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The mulberry branch can be used as a renewable biomass resource for the production of biofuels since it contains a significant amount of lignocellulose. However, the rapid deactivation of enzymes during lignocellulose degradation in a high-temperature environment leads to high costs and low efficiency of enzymatic saccharification. In this study, xylanase was modified by rational design, and xylanase and cellulase were used to synergistically degrade mulberry branches pretreated with seawater. The fermentable sugar yield and degree of synergy (DS) were determined. The dominant mutant S21Y/N318W of Hortaea werneckii xylanase Hwxyl10A was obtained with improved thermostability (T 50 increased by 12 °C, t 1/2 is 17-fold that of WT). Meanwhile, the specific activity at 75 °C increased from 1470 to 1901 U/mg. Under the condition of seawater immersion pretreatment, the highest yield of fermentable sugar was 313.5 μmol/g, and the highest DS was 1.57 after enzymatic hydrolysis of mulberry branches by cellulase and S21Y/N318W at 50 °C. Therefore, the biotransformation of mulberry branches reducing sugar has been significantly improved, which provides an efficient method of biomass saccharification for biofuel production. [Display omitted] • An efficient xylanase with improved properties was obtained by rational design technology. • The thermal stability of mutants were improved by enhancing the rigidity of flexible region. • The specific activity at 75 °C of xylanase mutant S21Y/N318W was increased by 29%. • The half-life of xylanase mutant S21Y/N318W was prolonged by 17 times. • The degradation was strengthened by the synergistic catalysis of S21Y/N318W and cellulase. [ABSTRACT FROM AUTHOR]

• As the angle between the blastholes widens, the fractal dimension of the crushing zone and blasting crack among the blast borehole decreases accordingly, suppressed the blasting effect of the blasted medium. • The von Mises strain-time curve of the specimen initially increases and then decreases. As the angle between the blastholes widens, the peak strain diminishes, and the time to reach peak strain lengthens. • On the central axis between the two holes, the stress superposition becomes more obvious in the direction away from the initiation point, and the peak value of strain is the highest at the bottom of the blastholes. • When the angle between the blastholes is 30°, the strain in the specimen takes the longest to decline from peak to valley, thus maximizing the use of blasting energy. The drilling and blasting method is extensively employed in various industries such as mining and infrastructure construction. In the majority of blasting operations, strip-charges are predominantly utilized, while blastholes often exhibit an angled orientation in practical engineering scenarios. To investigate the stress wave superposition effect and the crack propagation law among blastholes at varying angles, this study employs a high-speed digital image correlation system to conduct blasting model experiments. The results indicate that: the fractal dimension of the crack propagation trajectory decreases progressively as the angle increases. the von Mises strain-time curve of the specimen initially increases and then decreases. As the angle between the blastholes widens, the fractal dimension of the crushing zone and blasting crack among the blast borehole decreases accordingly, suppressed the blasting effect of the blasted medium. On the central axis between the two holes, the stress superposition becomes more obvious in the direction away from the initiation point, and the peak value of strain is the highest at the bottom of the blastholes. When the angle between the blastholes is 30°, the strain in the specimen takes the longest to decline from peak to valley, and the blasting stress wave sustains its influence for an extended duration, thus maximizing the use of blasting energy. Conversely, when the angle between the blastholes is 0°, the specimen experiences the shortest duration of blasting action and the least efficient energy utilization. [ABSTRACT FROM AUTHOR]

