhesion properties. After 14 days of spraying with 20 mg L−1 formulations in the long‐term control efficacy experiment, the mortality of Luf@MSNs‐ss‐cellulose against Ostrinia furnacalis larvae (56.67%) was significantly higher than that of commercial Luf@EW (36.67%). Luf@MSNs‐ss‐cellulose is safer for earthworms and L02 cells. Conclusion: The nano‐controlled release formulation obtained in this study achieved intelligent pesticide delivery in time and space under the environmental stimulation of glutathione and cellulase, providing an effective method for the development of novel pesticide delivery systems. © 2023 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Global production of sugarcane bagasse (SB) by sugar industries exceeds more than 100 tons per annum. SB is rich in lignin and polysaccharide and hence can serve as a low‐cost energy and carbon source for the growth of industrially important microorganism. However, various other applications of SB have also been investigated. In this study, SB was used as an adsorbent to remove an azo dye, malachite green. Subsequently, the dye‐adsorbed SB was fermented by Trametes pubescens MB 89 for the production of laccase enzyme. The fungal pretreated SB was further utilized as a substrate for the simultaneous production of multiple plant cell wall degrading enzymes including, cellulase, xylanase, pectinase, and amylase by thermophilic bacterial strains. Results showed that 0.1% SB removed 97.04% malachite green at 30°C after 30 min from a solution containing 66 ppm of the dye. Fermentation of the dye‐adsorbed SB by T. pubescens MB 89 yielded 667.203 IU mL−1 laccase. Moreover, Brevibacillus borstelensis UE10 produced 38.41 and 18.6 IU mL−1 β‐glucosidase and pectinase, respectively, by using fungal‐pretreated SB. Cultivation of B. borstelensis UE27 in the medium containing the same substrate yielded 32.14 IU mL−1 of endoglucanase and 27.23 IU mL−1 of β‐glucosidase. Likewise, Neobacillus sedimentimangrovi UE25 could produce a mix of β‐glucosidase (37.24 IU mL−1), xylanase (18.65 IU mL−1) and endoglucanase (26.65 IU mL−1). Hence, this study led to the development of a method through which dye‐containing textile effluent can be treated by SB along with the production of industrially important enzymes. [ABSTRACT FROM AUTHOR]

Advances in biodigester technology have been promoted by the growing study of anaerobic digestion, with the aim of improving processes related to the utilized waste load. This research used the statistical tool Central Composite Rotatable Design to optimize the combination of microorganisms and enzymes for biodigestion. The experiment involved testing two microorganisms (EA and EB) and five enzymes (amylase, protease, lipase, cellulase, and amyloglucosidase) with food waste from nearby restaurants and WWTP sludge as an inoculant. The experiment was set up as Central Composite Rotatable Design (CCRD) 26‐1, with 50 trials and a concentration of 14.5 g of volatile solids per liter. The dependent variables of cumulative biogas production (BG) and cumulative biomethane production (BM) were evaluated. The results showed that the independent variables EB and CA had negative and significant effects on BG and BM production. With optimized process conditions, the bioaccelerator significantly improved BG and BM responses compared to control trials by 72% and 88%, respectively. The biodigestion technique is a viable option for the treatment of organic waste, and the use of bioaccelerators may be a promising approach to improving biodigestion processes. This study contributes to the ongoing research on anaerobic digestion and biodigester technology, which seeks to improve the sustainable management of organic waste while producing renewable energy. [ABSTRACT FROM AUTHOR]

Summary: The addition of wheat bran dietary fibre to the pasta improves its nutritional value, while affects negatively the quality of the product. This study aimed to improve the physicochemical and sensorial properties of wheat bran pasta by evaluating the effect of cellulase‐treated wheat bran (CTWB) on the rheological properties of the dough and the quality of produced pasta. Untreated wheat bran was used to produce control samples. Then, wheat bran at levels of 5, 10, 15, and 20% was treated with 0.1 and 0.3% cellulase. Rheological characteristics of the produced dough were measured by farinograph and extensograph. Finally, the cooking and sensory properties of pasta were evaluated. Based on the obtained results, applying CTWB has led to an increase in farinograph quality number, an increase in dough energy and extensibility, and a decrease in the resistance to extension of the dough. Also, the cooking quality (cooking loss and adhesiveness) of pasta samples improved using CTWB. Pasta samples containing 5 and 10% wheat bran treated with 0.3% cellulase were selected as optimum treatments for both dough and pasta properties. The findings in this study indicated that CTWB added to pasta improved the cooking characteristics and sensorial properties of pasta, significantly. [ABSTRACT FROM AUTHOR]

