Search

In this study, fourteen types of biochar produced using seven biomasses at temperatures 300 °C and 600 °C were screened for phenolics (furfural and hydroxymethylfurfural (HMF)) removal. Eucheuma spinosum biochar (EB-BC 600) showed higher adsorption capacity to furfural (258.94±3.2 mg/g) and HMF (222.81±2.3 mg/g). Adsorption kinetics and isotherm experiments interpreted that EB-BC 600 biochar followed the pseudo-first-order kinetic and Langmuir isotherm model for both furfural and HMF adsorption. Different hydrolysates were detoxified using EB-BC 600 biochar and used as feedstock for engineered Escherichia coli. An increased polyhydroxyalkanoates (PHA) production with detoxified barley biomass hydrolysate (DBBH: 1.71±0.07 g PHA/L), detoxified miscanthus biomass hydrolysate (DMBH: 0.87±0.03 g PHA/L) and detoxified pine biomass hydrolysate (DPBH: 1.28±0.03 g PHA/L) was recorded, which was 2.8, 6.4 and 3.4 folds high as compared to undetoxified hydrolysates. This study reports the mechanism involved in furfural and HMF removal using biochar and valorization of hydrolysate into PHA.

Poly(3-hydroxybutyrate) (PHB) is a biobased and biodegradable plastic. Considering the environmental issues of petroleum-based plastics, PHB is promising as it can be degraded in a relatively short time by bacteria to water and carbon dioxide. Substantial efforts have been made to identify PHB-degrading bacteria. To identify PHB-degrading bacteria, solid-based growth or clear zone assays using PHB as the sole carbon source are the easiest methods; however, PHB is difficult to dissolve and distribute evenly, and bacteria grow slowly on PHB plates. Here, we suggest an improved PHB plate assay using cell-grown PHB produced by Halomonas sp. and recovered by sodium dodecyl sulfate (SDS). Preparation using SDS resulted in evenly distributed PHB plates that could be used for sensitive depolymerase activity screening in less time compared with solvent-melted pellet or cell-grown PHB. With this method, we identified 15 new strains. One strain, Cutibacterium sp. SOL05 (98.4% 16S rRNA similarity to Cutibacterium acne), showed high PHB depolymerase activity in solid and liquid conditions. PHB degradation was confirmed by clear zone size, liquid culture, scanning electron microscopy, and Fourier-transform infrared spectroscopy. The results indicate this method can be used to easily identify PHB-degrading bacteria from various sources to strengthen the benefits of bioplastics.

Among various species of marine bacteria, those belonging to the genus Halomonas have several promising applications and have been studied well. However, not much information has been available on their antibiotic resistance. In our efforts to learn about the antibiotic resistance of strain Halomonas socia CKY01, which showed production of various hydrolases and growth promotion by osmolytes in previous study, we found that it exhibited resistance to multiple antibiotics including kanamycin, ampicillin, oxacillin, carbenicillin, gentamicin, apramycin, tetracycline, and spectinomycin. However, the H. socia CKY01 resistance pattern to kanamycin, gentamicin, apramycin, tetracycline, and spectinomycin differed in the presence of 10% NaCl and 1% NaCl in the culture medium. To determine the mechanism underlying this NaCl concentration-dependent antibiotic resistance, we compared four aminoglycoside resistance genes under different salt conditions while also performing time-dependent reverse transcription PCR. We found that the aph2 gene encoding aminoglycoside phosphotransferase showed increased expression under the 10% rather than 1% NaCl conditions. When these genes were overexpressed in an Escherichia coli strain, pETDuet-1::aph2 showed a smaller inhibition zone in the presence of kanamycin, gentamicin, and apramycin than the respective control, suggesting aph2 was involved in aminoglycoside resistance. Our results demonstrated a more direct link between NaCl and aminoglycoside resistance exhibited by the H. socia CKY01 strain.

Cadaverine, 1,5-diaminopentane, is one of the most promising chemicals for biobased-polyamide production and it has been successfully produced up to molar concentration. Pyridoxal 5′-phosphate (PLP) is a critical cofactor for inducible lysine decarboxylase (CadA) and is required up to micromolar concentration level. Previously the regeneration of PLP in cadaverine bioconversion has been studied and salvage pathway pyridoxal kinase (PdxY) was successfully introduced; however, this system also required a continuous supply of adenosine 5′-triphosphate (ATP) for PLP regeneration from pyridoxal (PL) which add in cost. Herein, to improve the process further a method of ATP regeneration was established by applying baker’s yeast with jhAY strain harboring CadA and PdxY, and demonstrated that providing a moderate amount of adenosine 5′-triphosphate (ATP) with the simple addition of baker’s yeast could increase cadaverine production dramatically. After optimization of reaction conditions, such as PL, adenosine 5′-diphosphate, MgCl2, and phosphate buffer, we able to achieve high production (1740 mM, 87% yield) from 2 M l-lysine. Moreover, this approach could give averaged 80.4% of cadaverine yield after three times reactions with baker’s yeast and jhAY strain. It is expected that baker’s yeast could be applied to other reactions requiring an ATP regeneration system.