Most laparoscopic surgeries under general anesthesia are performed in noisy environments, although the effect of intraoperative noise reduction on postoperative pain remains uncertain. This study aimed to explore whether postoperative pain could be reduced through the intraoperative use of noise-cancelling headphones. This study was conducted as a prospective parallel-group randomized clinical trial. Operating room and surgery room. Ninety patients who underwent laparoscopic surgery under general anesthesia. In the intervention group, noise-cancelling headphones were used to reduce noise intensity during laparoscopic surgery under general anesthesia. The primary outcome was the maximum movement-evoked pain intensity within 24 h post-surgery, measured using a 10-point numeric rating scale. Secondary outcomes included the maximum resting pain score and total opioid consumption during the 24-h period post-surgery. Mean intraoperative noise and the proportion of intraoperative time with noise intensity ≥70 dB were recorded. The maximum movement-evoked pain score was significantly lower in the intervention group than in the control group (mean score [SD], 2.7 [1.0] and 4.0 [1.0], respectively; P < 0.001). The intervention group required significantly fewer opioids than the control group (mean [SD], 44.2 [12.8] and 51.3 [17.5] mg, respectively; P = 0.032). In the control group, but not the intervention group, all postoperative pain scores were significantly associated with the proportion of intraoperative time with noise intensity ≥70 dB, which was an independent risk factor for postoperative pain. During laparoscopic surgery under general anesthesia, intraoperative noise isolation using noise-cancelling headphones is a safe and effective strategy for relieving postoperative pain and decreasing total opioid analgesic consumption. [Display omitted] • Noise-cancelling headphones during surgery reduced postoperative pain scores. • Noise-cancelling headphones during surgery reduced postoperative opioid consumption. • Reduction of hyperpathia contributes to the beneficial effect of noise-cancelling. [ABSTRACT FROM AUTHOR]

Xylanase is widely used for the degradation lignocellulosic biomass as an important accessory enzyme. But productive hydrolysis of the recalcitrant lignocellulose also requires for applicable pretreatment with environmental friendliness. Meanwhile, the complicated industrial processes also demand for catalytically efficient enzymes with more resistance to both pH and heating conditions, which barely exist for the wild types. In this study, the dominant mutant H51K/Q118A from Gloeophyllum trabeum derived GtXyn10 (WT) was obtained by directed evolution. Compared to the WT, apart from the impressive pH resistance from pH 1.0–9.0, the dominant mutant H51K/Q118A also exhibited higher specific activity (2.5-fold; 2510 vs. 1010 U/mg), higher catalytic efficiency (2.4-fold; 460 vs. 190 mL/s‧mg), and better thermostability (with t 1/2 at 80 °C extend by 10 min). When 'Seawater + Feton' pretreatment was applied, the lignin clearance rate reached 62.5%, 86.6% higher than that of feton pretreatment (33.5%). After pretreatment of 'Seawater + Feton', Compared with the cellulase-only treatment (159 μmol/g), synergistic hydrolysis with both H51K/Q118A and cellulase (236 μmol/g) increased the fermentable sugar yields from bagasse by 48.4%. In combination with the optimized pretreatment and synergistic hydrolysis of the modified xylanase and cellulase, the fermentable sugar production for green bio-energy was realized with efficiency. [Display omitted] • An efficient xylanase with improved properties was obtained by directed evolution. • Steric hindrance elimination was a possible mechanism for catalytic enhancement. • 'Seawater + Feton' pretreatment promoted the removal of lignin in bagasse. • Synergistic hydrolysis (H51K/Q118A + cellulase) realized the optimal degradation. • Biomass utilization for fermentable sugars was optimized by combined strategies. [ABSTRACT FROM AUTHOR]

Abstract In this paper, a novel one-step solution approach is developed to prepare high quality methylammonium lead trihalide (MAPbI 3) perovskite films. This approach employs methanol as additive, which is added into the perovskite precursor solution. The use of methanol can enhance the absorption of MAPbI 3 and significantly improve the coverage of MAPbI 3 on a planar substrate. The fabricated perovskite solar cell (PSC) with methanol exhibits a higher power conversion efficiency (PCE) compared with that without methanol (19.51% vs 16.53%). Furthermore, the power conversion efficiencies of devices with methanol are still high after kept in dark under ambient environment for 30 days. This work provides a method to prepare high quality perovskite films for planar perovskite solar cells with high performance. Graphical abstract A novel one-step solution is reported to prepare perovskite films by adding methanol into the perovskite precursor solution. The use of methanol can enhance the absorption of CH 3 NH 3 PbI 3 and significantly improve the coverage of CH 3 NH 3 PbI 3 on a planar substrate. The fabricated perovskite solar cell with methanol exhibits a higher power conversion efficiency compared with that without methanol (19.51% vs 16.53%). Image 1 Highlights • Methanol was employed as additive successfully for perovskite solar cells. • The methanol-treated PSCs showed remarkable efficiency and enhanced stability. • The methanol-treated method can suppress the hysteresis. [ABSTRACT FROM AUTHOR]