Summary: Hemp milk is a plant‐based beverage emerging as a sustainable dairy alternative and it is essential to improve the emulsification properties and stability of hemp protein for increasing acceptability. This study demonstrates the preparation of protein‐rich hemp milk with enhanced emulsion stability and functional properties through cellulase hydrolysis of fibre components in hemp protein. The emulsions were prepared using the hemp dispersions obtained after cellulase treatment followed by mechanical treatment (ME) and in combination with either enzymatic (trypsin) (EE) or mild chemical (pH‐shift) treatment (PE). The influences of these treatments on the particle size distribution, zeta potential, flow behaviour, microstructure, flocculation, and coalescence properties of the emulsions are investigated. The emulsions (EE/PE) prepared with dispersions processed through tryptic or pH shift treatments showed improved stability as indicated by reduced flocculation and coalescence indices (8.9% and 10.8%, for EE and 11.8% and 13.2% for PE) as compared to only mechanically treated emulsion (ME) (37.5% and 15.3%). Furthermore, the enhanced stability of EE and PE samples is attributed to a reduction in particle size and enhanced zeta potential (EE: 45.1 mV > PE: 38.7 mV > ME: 34.1 mV). [ABSTRACT FROM AUTHOR]

Cellobiose dehydrogenase (CDH) catalyzes the oxidation of cello‐oligosaccharides and transfers electrons to oxygen or other electron receptors, such as lytic polysaccharide oxygenases (LPMOs). Cellobiose dehydrogenase has the potential to be used as an electron donor for LPMOs. Therefore, it is important to obtain CDHs under reaction conditions that are compatible with those of cellulases and LPMOs. Here, a CDH gene from Myceliophthora thermophila (MtCDH‐A) was constitutively expressed in Trichoderma reesei, and its ability to donate electrons to the LPMO was validated. MtCDH‐A has an optimal pH of 6.0 and maintains relatively high activity over a wide pH range. The optimal temperature was 60°C, and it remained stable at 50°C for a long time. These enzymatic properties were compatible with the catalytic conditions of fungal LPMOs and cellulases. When MtCDH‐A was used as an electron donor, the synergistic effect of LPMO and cellulase improved the degradation efficiency of phosphoric acid‐swollen cellulose by 141%. This work provides a novel insight into finding a stable electron donor for LPMOs and a new direction for finding more efficient thermostable CDHs in newly identified fungal species. © 2023 Society of Industrial Chemistry and John Wiley & Sons Ltd. [ABSTRACT FROM AUTHOR]

Aspergillus species have been highlighted in enzyme production looking for industrial applications, notably, amylases are one of the most interesting enzymes. They are capable of hydrolyzing α‐glycosidic linkages of starch and widely used in industrial processes to produce ethanol, glucose, and fructose syrup as well as in the textiles, detergents, and paper industries applications. In this context, this work aimed at the biochemical characterization of the glucoamylase from Aspergillus japonicus and its application in the bio‐bleaching process of recycled paper. The optimum temperature and pH for the glucoamylase assay were standardized as 50°C and 5.5. After 1 h of incubation, glucoamylase retained 90% of its activity at 30–50°C. It also kept 70% of its activity in the pH range of 4.0–6.5 after an hour of incubation. The enzyme led to an increase of 30% in the relative whiteness of 10 dry grams of sulfite paper and magazine paper when applied along with commercial cellulase and 10 mM MnCl2. In addition, after the treatments, the glucoamylase recovered activity was 30%–32%, which indicates a prolonged availability of the enzyme and can considerably curtail the redundant downstream process of the recycled paper bio‐bleaching. Thus, the glucoamylase from A. japonicus has a significant role in bio‐bleaching recycled paper, reducing the necessity of hard chemicals, and improving the industrial process in an interesting economic and ecological mode. [ABSTRACT FROM AUTHOR]

Summary: Citrus processing waste (CPW) accumulates in vast amounts worldwide during fruit processing and its disposal through landfilling is environmentally damaging. However, CPW is also a renewable source of economically and medically relevant compounds such as pectin. There is a need for new methods of extracting pectin from CPW, as conventional methods are unsustainable. Therefore, this study proposed a new environmentally friendly method, which combines ultrasound cavitation with cellulase preparation, Celluclast) into one extraction procedure (UAE), and compared it to conventional acid extraction. The yield and productivity of each method were measured and the extracts were analysed for composition and structural characteristics. UAE and acid extraction pectin yields (26.9 vs. 22.1%) and productivity (0.10 vs. 0.09%/min) values were comparable. The extracts consisted mainly of galacturonic acid and galactose residues, but the ratio was different for each method; 1: 0.34 and 0.84: 1 for acid and UAE‐extracted pectin, respectively. FTIR was used to estimate the degree of esterification of the pectin extracts; 8% and 55% for acid and UAE‐extracted pectin, respectively. TGA revealed a higher ash content in UAE pectin, while XRD revealed similar degrees of polymer crystallinity. Viscosimetry revealed that the acid pectin had a higher molecular weight (70.22 kDa) compared to UAE pectin (197.78 kDa). In conclusion, the UAE procedure is a viable alternative to acid extraction. [ABSTRACT FROM AUTHOR]