Poly(3-hydroxybutyrate) (PHB) is a common polyhydroxyalkanoate (PHA) with potential as an alternative for petroleum-based plastics. Previously, we reported a new strain, Halomonas sp. YLGW01, which hyperproduces PHB with 94% yield using fructose. In this study, we examined the PHB production machinery of Halomonas sp. YLGW01 in more detail by deep-genome sequencing, which revealed a 3,453,067-bp genome with 65.1% guanine-cytosine content and 3054 genes. We found two acetyl-CoA acetyltransferases (Acetoacetyl-CoA thiolase, PhaA), one acetoacetyl-CoA reductase (PhaB), two PHB synthases (PhaC1, PhaC2), PHB depolymerase (PhaZ), and Enoyl-CoA hydratase (PhaJ) in the genome, along with two fructose kinases and fructose transporter systems, including the phosphotransferase system (PTS) and ATP-binding transport genes. We then examined the PHB production by Halomonas sp. YLGW01 using high-fructose corn syrup (HFCS) containing fructose, glucose, and sucrose in sea water medium, resulting in 7.95 ± 0.11 g/L PHB (content, 67.39 ± 0.34%). PHB was recovered from Halomonas sp. YLGW01 using different detergents; the use of Tween 20 and SDS yielded micro-sized granules with high purity. Overall, these results reveal the distribution of PHB synthetic genes and the sugar utilization system in Halomonas sp. YLGW01 and suggest a possible method for PHB recovery.

Psychrophilic bacteria, living at low and mild temperatures, can contribute significantly to our understanding of microbial responses to temperature, markedly occurring in the bacterial membrane. Here, a newly isolated strain, Pseudomonas sp. B14-6, was found to dynamically change its unsaturated fatty acid and cyclic fatty acid content depending on temperature which was revealed by phospholipid fatty acid (PLFA) analysis. Genome sequencing yielded the sequences of the genes Δ-9-fatty acid desaturase (desA) and cyclopropane-fatty acid-acyl-phospholipid synthase (cfa). Overexpression of desA in Escherichia coli led to an increase in the levels of unsaturated fatty acids, resulting in decreased membrane hydrophobicity and increased fluidity. Cfa proteins from different species were all found to promote bacterial growth, despite their sequence diversity. In conclusion, PLFA analysis and genome sequencing unraveled the temperature-related behavior of Pseudomonas sp. B14-6 and the functions of two membrane-related enzymes. Our results shed new light on temperature-dependent microbial behaviors and might allow to predict the consequences of global warming on microbial communities.

Bacteria can evade antibiotics by acquiring resistance genes, as well as switching to a non-growing dormant state without accompanying genetic modification. Bacteria in this quiescent state are called persisters, and this non-inheritable ability to withstand multiple antibiotics is referred to as antibiotic tolerance. Although all bacteria are considered to be able to form antibiotic-tolerant persisters, the antibiotic tolerance of extremophilic bacteria is poorly understood. Previously, we identified the psychrotolerant bacterium

γ-Aminobutyric acid (GABA), an important fine chemical in pharmacotherapy and food industries, is used as a novel material in the nylon industry and has attracted attention for its potential application in large scale production. Search for new genes and strains, development of efficient reaction systems, such as fermentation and bioconversion, and use of cheap starting material like monosodium glutamate (MSG) can make GABA production using less expensive bulk chemicals possible. Therefore, in this study, we constructed a recombinant Escherichia coli whole-cell system for GABA production that expressed glutamate decarboxylase (GAD) from Lactobacillus brevis and used MSG as the starting material. We also optimized the reaction conditions for MSG to GABA conversion, such as citrate buffer concentration, pyridoxal 5′te concentration, temperature, MSG concentration, and cell density (OD600). The optimized whole-cell system converted MSG to GABA via seven repetitive cycles resulting in an average conversion rate of 86% (71.7 mM/h) within 42 h.

Naturally degradable bioplastic polyhydroxyalkanoate (PHA) is a promising biopolymer and its physical properties could be changed by introducing of different monomers such as 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyhexanoate (3HHx). To produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) P(3HB-co-3HV)) including a high fraction of hydroxyvalerate, we introduced ctfAB into engineered Escherichia coli YJ200 possessing a pLW487 vector containing bktB, phaB, and phaC under control of the trc promoter. To enhance the HV fraction of P(3HB-co-3HV), the optimal concentrations of propionate, which acts as a precursor of 3HV and isopropyl β-D-1-thiogalactopyranoside, were determined and found to be 0.1 mM and 0.3%, respectively. Under the optimized conditions, E. coli, YJ201 produced P(3HB-co-3HV) containing a large amount of 3HV. Comparison with other CoA transferases showed that CtfAB produced relatively lower molecular weight copolymers. This demonstrates the necessity of identifying additional different CoA transferases, because CoA transferase can affect both the monomer fraction and molecular weight of polymers.