The vast coastline provides Canada with a flourishing seafood industry including bivalve shellfish production. To sustain a healthy bivalve molluscan shellfish production, the Canadian Shellfish Sanitation Program was established to monitor the health of shellfish harvesting habitats, and fecal coliform bacteria data have been collected at nearly 15,000 marine sample sites across six coastal provinces in Canada since 1979. We applied Functional Principal Component Analysis and subsequent correlation analyses to find annual variation patterns of bacteria levels at sites in each province. The overall magnitude and the seasonality of fecal contamination were modelled by functional principal component one and two, respectively. The amplitude was related to human and warm-blooded animal activities; the seasonality was strongly correlated with river discharge driven by precipitation and snow melt in British Columbia, but such correlation in provinces along the Atlantic coast could not be properly evaluated due to lack of data during winter. • Sparse fecal pollution data were collected across ~15,000 sites along Canada's coast. • Longitudinal geospatial distributions of bacterial levels over decades were studied. • Functional Principal Component Analysis revealed two major annual patterns. • Human and wild-life activities related to the geospatial bacterial amplitudes. • River discharge and precipitation related to the temporal changes of fecal bacteria. [ABSTRACT FROM AUTHOR]

• The pre-crack initiates anew when the horizontal distance between the moving crack and the semicircular arc pre-crack is small. When the distance increases to 8.33 mm, the crack no longer penetrates the semicircular arc pre-crack, and the pre-crack attracts the crack first and then repels it. When d = 11.67 mm, the pre-crack has no considerable effect on the moving crack. • The crack deflection angle and the horizontal offset distance of the two groups of experiments conform to a linear function. • Setting the semicircular arc pre-crack outside the protected medium can effectively protect the medium from impact load. When the moving crack is outside the arc, the medium inside the arc pre-cracks are in the stress relief zone, which can be well protected. In this paper, a plexiglass (PMMA) specimen is designed for dynamic impact testing based on the digital laser dynamic caustics experimental system. The impact of offset distance on dynamic fracture behavior of PMMA specimens with semicircular arc pre-cracks in two different directions is investigated. Research shows that: the pre-crack initiates anew when the horizontal distance between the moving crack and the semicircular arc pre-crack is small. With the increase of horizontal distance, the pre-crack attracts the crack first and then repels it. When the distance reaches a critical threshold, the pre-crack has no considerable effect on the moving crack. The above conclusions are not affected by the direction of the semicircular arc pre-crack. Furthermore, the crack deflection angle and the horizontal offset distance of the two groups of experiments conform to a linear function. When the horizontal offset distance is small, the middle part of the pre-crack initiates again. When the moving crack is outside the arc pre-crack, the medium in the pre-crack is the stress relief zone, according to the numerical data. The ability of the medium to resist impact damage can be effectively boosted by placing acceptable pre-cracks on the medium's periphery. [ABSTRACT FROM AUTHOR]

[Display omitted] • Mutant from XYL10C_ΔN with improved catalytic activity was obtained through protein engineering. • Synergistic hydrolysis with the engineered xylanase and cellulase excelled other enzyme treatments. • Seawater immersion was helpful for biomass hydrolysis as chemical pretreatment. • Ramulus mori was converted into fermentable sugars with lower energy consumption. Biorefinery of Ramulus mori with lower energy consumption through improved enzyme and pretreatment strategies was reported. Directed evolution and saturation mutagenesis were used for the modification of xylanase, the yield of fermentable sugars and the degree of synergy (DS) were determined for different pretreatment (seawater/non-seawater) and enzyme treatment groups (xylanase/cellulase/co-treatment). The dominant mutant I133A/Q143Y of Bispora sp. xylanase XYL10C_ΔN was obtained with improved specific activity (1860 U/mg), catalytic efficiency (1150 mL/s∙mg) at 40 °C, and thermostability (T 50 increased by 7 °C). With the pretreatment of seawater immersion, the highest yield of fermentable sugars for Ramulus mori at 40 °C reached 199 μmol/g when hydrolyzed with cellulase and I133A/Q143Y, with the highest DS of 2.6; this was 4.5-fold that of the group hydrolyzed by cellulase alone with non-seawater pretreatment. Thus, bioconversion of reducing sugar from Ramulus mori was improved significantly at lower temperatures, which provides an efficient and energy-saving way for biofuel production. [ABSTRACT FROM AUTHOR]