The yield and properties of banana juice extracted by enzymatic pretreatments using different enzymes and were compared among themselves and with hot water extraction. The treatments that gave higher yield and better properties of the juices were used for winemaking and the properties of the wines were also compared. A comparison of gas chromatography‐mass spectrometry analysis was also done between the juices and the wines. The various treatments were hot water bath extraction and different enzymes (amylase, cellulase, pectinase, and viscozyme) at two concentrations (0.05% and 0.1%). Juice yield, total soluble sugars, pH, titrable acidity, reducing sugars, and transmittance were analyzed. Best results for juice recovery (46.44 ± 0.50%), total soluble solids (14.8 ± 0.01 °Brix), and transmittance (86.93 ± 2.68%) were achieved using pectinase (0.1%), while viscozyme (0.1%) yielded best results for viscosity (1.49 ± 0.09 mPa.s), pH (5.01 ± 0.05), and acidity (0.277 ± 0.01%). The banana juices from these treatments were then used for wine preparation, and the physicochemical parameters were monitored during a 30‐day storage period. Within 20 days during fermentation, a reduction in total soluble solids from 22 °Brix to 6.1 °Brix was observed. Juice from banana pulp can be successfully extracted using the enzymes pectinase and viscozyme, and wine can be prepared from the enzymatically extracted juices. [ABSTRACT FROM AUTHOR]

Jackfruit (Artocarpus heterophyllus Lam) waste (JFW), a lignocellulosic biomass, is a potential raw material that can be tapped to produce energy and valuable products for human use. In the present work, biobutanol production was studied using jackfruit waste. Initially, the jackfruit waste was pretreated using acid and alkaline agents and hydrolyzed to produce fermentable sugars. The hydrolyzed jackfruit waste was fermented using Clostridium acetobutylicum NCIM 2877 to produce biobutanol. Among the studied pretreatments, sulfuric acid pretreatment produced a maximum sugar yield of 84.21 ± 2.6 mggds−1 in the acid liquor. Enzyme hydrolysis of pretreated jackfruit waste produced a total glucose yield of 215.12 ± 13.44 mggds−1 after optimization. After detoxification using activated charcoal, the JFW hydrolysate produced a maximum biobutanol titer and yield of 1.60 ± 0.5 g L−1, and 106.59 ± 5.1 mg/gglucose, respectively. Central composite design optimization of acetone‐butanol‐ethanol fermentation produced a maximum biobutanol titer of 5.6 ± 0.2 g L−1 (250.00 ± 11.4 mg/gglucose) and total solvent titer of 7.28 ± 0.3 g L−1 (325.00 ± 13.1 mg/gglucose). A 31 ton annual biobutanol production from JFW, simulated using SuperPro Designer, revealed the unit production cost and payback time to be $10.49/kg and 8.94 years. [ABSTRACT FROM AUTHOR]

Recent studies have identified a significant number of endogenous cellulase genes in various arthropods, including isopods, allowing them to process hydrocarbons efficiently as a food source. While this research has provided insight into underlying gene‐level processes in cellulose decomposition by arthropods, little is known about the existence and expression of cellulase genes in species from cave environments where carbohydrates are sparse. To investigate whether endogenous cellulase genes are maintained in subterranean species, we sequenced the transcriptomes of two subterranean paraplatyarthrid isopod species from calcrete (carbonate) aquifers of central Western Australia and a related surface isopod species. Seven protein‐coding open‐reading frames associated with endoglucanase genes were identified in all species. Orthology inference analyses, using a wide range of cellulase sequences from available databases, supported the endogenous origin of the putative endoglucanase genes. Selection analyses revealed that these genes are primarily subject to purifying selection in most of the sites for both surface and subterranean isopod species, indicating that they are likely to encode functional peptides. Furthermore, evolutionary branch models supported the hypothesis of an adaptive shift in selective pressure acting on the subterranean lineages compared with the ancestral lineage and surface species. Branch‐site models also revealed a few amino acid sites on the subterranean branches to be under positive selection, suggesting the acquisition of novel adaptations to the subterranean environments. These findings also imply that hydrocarbons exist in subsurface aquifers, albeit at reduced levels, and have been utilized by subterranean isopods as a source of energy for millions of years. [ABSTRACT FROM AUTHOR]