The present study deals with the utilization of lignocellulosic hydrolysate-based carbon source for exopolysaccharide (EPS) production using newly reported marine Echinicola sediminis BBL-M-12. This bacterium produced 7.56 g L

In this study, a newly isolated halotolerant strain Oceanisphaera arctica YHY1, capable of hydrolyzing seafood processing waste chitin biomass, is reported. Microbial fuel cells fed with 1% chitin and 40 g L−1 as the optimum salt concentration demonstrated stable electricity generation until 216 h (0.228 mA/cm2). N-acetyl-D-glucosamine (GlcNAc) was the main by-product in the chitin degradation, reaching a maximum concentration of 192.01 mg g−1 chitin at 120 h, whereas lactate, acetate, propionate, and butyrate were the major metabolites detected in the chitin degradation. O. arctica YHY1 utilized the produced GlcNAc, lactate, acetate, and propionate as the electron donors to generate the electric current. Cyclic voltammetry (CV) investigation revealed the participation of outer membrane-bound cytochromes, with extracellular redox mediators partly involved in the electron transfer mechanism. Furthermore, the changes in structural and functional groups in chitin after degradation were analyzed using FTIR and XRD. Therefore, the ability of O. arctica YHY1 to utilize waste chitin biomass under high salinities can be explored to treat seafood processing brine or high salt wastewater containing chitin with concurrent electricity generation.

One of the advantages of microbial synthesis of polyhydroxyalkanoates (PHAs) is the production of diverse polymers with different properties by the addition of different monomers, such as 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyhexanoate (3HHx). Considering the number of possible variables, terpolymers can have more variations than copolymers. In this study, we aimed to synthesize the terpolymer P(3HB-co-3HV-co-3HHx) from volatile fatty acids such as propionate and butyrate using the recombinant Ralstonia eutropha strain (Re2133/pCB81), containing deletions in the phaB1, phaB2, and phaB3 genes, and overexpression of synthetic PHA operon (phaC2, phaA, phaJ). This strain produced terpolymers depending on the ratio of two different carbon sources, namely, propionic and butyric acids; however, wild type R. eutropha could not produce the same type of polymer. The incorporation of 3-hydroxyvalerate and 3-hydroxyhexanoate monomers was confirmed by gas chromatography and H-nuclear magnetic resonance spectroscopy, and the parameters affecting the terpolymer composition were obtained based on regression. In addition, the thermal analysis showed that this terpolymer has properties different from those of the copolymer, obtained from the same composition of volatile acids. Depending on the ratio of two volatile acids, the composition of the terpolymer can be regulated resulting in different properties.

Streptomyces spp. have been isolated from different environmental niches and are known to exhibit diversity in secondary metabolism. They have complex regulation system to control secondary metabolism, however it is difficult to prioritize the importance of specific regulator among the thousands of global or cluster-situated genes that may regulate secondary metabolism in different Streptomyces strains. Here we suggest a simple homology-based selection method to find out important regulators by comparing seven Streptomyces strains finding highly homologous regulators in various Streptomyces strains and showed highly homologous 11 regulators containing four well known regulators such as BldM, IclR, WhiD and NdgR. Among various regulators, we showed a putative transcriptional regulator in Streptomyces coelicolor A3(2), SCO1463 playing a pivotal role in growth, antibiotic production (actinorhodin[ACT] and undecylprodigiosin [RED] production), and production/utilization of organic acids such as propionate and succinate by making comparisons between the deletion mutant and the wild type strain. Although high homology in various strains does not always mean the importance of a gene, we suggested a criterion on which regulator should be studied first and which would be more important among more than one thousand regulators.

Cadaverine, also known as 1,5-pentanediamine, is an important platform chemical with a wide range of applications and can be produced either by fermentation or bioconversion. Bioconversion of cadaverine from l -lysine is the preferred method because of its many benefits, including rapid reaction time and an easy downstream process. In our previous study, we replaced pyridoxal-5-phosphate (PLP) with pyridoxal kinase (PdxY) along with pyridoxal (PL) because it could achieve 80% conversion with 0.4 M of l -lysine in 6 h. However, conversion was sharply decreased in the presence of high concentrations of l -lysine (i.e., 1 M), resulting in less than 40% conversion after several hours. In this study, we introduced an ATP regeneration system using polyphosphate kinase (ppk) into systems containing cadaverine decarboxylase (CadA) and PdxY for a sufficient supply of PLP, which resulted in enhanced cadaverine production. In addition, to improve transport efficiency, the use of surfactants was tested. We found that membrane permeabilization via hexadecyltrimethylammonium bromide (CTAB) increased the yield of cadaverine in the presence of high concentrations of l -lysine. By combining these two strategies, the ppk system and addition of CTAB, we enhanced cadaverine production up to 100% with 1 M of l -lysine over the course of 6 h.

Food waste-derived volatile fatty acids (VFAs) can act as a renewable feedstock for biodiesel production. In synthetic media, Rhodococcus sp. YHY01 was able to utilize various organic acids (acetate, butyrate, lactate, and propionate) as a carbon source. Butyrate was the optimal carbon source, having a minimum inhibitory effect on growth, and a maximum growth yield coefficient (Yx/s 0.288 g dcw/g butyrate) and fatty acid yield coefficient (Yf/s 0.206 g/g butyrate), compared to other organic acids (lactate, propionate, and acetate). Acetate, butyrate, and lactate mostly supported the production of fatty acids with an even number of carbons, whereas propionate enhanced the content of odd-numbered fatty acids. Response surface methodology (RSM) design study resulted in maximum biomass (2.8 g/L) and fatty acid yield (1.9 g/g) with acetate:butyrate:lactate (0.333:0.333:0.333) as a carbon source. Culture of Rhodococcus sp. YHY01 in media containing food waste-derived VFAs as the carbon source had a biomass (3.2 g dcw/L), fatty acid yield (2.2 g/L), and fatty acid accumulation (69% w/w) under nitrogen-limited condition. Biodiesel produced from food waste had an iodine value (IV, 37), cetane number (CN, 63), high heating value (HHV, 39), density (υ, 3.9), and viscosity (ρ, 0.868) that meet international standards.