[Display omitted] • An efficient bagasse biorefinery with low energy consumption was realized. • A thermostable xylanase mutant was engineered for catalysis under mild conditions. • Seawater following steam explosion promotes the enzymatic hydrolyzation of bagasse. • Synergistic hydrolysis of high activity xylanase and cellulase was optimal. Lignocellulosic biorefining for producing biofuels poses technical challenges. It is usually conducted over a long time using heat, making it energy intensive. In this study, we lowered the energy consumption of this process through an optimized enzyme and pretreatment strategy. First, the dominant mutant M137E/N269G of Bispora sp. MEY-1 XYL10C_ΔN was obtained by directed evolution with high catalytic efficiency (970 mL/s∙mg) and specific activity (2090 U/mg) at 37 °C, and thermostability was improved (T 50 increased by 5 °C). After pretreatment with seawater immersion following steam explosion, bagasse was co-treated with cellulase and M137E/N269G under mild conditions (37 °C), the resulting highest yield of fermentable sugars reached 219 µmol/g of bagasse, 46% higher than that of the non-seawater treatment group, with the highest degree of synergy of 2.0. Pretreatment with seawater following steam explosion and synergistic hydrolysis through high activity xylanase and cellulase helped to achieve low energy degradation of lignocellulosic biomass. [ABSTRACT FROM AUTHOR]

Developing circular economy is the only way to improve the efficiency of resource utilization. Whole-cell catalysis is an effective method to recycle enzymes, improve catalytic efficiency, and reduce production costs. The enzyme, α -L-rhamnosidase has considerable application prospects in the field of biocatalysis as it can hydrolyze a variety of α -L rhamnoses. In the present study, the genes for α -L-rhamnosidase (rhaB1) and enhanced fluorescent protein (EGFP) were co-expressed using a bi-promoter expression vector pRSFDuet1 and their enzymatic properties were evaluated. To our knowledge, this study has established an effective rhamnosidase-fluorescent indicator and whole-cell catalytic system for the first time. Moreover, we analyzed the change in the activity of the crude rhaB1-EGFP as well as its whole-cell during the biocatalysis process using fluorescence intensity. Recombinant rhaB1-EGFP as a product which contains rhaB1 and EGFP showed higher thermal stability, pH stability, and conversion efficiency than rhaB1, and its optimum temperature for rutin catalysis was ideal for industrial applications. Moreover, under the optimal conditions of a rutin concentration of 0.05 g/L, pH of 6.0, temperature of 40 °C, a yield of 92.5% was obtained. Furthermore, we demonstrated the relationship between the fluorescence intensity and enzyme activity. This study established a highly efficient whole-cell catalytic system whose activity can be evaluated by fluorescence intensity, providing a reference for enzyme recycling. Unlabelled Image • Two genes of rhaB1 and EGFP were firstly co-expressed by a bi-promoter expression vector. • Crude rhaB1-EGFP possessed higher thermal and pH stabilities than those of crude rhaB1. • Expression vector activity could be quantitatively detected by measuring fluorescence intensity. • Affinity of rhaB1-EGFP to substrate was not significantly reduced confirmed by SPR assay. • A labeled and recycling whole-cell catalyst was developed for flavonoids transformation. [ABSTRACT FROM AUTHOR]