Summary: Garden cress (Lepidium sativum) is valued in Asia and some part of Africa for its seed oil and other parts of plant that retain curative activities and act as a tonic in traditional medicine. The residue left over after the oil and protein extraction is a rich source of bioactive compounds and dietary fibres. The garden cress seed residual fibre (GCRF) contained 99.82% total dietary fibre (TDF), 99.4% insoluble dietary fibre (IDF) and 0.38% soluble dietary fibre (SDF). To improve the industrial applications of the GCRF, an approach was taken to break down the IDF into the SDF following a treatment of enzymatic hydrolysis [cellulase at 2% or 5% (w/w)] and high‐pressure processing (600 MPa/15 min) or a combination of both. The combined treatment had significantly improved the SDF concentration by loosening the surface structure of the GCRF and enhanced the hydration properties. The structural changes were elucidated using the Fourier transform infrared spectroscopy, rheometry and scanning electron microscopy. The Fourier transform infrared spectrum revealed a major shift in the cellulose band because of the combined effect of high‐pressure processing and enzymatic hydrolysis. High‐pressure‐assisted enzymatic hydrolysis was determined the best combination to be ensured almost 18‐fold increase in SDF ratio and to be improved radical scavenging activity more than two times in addition to well‐documented change in viscoelastic properties of the GCRF. [ABSTRACT FROM AUTHOR]

The effects of alkaline pretreatment of walnut shells (WS) utilizing NaOH, Ca(OH)2, or KOH were studied. The highest lignin recovery was obtained from WS pretreated with NaOH‐glycerol. The optimum lignin recovery after pretreatment could be achieved when the NaOH/solid value was 20:1 (mL g−1). CaCl2 loading could enhance lignin flocculation, which improved the lignin recovery. Also, the efficiency of enzymatic hydrolysis was determined when xylanase and cellulase were added to the pretreated residue. WS pretreated in NaOH‐water and NaOH‐glycerol solutions were found to result in the highest xylose and glucose concentrations. This study indicates the broad application prospects of alkaline pretreatment of lignocellulosic biomass in agriculture and industry. [ABSTRACT FROM AUTHOR]

Postbiotics are defined as a "preparation of inanimate microorganisms and/or their components that confers a health benefit on the host." They can be produced by fermentation, using culture media with glucose (carbon source), and lactic acid bacteria of the genus Lactobacillus, and/or yeast, mainly Saccharomyces cerevisiae as fermentative microorganisms. Postbiotics comprise different metabolites, and have important biological properties (antioxidant, anti‐inflammatory, etc.), thus their cosmetic application should be considered. During this work, the postbiotics production was carried out by fermentation with sugarcane straw, as a source of carbon and phenolic compounds, and as a sustainable process to obtain bioactive extracts. For the production of postbiotics, a saccharification process was carried out with cellulase at 55°C for 24 h. Fermentation was performed sequentially after saccharification at 30°C, for 72 h, using S. cerevisiae. The cells‐free extract was characterized regarding its composition, antioxidant activity, and skincare potential. Its use was safe at concentrations below ~20 mg mL−1 (extract's dry weight in deionized water) for keratinocytes and ~ 7.5 mg mL−1 for fibroblasts. It showed antioxidant activity, with ABTS IC50 of 1.88 mg mL−1, and inhibited elastase and tyrosinase activities by 83.4% and 42.4%, respectively, at the maximum concentration tested (20 mg mL−1). In addition, it promoted the production of cytokeratin 14, and demonstrated anti‐inflammatory activity at a concentration of 10 mg mL−1. In the skin microbiota of human volunteers, the extract inhibited Cutibacterium acnes and the Malassezia genus. Shortly, postbiotics were successfully produced using sugarcane straw, and showed bioactive properties that potentiate their use in cosmetic/skincare products. [ABSTRACT FROM AUTHOR]

Among 24 isolated cellulolytic bacteria from Similipal Biosphere Reserve, the most efficient isolate was recognized as a strain of Bacillus albus. This strain of B. albus was evaluated for cellulase production and the cellulase activity was measured in submerged fermentation using substrate carboxymethyl cellulose (CMC). Different nutritional (carbon, nitrogen, and metal‐ion sources) and physical variables (pH, temperature, substrate concentration, and incubation time) during the growth of B. albus were optimized to obtain maximum cellulase activity. The highest cellulase activity of 5.79 U/mL for B. albus was observed at pH 6.75, temperature 37.5°C, CMC concentration 8.5 g/L, and 42 h incubation time. Further, supplementation of glucose as a subsidiary carbon source, yeast extract, peptone as nitrogen sources, and MgSO4 and MnSO4 as metal‐ion sources enhance the cellulase activity of B. albus. The purified enzyme was reported to have a molecular weight of ∼54 kDa as determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. A zymogram analysis evidenced the cellulase activity of the purified enzyme fractions obtained from diethylaminoethyl cellulose chromatography. The purified cellulase was reported to have an optimum pH and temperature of 7.0°C and 50°C, respectively with a capacity of retaining its 60% residual activity within pH 6.0–8.0 and temperature 30–40°C, respectively. The metal ions, K+ and Na+ were the activators, while Pb2+ and Hg2+ were the inhibitors for the purified cellulase. The purified cellulase showed Km and Vmax values of 0.38 M and 8.19 U/mL, respectively, in presence of the substrate CMC and also simultaneous consumption of both hexose and pentose sugars. [ABSTRACT FROM AUTHOR]