Polyhydroxybutyrate (PHB), the most well-known polyhydroxyalkanoate, is a bio-based, biodegradable polymer that has the potential to replace petroleum-based plastics. Lignocellulose hydrolysate, a non-edible resource, is a promising substrate for the sustainable, fermentative production of PHB. However, its application is limited by the generation of inhibitors during the pretreatment processes. In this study, we investigated the feasibility of PHB production in E. coli in the presence of inhibitors found in lignocellulose hydrolysates. Our results show that the introduction of PHB synthetic genes (bktB, phaB, and phaC from Ralstonia eutropha H16) improved cell growth in the presence of the inhibitors such as furfural, 4-hydroxybenzaldehyde, and vanillin, suggesting that PHB synthetic genes confer resistance to these inhibitors. In addition, increased PHB production was observed in the presence of furfural as opposed to the absence of furfural, suggesting that this compound could be used to stimulate PHB production. Our findings indicate that PHB production using lignocellulose hydrolysates in recombinant E. coli could be an innovative strategy for cost-effective PHB production, and PHB could be a good target product from lignocellulose hydrolysates, especially glucose.

A novel marine bacterium Arenibacter palladensis YHY2 isolated from Eastern Sea, South Korea revealed hydrolysing ability of crab chitin in shake flask and microbial fuel cell (MFC) systems. Under shake flask cultivation, strain YHY2 demonstrated 71.44 ± 1.90 U/ml chitinase activity with the microbial cell optical density (OD) 0.699 ± 0.021. High performance liquid chromatography (HPLC) results showed N-acetylglucosamine (GlcNAc) as the major by-product of chitin degradation. Furthermore, Ralstonia eutropha H16 which has carbon utilization limited to GlcNAc and fructose was co-cultivated with YHY2 to consume GlcNAc and accumulate polyhydroxyalkanoates (PHA). However, on co-cultivating strain YHY2 with H16 surprisingly showed higher production of chitinase (97.89 ± 2.72 U/ml) and GlcNAc (80.31 mg GlcNAc/g chitin) corresponding to the microbial cell OD (1.065 ± 0.005) at 120 h. In addition, marine strain YHY2 demonstrated biofilm formation, hence co-cultivation of strain YHY2 and H16 under MFC system was performed to check the electricity production. Maximum electricity current output density was 15.15 μA/cm2 at the initial stage and 10.72 μA/cm2 at 204 h. GlcNAc (196.34 mg GlcNAc/g chitin) was produced along with other metabolites like butyrate, acetate, lactate, and propionate in MFC system. The microbial cell biomass from MFC was analysed for PHA content showed 1.020 g/l of 3-polyhydroxybutyrate (PHB) and 0.198 g/l of 3-polyhydroxyvalerate (PHV) as the main components of PHA. Moreover, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM) were performed to reveal changes in chitin before and after degradation. Thus, the exploitation of recalcitrant chitin biomass for simultaneous production of GlcNAc, electricity, and PHA in one-pot MFC system was the productive asset for sustainable development.

Polyhydroxybutyrates (PHB) are biodegradable polymers that are produced by various microbes, including Ralstonia, Pseudomonas, and Bacillus species. In this study, a Vibrio proteolyticus strain, which produces a high level of polyhydroxyalkanoate (PHA), was isolated from the Korean marine environment. To determine optimal growth and production conditions, environments with different salinity, carbon sources, and nitrogen sources were evaluated. We found that the use of a medium containing 2% (w/v) fructose, 0.3% (w/v) yeast extract, and 5% (w/v) sodium chloride (NaCl) in M9 minimal medium resulted in high PHA content (54.7%) and biomass (4.94 g/L) over 48 h. Addition of propionate resulted in the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(HB-co-HV)) copolymer as propionate acts as a precursor for the HV unit. In these conditions, the bacteria produced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) containing a 15.8% 3HV fraction with 0.3% propionate added as the substrate. To examine the possibility of using unsterilized media with high NaCl content for PHB production, V. proteolyticus was cultured in sterilized and unsterilized conditions. Our results indicated a higher growth, leading to a dominant population in unsterilized conditions and higher PHB production. This study showed the conditions for halophilic PHA producers to be later implemented at a larger scale.

The existing study deals with adsorptive removal of the endocrine-disrupting chemical bisphenol-A and toxic azo dye solvent black-3 from single and binary solutions. These two chemicals are commonly used as an additive in the synthetic plastic industries. Among the tested twenty pristine and modified biochars, the pristine pinecone biochar produced at 750 °C revealed greater bisphenol-A removal. Simulation of the experimental data obtained for bisphenol-A and dye removal from the single-component solution offered a best-fit to Elovich (R

Polyhydroxyalkanoates (PHAs) are attractive new bioplastics for the replacement of plastics derived from fossil fuels. With their biodegradable properties, they have also recently been applied to the medical field. As poly(3-hydroxybutyrate) produced by wild-type Ralstonia eutropha has limitations with regard to its physical properties, it is advantageous to synthesize co- or terpolymers with medium-chain-length monomers. In this study, tung oil, which has antioxidant activity due to its 80% α-eleostearic acid content, was used as a carbon source and terpolymer P(53 mol% 3-hydroxybytyrate-co-2 mol% 3-hydroxyvalerate-co-45 mol% 3-hydroxyhexanoate) with a high proportion of 3-hydroxyhexanoate was produced in R. eutropha Re2133/pCB81. To avail the benefits of α-eleostearic acid in the tung oil-based medium, we performed partial harvesting of PHA by using a mild water wash to recover PHA and residual tung oil on the PHA film. This resulted in a film coated with residual tung oil, showing antioxidant activity. Here, we report the first application of tung oil as a substrate for PHA production, introducing a high proportion of hydroxyhexanoate monomer into the terpolymer. Additionally, the residual tung oil was used as an antioxidant coating, resulting in the production of bioactive PHA, expanding the applicability to the medical field.