The pupae is one of the most valuable components of the silkworm, a by-product of silk reeling. Employing them as a raw material to create valued products will help to recycle and reuse silkworm pupae, as well as contribute to the food industry's long-term development. Silkworm pupae protein (SPP) and pupae oil (SPO) are high in nutritional and bioactive components that have a variety of applications in the food industry as well as health-promoting characteristics. In this review, we first elaborate on the modification, nutritional, and functional properties of silkworm pupae in terms of requirements for protein and lipids processing as well as current extraction technology. In the subsequent sections, we summarize the modification techniques including the science and technology involved in attaining fatty acid components of SPO. Finally, we focus on recent studies related to the utilization of SPP and SPO subject to their inclusion in foods for their health benefits. The key to silkworm pupae product design may be to fully exploit and utilize the inherent characteristics of the pupae to produce functional peptides and structural lipids with high nutritional content, which may necessitate further improvements in the bioactive and functional properties. Designing more ecologically friendly, low-cost, and efficient processes for producing protein and oil from silkworm pupae is also a trend in technology. This may further provide new insights into the mechanisms behind modifying silkworm pupae as food and its health advantages. [Display omitted] • Modifications and functions of silkworm pupae protein (SPP) and oil (SPO) were reviewed. • Modification of SPP and SPO reveal valuable nutritional properties. • Inclusion of SPP and SPO in food and feed applications were summarized. • Peptide composition in silkworm pupae aid in valuable biological activities. • Further research to compare SPP and SPO with standard proteins and oils is recommended. [ABSTRACT FROM AUTHOR]

[Display omitted] • A novel scheme for efficient conversion of flavonoid glycosides by biofilms was developed. • Gene regulation and carrier modification are promising strategies to enhance biofilm catalysis. • The biomass of biofilms from csg D overexpressed strain significantly increased by 79.35 %. • The biofilms adhesion of E. coli in strong oxidant modified carrier was increased by 105.80 %. • The catalytic efficiency of E. coli biofilms enhanced bi-directional was increased by 247.85 %. The biofilm of an engineered strain is limited by slow growth and low yield, resulting in an unsatisfactory ability to resist external stress and promote catalytic efficiency. Here, biofilms used as robust living catalysts were manipulated through dual functionalized gene regulation and carrier modification strategies. The results showed that gene overexpression regulates the autoinducer-2 activity, extracellular polymeric substance content and colony behavior of Escherichia coli , and the biofilm yield of csg D overexpressed strains increased by 79.35 % compared to that of the wild type strains (p < 0.05). In addition, the hydrophilicity of polyurethane fibres modified with potassium dichromate increased significantly, and biofilm adhesion increased by 105.80 %. Finally, the isoquercitrin yield in the catalytic reaction of the biofilm reinforced by the csg D overexpression strain and the modified carrier was 247.85 % higher than that of the untreated group. Overall, this study has developed engineered strains biofilm with special functions, providing possibilities for catalytic applications. [ABSTRACT FROM AUTHOR]

TEC induced on the top of PCBM have been studied as the cathode-interfacial layer (CIL) and the CIL/perovskite interface is modified with a well-matched energy band alignment and suppressed defects. Enhanced power conversion efficiency with negligible hysteresis is obtained for planar perovskite solar cells. • A seamless PCBM@TEC capped perovskite film for the inverted planar PSCs. • PCBM@TEC CIL induced obvious efficiency enhancement for inverted PSCs. • Non-encapsulated devices based on the hydrophobic PCBM@TEC CIL improved stability of planar PSCs. • These improvements in performance and stability highlight the merit of using PCBM@TEC as interfacial layer in planar PSCs. Among the factors that lead to the reduction of the efficiency and stability of perovskite solar cells (PSCs) using [6,6]-phenyl C61 butyric acid methyl ester (PCBM) versus those of conventional structure, is the difficulty involved in realizing a high-quality film, and the non-radiative recombination that takes place at the interface between PCBM and perovskite. Our present work proposes a fabrication technique capable of overcoming these issues. Organic small molecule material was introduced along with PCBM, to obtain a homogeneously seamless film that plays the role of a dipole layer. The incorporated triethyl citrate (TEC) boosts the power conversion efficiency (PCE) from 14.56% to 17.86% with suppressed hysteresis, which originates primarily from the efficient electron transfer between the perovskite/PCBM and the Ag cathode interface. The results of electrochemical impedance spectroscopy measurements imply efficient electron transfer, low series resistance, and large recombination resistance in the photovoltaic device employing PCBM@TEC as the cathode interfacial layer. Specifically, the highly hydrophobic PCBM@TEC cathode interfacial layer (CIL) showed higher resistance to moisture, leading to a more environmentally-stable device. The results also showed the efficiency of the device retained 84% of its initial value after 30 days of operation under ambient conditions, which shows a visibly superior stability compared to a reference device that, under the same environment, retained only 37% its initial efficiency. Such advancement in the operational efficiency and stability of the device, will contribute to the longevity and cost-effectiveness of inverted PSC devices. [ABSTRACT FROM AUTHOR]