Sugarcane bagasse (SB) is a promising source of appreciable quantities of fermentable sugars. However, the presence of lignin hinders utilization of these carbohydrates and hence pretreatment to remove lignin is necessarily carried out. Here, a biological pretreatment method was synchronized with the production of a thermostable cellulase using SB as a raw material. Initially, bagasse was fermented by a laccase producing fungus, Trametes pubescens MB 89 under solid state fermentation (SSF) and a titer of 1758 IU mL−1 of laccase was obtained. Investigations of nine factors affecting laccase production through Plackett Burman design improved the titers to 6539 IU mL−1. Five factors (incubation period, concentration of CuSO4, temperature, moisture content, and particle size) were found significant which were optimized through Central Composite design leading to an improvement in the titers by ~5 folds (8841 IU mL−1). Biologically pretreated SB was fermented by a thermophilic bacterium, Neobacillus sedimentimangrovi UE25, that yielded 8.64 IU mL−1 of cellulase. Delignification and cellulose utilization were affirmed by structural analysis through FTIR and SEM. The synchronized process yielded higher titers of laccase and cellulase under SSF of SB with the minimum use of corrosive chemicals. [ABSTRACT FROM AUTHOR]

Cellulolytic actinobacterium, Promicromonospora sp. VP111 concomitantly produced cellulases (CELs), xylanase and pectinase when grown on commercial cellulose and untreated agricultural lignocellulosic residues (wheat straw and sugarcane bagasse). Secreted CELs hydrolyzed (enhanced with Co2+ ion) multiple cellulosic substrates, including sodium carboxymethyl cellulose (Na‐CMC), Whatman filter paper no. 1, microcrystalline cellulose (avicel), p‐nitrophenyl‐β‐D‐glucopyranoside (pNPG), laminarin, and cellulose powder. The CELs showed stabilities in the presence of various chemicals, including glucose (0.2 M), detergents (1%, w/v or v/v), denaturants (1%, w/v or v/v), and sodium chloride (NaCl, 30%, w/v). The CELs were fractionated using ammonium sulfate precipitation and dialysis. Activities (%) of fractionated CELs were retained at 60°C for endoglucanase/carboxymethyl cellulase (CMCase) (88.38), filter paper cellulase (FPase) (77.55), and β‐glucosidase (90.52), which indicated of thermo‐stability. Similarly, the activities (%) for CMCase (85.79), FPase (82.48), and β‐glucosidase (85.92) at pH 8.5 indicated of alkaline‐stability. Kinetic factors, Km and Vmax for endoglucanase component of fractionated CELs were 0.014 g/l and 158.23 µM glucose/min/mL, respectively. Fractionated CELs yielded activation energies (kJ/mol) of 17.933, 6.294, and 4.207 for CMCase, FPase, and β‐glucosidase activities, respectively in linear thermostable Arrhenius plots. Thus, this study reports on the multipurpose CELs from an untreated agricultural residue utilizing Promicromonospora in relation to broad substrate specificity, halo‐tolerance, alkaline‐tolerance, detergent‐tolerance, thermo‐tolerance, organic solvent‐tolerance, and end product‐tolerance. [ABSTRACT FROM AUTHOR]