The present investigation deals with the adsorptive removal of crude petroleum oil from the water surface using coconut oil-modified pinewood biochar. Biochar generated at higher pyrolysis temperature (700 °C) revealed higher fatty acid-binding efficiency responsible for the excellent hydrophobicity of the biochar. Fatty acids composition attached to the biochar produced at 700 °C was (mg g−1 BC) lauric acid (9.024), myristic acid (5.065), palmitic acid (2.769), capric acid (1.639), oleic acid (1.362), stearic acid (1.114), and linoleic acid (0.130). Simulation of the experimental adsorption data of pristine and modified pinewood biochar generated at 700 °C offered the best fit to pseudo-first-order kinetics (R2 > 0.97) and Langmuir isotherm model (R2 > 0.99) based on the highest regression coefficients. Consequently, the adsorption process was mainly driven by surface hydrophobic interactions including π-π electron-donor-acceptor between electron-rich (π-donor) polycyclic aromatic hydrocarbons from the crude oil and biochar (π-acceptor). A maximum adsorption capacity (Qmax) of 5.315 g g−1 was achieved by modified floating biochar within 60 min. Whereas the reusability testing revealed 49.39% and 51.40% was the adsorption efficiency of pristine and modified biochar at the fifth adsorption-desorption cycle.

In the present study, an exopolysaccharide (EPS)-producing bacterial strain was isolated from the Eastern Sea (Sokcho Beach) of South Korea and identified as Sphingobium yanoikuyae BBL01. Media optimization was performed using response surface design, and a yield of 2.63 ± 0.02 g/L EPS was achieved. Purified EPS produced using lactose as the main carbon source was analyzed by GC–MS and found to be composed of α-D-xylopyranose (28.6 ± 2.0%), β-D-glucopyranose (21.0 ± 1.6%), α-D-mannopyranose (18.5 ± 1.2%), β- d -mannopyranose (13.1 ± 1.4%), β-D-xylopyranose (10.2 ± 2.1%), α- d -talopyranose (5.9 ± 1.1%), and β- d -galacturonic acid (2.43 ± 0.8%). Interestingly, different carbon sources (glucose, galactose, glycerol, lactose, sucrose, and xylose) showed no effect on EPS monomer composition, with a slight change in the mass percentage of various monosaccharides. Purified EPS was stable up to 233 °C, indicating its possible suitability as a thickening and gelling agent for food-related applications. EPS also showed considerable emulsifying, flocculating, free-radical scavenging, and metal-complexion activity, suggesting various biotechnological applications.

Among various species of marine bacteria, those belonging to the genus

The present study aimed towards adsorptive removal of the toxic azo dye onto biochar derived from Eucheuma spinosum biomass. Characterization of the produced biochar was performed using X-ray diffraction (XRD), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), and Brunauer-Emmett-Teller (BET). Eucheuma spinosum biochar (ES-BC) produced at 600 °C revealed a maximum adsorption capacity of 331.97 mg/g towards reactive red 120 dye. The adsorption data fitted best to the pseudo-second order kinetics (R2 > 0.99) and Langmuir isotherm (R2 > 0.98) models. These adsorption models signified the chemisorption mechanism with monolayer coverage of the adsorbent surface with dye molecules. Furthermore, the adsorption process was mainly governed by electrostatic interaction, ion exchange, metal complexation, and hydrogen bonding as supported by the solution pH, FTIR, XPS, and XRD investigation. Nevertheless, alone adsorption technology could not offer a complete solution for eliminating the noxious dyes. Therefore, the bioelectrochemical system (BES) equipped with previously isolated marine Shewanella marisflavi BBL25 was intended for the complete remediation of azo dye. The BES II demonstrated highest dye decolorization (97.06%) within 48 h at biocathode where the reductive cleavage of the azo bond occurred. Cyclic voltammetry (CV) studies of the BES revealed perfect redox reactions taking place where the redox mediators shuttled the electrons to the dye molecule to accelerate the dye decolorization. Besides, the GC-MS analysis revealed biotransformation of the dye into less toxic metabolites as tested using a phyto and cytogenotoxicity.