Efficient hydrolysis of pectic materials to sugars requires the synergistic action of endo- and exo-polygalacturonases. Two novel polygalacturonases (exo- Te PG28a and endo- Te PG28b) were identified in Talaromyces leycettanus JCM12802, overexpressed in Pichia pastoris , and characterized in this report. The specific activities of Te PG28a and Te PG28b towards polygalacturonic acid were 280 ± 9 and 25,900 ± 502 U/mg, respectively. Both enzymes exhibited optimal activities at pH 3.5 and retained highly stable over a broad pH range of 2.0–7.0. Distinct from most fungal polygalacturonases that have low temperature optima, Te PG28a and Te PG28b were optimally active at 70 °C. When treated the grape juice with the enzyme combination (the unit ratio of Te PG28a: Te PG28b was 1:4), higher pectin-degrading efficiency (up to 140%) was achieved, and light transmittance was improved from 14% to 82%. These favorable enzymatic properties make Te PG28a and Te PG28b attractive for the applications in the juice industry. [ABSTRACT FROM AUTHOR]

β-1,3–1,4-Glucanase is an indispensable biocatalyst in barley brewing industry for its crucial effect in reducing the viscosity of mash. However, the unsatisfactory thermostability greatly limited its application performance. In this study, structure-based surface charge engineering was conducted aiming at thermostability improvement of BisGlu16B, a highly active β-1,3–1,4-glucanase from Bispora sp. MEY-1. By applying the enzyme thermal stability system (ETSS), residues D47, D213, and D253 were inferred to be critical sites for thermal properties. Single (D47A, D213A, and D253A) and combination (D47A/D213A/D253A) mutants were generated and compared with BisGlu16B. Among all improved mutants, D47A/D213A/D253A outstanded in thermostability. In comparison with BisGlu16B, its T 50 and T m were respectively increased by 7.0 °C and 4.3 °C, while the t 1/2 at 70 °C was 8.1 times that of the wild type. Furthermore, the catalytic activity of D47A/D213A/D253A also increased by 42.5%, compared with BisGlu16B (42,900 ± 300 U/mg vs. 30,100 ± 800 U/mg). Comparing with BisGlu16B and commercial enzyme treatment groups, under simulated malting conditions, efficiency improvement was observed in decreasement of viscosity (35.5% and 90.7%) and filtration time (30.9% and 34.6%) for D47A/D213A/D253A treatment group. Molecular dynamics simulation showed that mutation sites A47, A213, and A253 increased the protein rigidity by lowering the overall root mean square deviation (RMSD). This study may bring optimization of technology and improvement of producing efficiency to the present brewing industry. [Display omitted] • Surface charge engineering protects the catalytic core during modification. • The β-glucanase activity was improved besides the intended higher thermostability. • Reduced mash viscosity and filtration time was observed during barley malting. • The study provided a more efficient catalyst for brewing industry. [ABSTRACT FROM AUTHOR]

A xylanase gene of GH 10, Tlxyn10A , was cloned from Talaromyces leycettanus JCM12802 and expressed in Pichia pastoris . Purified recombinant Tl Xyn10A was acidic and hyperthermophilic, and retained stable over the pH range of 2.0–6.0 and at 90 °C. Sequence analysis of Tl Xyn10A identified seven residues probably involved in substrate contacting. Three mutants ( Tl Xyn10A_P, _N and _C) were then constructed by substituting some or all of the residues with corresponding ones of hyperthermal Xyl10C from Bispora sp. MEY-1. Tl Xyn10A_P with mutations at subsites +2 to +4 exhibited improved specific activity (by 0.44-fold) and pH stability (2.0–10.0). Molecular dynamics simulation analysis indicated that mutations E229I and F232E probably weaken the substrate affinity at subsites +3 to +4, and G149D may introduce a new hydrogen bond. These modifications altogether account for the improved performance of Tl Xyn10A_P. Moreover, Tl Xyn10A_P was able to hydrolyze wheat straw persistently, and has the application potentials in various industries. [ABSTRACT FROM AUTHOR]