Cellulases are used in textile, pulp and paper, brewery and wine, sugars, and ethanol industries. Four fungal isolates obtained from organic municipal solid wastes (OMSW) were selected based on their cellulolytic activity on carboxymethyl cellulose (CMC) agar medium. Based on the internal transcribed spacer (ITS) sequence of the ribosomal DNA, the four cellulolytic isolates were identified as Aspergillus fumigatus AKAL1, Aspergillus oryzae AKAL4, Aspergillus flavus AKAL8, and Aspergillus flavus AKAL9. After 9 days of fermentation at 30°C and pH 6.5 under 110 rpm agitation, these isolates produced the maximum amount of cellulase. The cellulase showed optimum activity at temperature 35–40°C and pH 6.0–7.0 and was stable for 1 h at 25–45°C and pH 5.0–7.0. The Mg2+ and Zn2+ significantly increased but Hg2+, K+, and Ca2+ severely repressed the cellulase activity. Degradation of filter papers and bio‐stoning of denim was successfully done with the crude cellulase. An endo‐β‐1,4‐glucanase was isolated and characterized from Aspergillus isolates. Genome‐wide analysis revealed that the genomes of A. oryzae, A. fumigatus, and A. flavus, the pertinent species of the fungal isolates, had 23, 25, and 22 cellulase genes, respectively. Phylogenetic analysis revealed that the cellulases in these fungal species were divided into three major groups, and the isolated endo‐β‐1,4‐glucanase clustered to Group II. Ten different motifs are present in cellulases of the three species. Results herein provide a valuable resource for understanding cellulase genes in Aspergillus species and potential application of cellulase in textile and fermentable sugars production industries. [ABSTRACT FROM AUTHOR]

In the process of bioethanol production, more stable and active cellulase in high temperature condition is required. In this study, syringic acid was applied in cellulase hydrolysis system. At 70°C, TvEG3 activity increased 201.36%, CtBglA activity decreased 72.79% by syringic acid. With syringic acid assisting, TvEG3 thermostability was improved, CtBglA thermostability was reduced. Syringic acid scarcely affected CtCBH. In hydrolysis system with the cellulases containing TvEG3, CtCBH, and CtBglA, the reducing sugar yield improved by 28.37% with syringic acid assisting. With the molecular dynamic simulation in syringic acid system, the backbone root‐mean‐square deviation (RMSD) and the residue root‐mean‐square fluctuation (RMSF) of TvEG3, CtCBH reduced, while the RMSD and RMSF of CtBglA increased. The reduction in the number of secondary structures, especially α‐helix, caused the structure of CtBglA in the presence of syringic acid to collapse at high temperature. More secondary structures in TvEG3 and more α‐helix in CtCBH in the presence of syringic acid make them more stable at high temperatures. These means syringic acid can stabilize TvEG3 and CtCBH structure, destabilize CtBglA structure at high temperature. In summary, this study not only provides insight into cellulase hydrolysis at high temperature with syringic acid assisting but also demonstrates the promoting mechanism of syringic acid. [ABSTRACT FROM AUTHOR]

Lignocellulosic residues are amongst the most abundant waste products on Earth. Therefore, there is an increasing interest in the utilization of these residues for bioethanol production and for biorefineries to produce compounds of industrial interest. Enzymes that breakdown cellulose and hemicellulose into oligomers and monosaccharides are required in these processes and cellulolytic enzymes with optimum activity at a low pH area are desirable for industrial processes. Here, we explore the fungal biodiversity of Rıo Tinto, the largest acidic ecosystem on Earth, as far as the secretion of cellulolytic enzymes is concerned. Using colorimetric and industrial substrates, we show that a high proportion of the fungi present in this extremophilic environment secrete a wide range of enzymes that are able to hydrolyze cellulose and hemicellulose at acidic pH (4.5–5). Shotgun proteomic analysis of the secretomes of some of these fungi has identified different cellulases and hemicellulolytic enzymes as well as a number of auxiliary enzymes. Supplementation of pre‐industrial cocktails from Myceliophtora with Rio Tinto secretomes increased the amount of monosaccharides released from corn stover or sugar cane straw. We conclude that the Rio Tinto fungi display a good variety of hydrolytic enzymes with high industrial potential. [ABSTRACT FROM AUTHOR]

Beach wrack constitutes an unutilized, abundant renewable source of marine macro algae being rich in valuable biopolymers. This work investigates the revalorization of the brown algae Fucus vesiculosus collected from beach wrack as a source of alginate, the main polysaccharide in brown algae which constitutes a potential ingredient for the development of biopolymer films. Enzyme‐assisted extraction has been investigated for cell disruption with commercial cellulase blends and proteases. The effect of enzyme type, enzyme activity, and extraction time on alginate yields, molecular weight distribution, functional groups, and purity has been studied. [ABSTRACT FROM AUTHOR]

Fungi play a critical role in the deterioration of ancient papyri due to their organic and hygroscopic nature. Fourteen fungal species belonging to five genera—Aspergillus, Cladosporium, Penicillium, Talaromyces, and Ulocladium—were isolated and identified from 20 Egyptian papyrus artifacts located at the Grand Egyptian Museum—Conservation Center (GEM‐CC). The biodeterioration mechanism was evaluated by two analyses: cellulase enzyme activity and acid production assay. The isolated fungus P. lanosum showed potent biodeterioration power as it exhibited the highest enzymatic activity and could produce acid as well. Four different analyses were performed to assess the entity of the biodeterioration phenomenon: scanning electron microscopy, Fourier transform infrared spectroscopy, colorimetry, and pH measurements. The analyses demonstrated considerable changes in surface morphology, chemical structure (mainly represented by oxidation and hydrolysis of infected papyrus samples), darkness of the papyrus samples (ΔE 3.84–7.33), and pH (which dropped to 5.2 in the case of A. niger‐infected sample. This article builds a preliminary perception for monitoring the biodeterioration of archaeological papyri and a trial for in vitro treatment to prevent further degradation, which may lead to loss of our valuable artifacts over a prolonged period. [ABSTRACT FROM AUTHOR]