Phenol-soluble modulins (PSMs) are responsible for regulating biofilm formation, persister cell formation

Pipecolic acid, a non-proteinogenic amino acid, is a metabolite in lysine metabolism and a key chiral precursor in local anesthesia and macrolide antibiotics. To replace the environmentally unfriendly chemical production or preparation procedure of pipecolic acid, many biological synthetic routes have been studied for a long time. Among them, synthesis by lysine cyclodeaminase (LCD), encoded by pipA, has several advantages, including stability of enzyme activity and NAD+ self-regeneration. Thus, we selected this enzyme for pipecolic acid biosynthesis in a whole-cell bioconversion. To construct a robust pipecolic acid production system, we investigated important conditions including expression vector, strain, culture conditions, and other reaction parameters. The most important factor was the introduction of multiple pipA genes into the whole-cell system. As a result, we produced 724 mM pipecolic acid (72.4 % conversion), and the productivity was 0.78 g/L/h from 1 M l-lysine after 5 days. This is the highest production reported to date.

Bacteria from the genus Halomonas hold promise in biotechnology as sources of salt-tolerant enzymes, biosurfactants, biopolymers, osmolytes, and as actors in bioremediation processes. In a previous work, we have identified Halomonas socia strain CKY01 having various hydrolase activities. Here, we aimed to study the survival strategies of marine bacteria. A deep genome sequencing study of H. socia CKY01 has revealed 4627 genes reaching 4,753,299 bp with 64 % of GC content. This strain produced polyhydroxybutyrate (PHB) having one gene clusters having phaC and phasin, and it has several genes responsible for the uptake, synthesis, and transport of the osmolytes such as betaine, choline, ectoine, carnitine, and proline in the bacterial genome. The addition of 60 mM glutamate, 60 mM proline and 60 mM ectoine enhanced growth 300.8 %, 294.2 % and 235.0 %, respectively, under 10 % saline conditions. In particular, ectoine and proline increased salt resistance and allowed cells to survive in more than 15 % NaCl. By combining experimental and genome sequencing data, we have investigated the importance of osmolytes on the survival of this Halomonas strain.

MRSA is Methicillin-resistantStaphylococcus aureusand they are widespread and making trouble in treatment in communities and surgical areas. MRSA have been adapted to antibiotics so that they can block the access of antibiotics physically or chemically deactivate it or modify the precursor of the target. Flavonoids are secondary metabolites which are naturally produced by plant or fungus and they are acting generally as pigment, quorum sensing molecules, antibiotics to other competitive microorganisms. Their natural origins and multiple activities have drawn much attention to be developed as the potential drugs since flavonoids could be a good candidate to overcome antibiotic resistant bacteria. Among various flavonoids, we found out naringenin has antibacterial activity on MRSA and Δagrmutants which are more resistance than MRSA to beta-lactam antibiotics by decreasing biofilm formation dramatically and decreasing the secretion of fatty acid. It also showed high synergetic activity with oxacillin to both antibacterial activity and biofilm inhibition. Considering the number of flavonoids, our experiments expand the possibility of the use of flavonoids to MRSA.

Additional file 1. Additional tables and figures.

Glutaric acid is one of the promising C5 platform compounds in the biochemical industry. It can be produced chemically, through the ring-opening of butyrolactone followed by hydrolysis. Alternatively, glutaric acid can be produced via lysine degradation pathways by microorganisms. In microorganisms, the overexpression of enzymes involved in this pathway from E. coli and C. glutamicum has resulted in high accumulation of 5-aminovaleric acid. However, the conversion from 5-aminovaleric acid to glutaric acid has resulted in a relatively low conversion yield for unknown reasons. In this study, as a solution to improve the production of glutaric acid, we introduced gabTD genes from B. subtilis to E. coli for a whole cell biocatalytic approach. This approach enabled us to determine the effect of co-factors on reaction and to achieve a high conversion yield from 5-aminovaleric acid at the optimized reaction condition. Optimization of whole cell reaction by different plasmids, pH, temperature, substrate concentration, and cofactor concentration achieved full conversion with 100 mM of 5-aminovaleric acid to glutaric acid. Nicotinamide adenine dinucleotide phosphate (NAD(P)+) and α-ketoglutaric acid were found to be critical factors in the enhancement of conversion in selected conditions. Whole cell reaction with a higher concentration of substrates gave 141 mM of glutaric acid from 300 mM 5-aminovaleric acid, 150 mM α-ketoglutaric acid, and 60 mM NAD+ at 30 °C, with a pH of 8.5 within 24 h (47.1% and 94.2% of conversion based on 5-aminovaleric acid and α-ketoglutaric acid, respectively). The whole cell biocatalyst was recycled 5 times with the addition of substrates; this enabled the accumulation of extra glutaric acid.

Polyhydroxyalkanoates (PHA), such as poly (3-hydroxybutyrate) (PHB), have emerged as potential alternatives to petroleum-based plastics and can be produced through the appropriate selection of marine bacteria that are already adapted to high salt and low temperature conditions without the requirement of antibiotic treatment. The present study, thus, aimed to screen and characterize thirteen PHA-producing microbial strains isolated from the Gwangalli beach in Busan, South Korea. Among them, Halomonas sp. YLGW01 produced the highest amount of PHB (94.6 ± 1.8% (w/w)) using fructose. Interestingly Halomonas sp. YLGW01 showed increase in cell size (8.39 ± 3.63 μm) with fructose as carbon source as compared to glucose (2.34 ± 0.44 μm). Fructose syrup was investigated as carbon source under unsterilized conditions and 95.26 ± 1.78% of PHB was produced. Overall, this strain showed the highest PHB contents in halotolerant bacteria.