A microwave-assisted and phosphine-mediated Tomita Zipper cyclization of dicyanomethylideneoxindoles and ynones has been developed. Various functionalized spirooxindoles with five-membered carbocyclic ring can be obtained in moderate to good yields with moderate to excellent diastereoselectivities through in situ generation of α-nucleophile (up to 87% yield, >20:1 dr). [ABSTRACT FROM AUTHOR]

A xylanase gene of glycoside hydrolase family 10, Gt Xyn10, was cloned from Gloeophyllum trabeum CBS 900.73 and expressed in Pichia pastoris GS115. Purified recombinant Gt Xyn10 exhibited significant activities to xylan (100.0%), lichenan (11.2%), glucan (15.2%) and p -nitrophenol-β-cellobiose (18.6%), demonstrated the maximum xylanase and glucanase activities at pH 4.5–5.0 and 75 °C, retained stability over the pH range of 2.0–7.5 and at 70 °C, and was resistant to pepsin and trypsin, most metal ions and SDS. Multiple sequence alignment and modeled-structure analysis identified a unique Gly48 in Gt Xyn10, and site-directed mutagenesis of Gly48 to Lys improved the temperature optimum up to 80 °C. Under simulated mashing conditions, Gt Xyn10 (80 U) reduced the mash viscosity by 12.8% and improved the filtration rate by 31.3%. All these properties above make Gt Xyn10 attractive for potential applications in the feed and brewing industries. [ABSTRACT FROM AUTHOR]

[Display omitted] • Enzyme-polymer conjugates were first applied in W/W droplets for biocatalysis. • The effect of dimensionless numbers (We , Ca) on droplet size in jetting pattern were analyzed. • The power-law relationship between droplet size and velocity ratio of two-phase was clarified. • The K m value of microfluidic immobilized enzymatic was 1/2 of that in the batch reactor. • Xylanase embedded in the d-phase was reused 10 times with a residual activity above 60%. To gain a high yield of xylooligosaccharides (XOS) at a low cost, W/W droplet-based microfluidic system combined with immobilized xylanase was constructed. Jetting-regime under the high viscosity of both two phases was correlated with the Ca and We numbers. W/W droplets were obtained with controllable sizes when the flow velocities of dispersed and continuous phases were in the range of 0.29–0.48 µL/min and 0.52–0.80 µL/min respectively. The droplet size could be expressed as a power-law relationship with the velocity ratio of two phases. The maximum yield of XOS was 75.0% within 10 min. K m value was reduced to a half compared with batch reactors. Xylanase-polymer conjugates were reused for 10 times, with a residual activity over 60%. The results confirmed that the combination of enzyme-polymer conjugates and W/W droplet-based microfluidic system is a low-cost and high catalytic efficiency method for biocatalysis. [ABSTRACT FROM AUTHOR]

• Surplus mulberry residue was reused as a feedstock for sweet-flavored peptides production. • Multifrequency countercurrent ultrasonic technology was used to improve DMSP yield. • Established kinetic model can be used to predict treatment condition (R 2 = 0.9785, p > 0.05). • DMSPH was used to prepare flavored peptides that were eight-times sweeter than sucrose. • DMSPH had antioxidant activity and could retard growth of HepG2 cells in vitro. To increase the reuse of food residues, multifrequency countercurrent ultrasonic treatment was used to enhance the extraction yield of defatted mulberry seed protein (DMSP), and sweet-flavored peptides from DMSP hydrolysates (DMSPHs) were obtained for the first time. Here, the DMSP yield was increased by 16.2% (p < 0.05) while the power density was halved compared with single-frequency ultrasonic treatment. According to Fick's second law, a molecular diffusion dynamics model was developed to be suitable for predicting the pretreatment conditions (R 2 = 0.9785). After that, the sweet-flavored peptides were purified and the main amino acid sequences were identified, i.e., FEGGSIE, KDFPEAHSQAT, and GSQPAEGAK. Moreover, the antioxidant activities of DMSPHs prepared with tri-frequency treatment was higher than 60%. The DMSPHs retarded the growth of HepG2 cells in vitro, increased the necrotic quadrant (Q1-UL), and extended the S phase. Therefore, the sweet-flavored peptides prepared from DMSPHs using the multifrequency-ultrasonic treatment have significant biological activities. [ABSTRACT FROM AUTHOR]