Fungi are of great importance in biotechnology, for example in the production of enzymes and metabolites. The main goal of this study was to obtain a high‐coverage draft of the Stachybotrys microspora genome and to annotate and analyze the genome sequence data. The rare fungus S. microspora N1 strain is distinguished by its ability to grow in an alkaline halophilic environment and to efficiently secrete cellulolytic enzymes. Here we report the draft genome sequence composed of 3715 contigs, a genome size of 35 343 854 bp, with a GC content of 53.31% and a coverage around 20.5×. The identification of cellulolytic genes and of their corresponding functions was carried out through analysis and annotation of the whole genome sequence. Forty‐six cellulases were identified using the fungicompanion bioinformatic tool. Interestingly, an S. microspora endoglucanase selected from those with a low isoelectric point was predicted to have a halophilic profile and share significant homology with a well‐known bacterial halophilic cellulase. These results confirm previous biochemical studies revealing a halophilic character, which is a very rare feature among fungal cellulases. All these properties suggest that cellulases of S. microspora may have potential for use in the biofuel, textile, and detergent industries. [ABSTRACT FROM AUTHOR]

4‐hydroxybenzoic acid (4‐HBA) is an industrially important aromatic compound, and there is an urgent need to establish a bioprocess to produce this compound in a sustainable and environmentally friendly manner from renewable feedstocks such as cellulosic biomass. Here, we developed a bioprocess to directly produce 4‐HBA from cellulose using a recombinant Pichia pastoris strain that displays heterologous cellulolytic enzymes on its cell surface via the glycosylphosphatidylinositol (GPI)‐anchoring system. β‐glucosidase (BGL) from Aspergillus aculeatus, endoglucanase (EG) from Trichoderma reesei, and cellobiohydrolase (CBH) from Talaromyces emersonii were co‐displayed on the cell surface of P. pastoris using an appropriate GPI‐anchoring domain for each enzyme. The cell‐surface cellulase activity was further enhanced using P. pastoris SPI1 promoter‐ and secretion signal sequences. The resulting strains efficiently hydrolyzed phosphoric acid swollen cellulose (PASC) to glucose. Then, we expressed a highly 4‐HBA‐resistant chorismate pyruvate‐lyase (UbiC) from Providencia rustigianii in the cellulase‐displaying strain. This strain produced 975 mg/L of 4‐HBA from PASC, which corresponding to 36.8% of the theoretical maximum yield, after 96 h of batch fermentation without the addition of commercial cellulase. This 4‐HBA yield was over two times higher than that obtained from glucose (12.3% of the theoretical maximum yield). To our knowledge, this is the first report on the direct production of an aromatic compound from cellulose using cellulase‐displaying yeast. [ABSTRACT FROM AUTHOR]

Aqueous enzymatic extraction (AEE) of oil from Acer truncatum Bunge seed kernel was investigated. The effects of enzyme type on the extraction yield of oil were studied, and the results showed that the oil yield obtained with pentosanase was higher than that obtained with the other enzymes. The combination of pentosanase and cellulase showed better extraction performance than a single enzyme, and the operation parameters of the AEE method were optimized. A maximum oil yield of 37.94% was obtained. The analysis results of chemical compositions of the extracted oils showed that the content of unsaturated fatty acids in the oil extracted by the AEE method was 90.28%, and the content of nervonic acid was about 5.59%. In addition, the main physical and chemical properties of A. truncatum Bunge seed oil were measured. The oil obtained by the AEE method met the China National Standard of A. truncatum Bunge oil. [ABSTRACT FROM AUTHOR]

Obtaining a robust and applicable enzyme for bioethanol production is a dream for biorefinery engineers. Herein, we describe a general method to evolve an all‐round and interpretable enzyme that can be directly employed in the bioethanol industry. By integrating the transferable protein evolution strategy InSiReP 2.0 (In Silico guided Recombination Process), enzymatic characterization for actual production, and computational molecular understanding, the model cellulase PvCel5A (endoglucanase II Cel5A from Penicillium verruculosum) was successfully evolved to overcome the remaining challenges of low ethanol and temperature tolerance, which primarily limited biomass transformation and bioethanol yield. Remarkably, application of the PvCel5A variants in both first‐ and second‐generation bioethanol production processes (i. Conventional corn ethanol fermentation combined with the in situ pretreatment process; ii. cellulosic ethanol fermentation process) resulted in a 5.7–10.1 % increase in the ethanol yield, which was unlikely to be achieved by other optimization techniques. [ABSTRACT FROM AUTHOR]