In this study, a heterogeneous catalyst prepared by pyrolysis of waste cork (Quercus suber) was used for the transesterification of waste cooking oil (WCO). Physicochemical properties of the synthesized biochar catalyst were studied using BET, SEM, FTIR, and XRD. The experiment results demonstrate that heterogeneous catalyst synthesized at 600 °C showed maximum fatty acids methyl esters (FAMEs) conversion (98%) at alcohol:oil (25:1), catalyst loading (1.5% w/v) and temperature 65 °C. Biodiesel produced from WCO (Canola oil) mainly composed of FAMEs in following order C18:1 > C18:2 > C16:0 > C18:0 > C20:0. Properties of produced biodiesel were analysed as cetane number (CN) 50.56, higher heating value (HHV) 39.5, kinematic viscosity (ʋ) 3.9, and density (ρ) 0.87.

Itaconic acid is an important organic acid and a major component of various polymers. It is used in resins, superabsorbent polymers, and substitutes for petrochemicalbased monomers such as acrylic and methacrylic acids. Itaconic acid is primarily produced by the fungus Aspergillus terreus, which yields a high titer with albeit long fermentation period and by-products. In our previous study, Escherichia coli JY001 was reported to produce itaconic acid using citric acid in whole-cell reaction resulting in higher itaconic acid productivity with less by-products formation. The present study aimed to increase whole-cell enzyme stability and reusability, via immobilization of E. coli JY001 using barium-alginate beads. We optimized the cations, temperature, pH, alginate, BaCl2 concentration, cell density per bead, and CTAB content to improve transfer rate of substrates and products. Under the optimized conditions, immobilized whole cells were stable for four repeated cycles of itaconic acid production. The present results would strengthen the basis for a continuous itaconic acid production.

The cellular components of Akkermansia muciniphila are considered potential biotherapeutics for the improvement of obesity, diabetes, and metabolic diseases. However, the molecular-based mechanism of A. muciniphila for treatment of obesity, which can provide important evidence for human research, has rarely been explored. Here, we applied integrative multiomics approaches to investigate the underlying molecular mechanism involved in obesity treatment by A. muciniphila. First, the treatment with a cell lysate of A. muciniphila reduced lipid accumulation in 3T3-L1 cells and downregulated the mRNA expression of proteins involved in adipogenesis and lipogenesis. Our proteomic results revealed that A. muciniphila decreased the expression of proteins involved in fat cell differentiation, fatty acid metabolism, and energy metabolism in adipocytes. Moreover, A. muciniphila significantly reduced the level of metabolites related to glycolysis, the TCA cycle, and ATP in adipocytes. Interestingly, serine protease inhibitor A3 (SERPINA3) homologs were overexpressed in the 3T3-L1 cells treated with A. muciniphila. Small interfering RNA (siRNA) transfection demonstrated that A. muciniphila upregulates SERPINA3G expression and inhibits lipogenesis in adipocytes. Taken together, our multiomics-based approaches enabled to uncover the molecular mechanism of A. muciniphila for treatment of obesity and provide potent anti-lipogenic agents.

Petroleum-based plastic is a widely used commodity in modern living due to its extensive industrial and domestic applications. Polyethylene and polypropylene account for more than 50% of total plastic usage, and they resist decomposition in a natural environment. In this study, Lysinibacillus sp., isolated and identified as a novel strain, was investigated to decompose polyethylene and polypropylene. In the microbial cultivation medium without any physicochemical pretreatment, the Lysinibacillus sp. reduced the weight of polypropylene and polyethylene by approximately 4 and 9%, respectively, over 26 days. During the biodegradation of polyethylene and polypropylene, a rough surface increase was observed by scanning electron microscope (SEM) imaging. Moreover, various oxidation products containing CH2 groups produced from the decomposition of polyethylene and polypropylene were observed by GC–MS analysis. Results suggest that microorganisms can be used to degrade polyethylene and polypropylene without a physicochemical process. This can be a starting point for developing methods for remediating soil contaminated with plastics.

Once cells have been used to produce biochemicals, there are only a few effective ways to utilize the residual cell mass, even though the utilization of leftover cells would aid in decreasing production costs. Here, a polyhydroxybutyrate (PHB) and isobutanol co-production system was designed to address this challenge. The addition of the PHB operon into Escherichia coli conferred a metabolic advantage for alcohol production, generating 1.14-fold more isobutanol. Furthermore, following nitrogen source optimization and cofactor engineering, the engineered E. coli strain produced 2-fold more isobutanol and 0.25 g/L PHB. Moreover, E. coli cells showed higher tolerance to isobutanol with the overexpression of PHB biosynthesis genes. This co-production system resulted in an increased biomass, higher glucose utilization, and lower acetate maintenance, leading to higher productivity regarding PHB and isobutanol yield. Thus, this novel system is applicable to future fermentation studies for the co-production of PHB and isobutanol.