[Display omitted] • Carrier-free immobilized xylanase was firstly constructed by ATRP "grafting-from" method. • Polymer grafting effectively increased the thermal stability (90 °C) of xylanase by 20 %. • The recycling of XYL-PNIPAAm conjugates was increased by 5 times (>60 %). • Over 99.95 % XOS in the DP range of 1–3 was produced from mulberry bark. In the past few years, xylooligosaccharides produced from lignocellulosic biomass have attracted great attention. Enzyme catalysis is widely used because of its mild reaction conditions. In order to solve the shortcomings of poor stability and high cost of enzyme, a carrier-free enzyme immobilization technology was introduced. For this purpose, protein-polymer conjugates, kinds of carrier-free immobilized enzymes, were first established by growing poly (N -isopropylacrylamide) chains on xylanase using a grafting-from method, to obtain reusable and high-value xylanase. SDS-PAGE and circular dichroism spectra showed the successful preparation of xylanase-PNIPAAm conjugates, and the integrity of the protein secondary structure. The remaining activity of the xylanase-PNIPAAm conjugates after heat treatment (90 °C) for 60 min was approximately 20 % higher than that of the free xylanase, and a higher pH stability was obtained in the pH range of 2−8. The remaining activity of xylanase-PNIPAAm conjugates was more than 60 % after recycling 5 times. In addition, the mulberry bark synergistic treated by xylanase-PNIPAAm conjugates and cellulase was studied. More than 99.95 % yield of XOS in the degree of polymerization range of 1–3 was produced from mulberry bark. Overall, the construction of xylanase-PNIPAAm conjugates provides a novel method to produce xylooligosaccharides from waste biomass. [ABSTRACT FROM AUTHOR]

• A novel microfluidic parallel-laminar flow ATPS with immobilized enzyme was constructed. • C 3 G content in mulberry red pigments was firstly improved by enzymatic hydrolysis of C 3 R. • C 3 G purity in the product was 88.78 % within 8.59 s, which was 68 % higher than the original. • Immobilized enzyme for MATPS could be reused 7 times with a relative activity above 60 %. • A new kind of mulberry red pigments with high purity of C 3 G close to 90 % was produced. Mulberry red pigments are safe, nontoxic and are one of the 48 natural pigments officially approved in China. Cyanidin-3- O -glucoside accounts for about 20 % of the total mulberry red pigments and has high physiological activity. Therefore, a novel microfluidic aqueous two-phase system with immobilized enzyme was constructed to enhance the cyanidin-3- O -glucoside content in red pigments under parallel-laminar flow condition for the first time. The effects of two-phase flow rates, temperature, pH and substrate concentration on cyanidin-3- O -rutinoside conversion and cyanidin-3- O -glucoside purity were investigated. The cyanidin-3- O -rutinoside conversion was 75.66 % within 8.59 s, and the purity of cyanidin-3- O -glucoside in the product was 88.78 % with an increase of 68 %. The immobilized enzyme could be reused seven times, maintaining a relative activity over 60 %. Compared with aqueous two-phase system, the overall reaction efficiency of mulberry red pigments in microfluidic aqueous two-phase system was higher. Cyanidin-3- O -rutinoside and cyanidin-3- O -glucoside were identified by HPLC-PDA-ESI-MS/MS. Overall, a new kind of high-quality red mulberry pigments with a cyanidin-3- O -glucoside purity close to 90 % was prepared using the innovative system quickly and effectively. This is of great significance for resource utilization. [ABSTRACT FROM AUTHOR]