This study shows the morphological identification of anaerobic fungal strains isolated from fecal samples of goats inhabiting Turkey and the effects of various metal ions and chemicals on extracellular xylanase production. Three different anaerobic gut fungi isolated from wild goats in Turkey were identified as Neocallimastix spp. xylanase, cellulase, and lichenase production were tested in culture supernatants, and the maximum‐specific activities were found as 560.42 ± 9.39, 159.70 ± 3.88, and 157.36 ± 3.83 (μmol/min/mg protein), respectively. While the optimum temperature range of exo‐xylanases was found as 40–50°C, their optimum pH range was determined as 6.0–6.5. Xylanase activity decreased in metal ions and other chemical reactants based on dose. The metal ion that significantly inhibited xylanase activity was Fe+3. It was found that the ferric ions inhibited xylanase activity in all three anaerobic gut fungi by 30%–90% depending on molarity. On the contrary, the 1 mM concentrations of the Mn+2, Ba+2, Co+2, Cu+2, Sn+2, and Mg+2 metal ions and the ethylenediaminetetraacetic acid and β‐mercaptoethanol reagents had a positive effect at rates in the range of 3%–92%. In conclusion, these findings demonstrate that anaerobic gut fungus has very stable fibrolytic enzymes that need to be separated, as well and the existence of a unique resource for industrial applications. [ABSTRACT FROM AUTHOR]

Vermicomposting involves enrichment of microorganisms that are able to resist higher temperatures and perform simultaneous degradation of lignocellulose, and therefore, such microbial communities are a potential source of cellulolytic enzymes. This study aimed to optimize the production of a processive cellulase by Parageobacillus thermoglucosidasius NBCB1 isolated from vermicompost, under submerged fermentation of rice straw and to characterize the purified enzyme for industrial suitability. Cellulase production in basal medium (7.27 IU/mg) was enhanced to 61 IU/mg by One Factor At a Time approach, which was further improved to 78.46 IU/mg by genetic algorithm based artificial neural networking. The cellulase PtCel1 purified from bacterial culture showed a molecular weight of ≈33 kD, had activity on both crystalline (305 IU/mg) and amorphous (184 IU/mg) cellulose as substrates. It had pH and temperature optima of 5.5°C and 60°C, respectively, and retained 100% activity upon preincubation at 60°C for 1 h indicating thermostability. PtCel1 was tolerant to sodium dodecyl sulfate, glucose and mannose; and the various metal chlorides, such as sodium, magnesium, calcium and zinc, acted as inducers giving 77.54%, 45.15%, 61.10%, and 169.14% augmentation of activity, respectively. Its efficiency on cellulosic substrates and robustness against aforementioned chemical and thermal environment makes it suitable for industrial applications. [ABSTRACT FROM AUTHOR]

The processive cellulase (CelO) is an important modular enzyme of Clostridium thermocellum. To study the effect of the carbohydrate‐binding module (CBM3b) on the catalytic domain of CelO (GH5), four engineered derivatives of CelO were designed by truncation and terminal fusion of CBM3b. These are CBM at the N‐terminus, native form (CelO‐BC, 62 kDa); catalytic domain only (CelO‐C, 42 kDa); CBM at the C‐terminus (CelO‐CB, 54 kDa) and CBM attached at both termini (CelO‐BCB, 73 kDa). All constructs were cloned into pET22b (+) and expressed in Escherichia coli BL21 (DE3) star. The expression levels of CelO‐C, CelO‐CB, CelO‐BC, and CelO‐BCB were 35%, 35%, 30%, and 20%, respectively. The enzyme activities of CelO‐C, CelO‐CB, CelO‐BC, and CelO‐BCB against 1% regenerated amorphous cellulose (RAC) were 860, 758, 985, and 1208 units per μmole of the enzyme, respectively. The enzymes were partially purified from the lysate of E. coli cells by heat treatment followed by anion exchange FPLC purification. Against RAC, CelO‐C, CelO‐CB, CelO‐BC, and CelO‐BCB showed KM values of 32, 33, 45, and 43 mg⋅mL–1 and Vmax values of 3571, 3846, 3571, and 4545 U⋅min–1, respectively. CBM3b at the N‐terminus of GH5 linked through a P/T‐rich linker was found to enhance the catalytic activity and thermostability of the enzyme. [ABSTRACT FROM AUTHOR]