Bacteria have evolved a defense system to resist external stressors, such as heat, pH, and salt, so as to facilitate survival in changing or harsh environments. However, the specific mechanisms by which bacteria respond to such environmental changes are not completely elucidated. Here, we used halotolerant bacteria as a model to understand the mechanism conferring high tolerance to NaCl. We screened for genes related to halotolerance in Halomonas socia, which can provide guidance for practical application. Phospholipid fatty acid analysis showed that H. socia cultured under high osmotic pressure produced a high portion of cyclopropane fatty acid derivatives, encoded by the cyclopropane-fatty acid-acyl phospholipid synthase gene (cfa). Therefore, H. socia cfa was cloned and introduced into Escherichia coli for expression. The cfa-overexpressing E. coli strain showed better growth, compared with the control strain under normal cultivation condition as well as under osmotic pressure (> 3% salinity). Moreover, the cfa-overexpressing E. coli strain showed 1.58-, 1.78-, 3.3-, and 2.19-fold higher growth than the control strain in the presence of the inhibitors furfural, 4-hydroxybenzaldehyde, vanillin, and acetate from lignocellulosic biomass pretreatment, respectively. From a practical application perspective, cfa was co-expressed in E. coli with the polyhydroxyalkanoate (PHA) synthetic operon of Ralstonia eutropha using synthetic and biosugar media, resulting in a 1.5-fold higher in PHA production than that of the control strain. Overall, this study demonstrates the potential of the cfa gene to boost cell growth and production even in heterologous strains under stress conditions.

The present study enlightens facile synthesis and characterization of magnetic biochar derived from waste banana pseudostem biomass for the removal of nitrofuran antibiotic ‘furazolidone’ (FZD). Brunauer-Emmett-Teller (BET), scanning electron microscopy (SEM), magnetic hysteresis, X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) revealed successful hybridization of magnetic nanocomposites with biochar (BPB600). The maximum adsorption capacity of magnetic BPB600 was 96.81% (37.86 mg g−1), which was significantly higher than the non-coated BPB600 (77.25%; 31.45 mg g−1). Adsorption kinetics data fitted well with pseudo-second order, and Elovich model demonstrating dominance of the chemisorption mechanism. Furthermore, the response surface methodology (RSM) was applied to evaluate the interactive effect of pH, temperature, and FZD concentration on adsorption. Therefore, the results of present study would provide an effective strategy to tackle antibiotic contaminants responsible for the antibiotic resistance genes or bacteria that decreases the therapeutic value of antibiotics.

Rhodococcus sp. YHY01 was studied to utilize various lignin derived aromatic compounds. It was able to utilize p-coumaric acid, cresol, and 2,6 dimethoxyphenol and resulted in biomass production i.e. 0.38 g dcw/L, 0.25 g dcw/L and 0.1 g dcw/L, and lipid accumulation i.e. 49%, 40%, 30%, respectively. The half maximal inhibitory concentration (IC50) value for p-coumaric acid (13.4 mM), cresol (7.9 mM), and 2,6 dimethoxyphenol (3.4 mM) was analyzed. Dimethyl sulfoxide (DMSO) solubilized barley straw lignin fraction was used as a carbon source for Rhodococcus sp. YHY01 and resulted in 0.130 g dcw/L with 39% w/w lipid accumulation. Major fatty acids were palmitic acid (C16:0) 51.87%, palmitoleic acid (C16:l) 14.90%, and oleic acid (C18:1) 13.76%, respectively. Properties of biodiesel produced from barley straw lignin were as iodine value (IV) 27.25, cetane number (CN) 65.57, cold filter plugging point (CFPP) 14.36, viscosity (υ) 3.81, and density (ρ) 0.86.

Polyhydroxybutyrate (PHB), the most well-known polyhydroxyalkanoate, is a bio-based, biodegradable polymer that has the potential to replace petroleum-based plastics. Lignocellulose hydrolysate, a non-edible resource, is a promising substrate for the sustainable, fermentative production of PHB. However, its application is limited by the generation of inhibitors during the pretreatment processes. In this study, we investigated the feasibility of PHB production in

Polyhydroxyalkanoate (PHA) is a potential substitute for petroleum-based plastics and can be produced by many microorganisms, including recombinant Escherichia coli. For efficient conversion of substrates and maximum PHA production, we performed multiple engineering of branched pathways in E. coli. We deleted four genes (pflb, ldhA, adhE, and fnr), which contributed to the formation of byproducts, using the CRISPR/Cas9 system and overexpressed pntAB, which catalyzes the interconversion of NADH and NADPH. The constructed strain, HR002, showed accumulation of acetyl-CoA and decreased levels of byproducts, resulting in dramatic increases in cell growth and PHA content. Thus, we demonstrated the effects of multiple engineering for redirecting carbon flux into PHA production without any concerns regarding simultaneous deletion.

Odd chain fatty acids serve as anti-allergic, anti-inflammatory, and antifungal agents, and are useful for the production of biodiesel. Rhodococcus sp. YHY01 utilizes a wide range of carbon sources and accumulate lipids i.e. fructose (37% w/w dcw) glucose (56% w/w dcw), glycerol (50% w/w dcw), acetate (42% w/w dcw), butyrate (65% w/w dcw), lactate (56% w/w dcw), and propionate (62% w/w dcw). In this study, propionate was proved as the best carbon source and produced 69% odd chain fatty acids of total fatty acids, followed by glycerol (13% odd chain fatty acids of total fatty acids). A synthetic medium optimized with response surface design containing glycerol, propionate, and ammonium chloride (0.32%:0.76%:0.040% w/v) facilitated the production of total fatty acids 69% w/w of dcw, and odd chain fatty acids comprised 85% w/w of total fatty acids. Major odd chain fatty acids were in the order C17:0 > C15:0 > Cis-10-C17:1 > 10Me-C17:0 > C19:0 > Cis-10-C19:1.