Improvement of cadaverine production in whole cell system with baker's yeast for cofactor regeneration.

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.

Simultaneous monitoring of the bioconversion from lysine to glutaric acid by ethyl chloroformate derivatization and gas chromatography-mass spectrometry.

Glutaric acid is a precursor of a plasticizer that can be used for the production of polyester amides, ester plasticizer, corrosion inhibitor, and others. Glutaric acid can be produced either via bioconversion or chemical synthesis, and some metabolites and intermediates are produced during the reaction. To ensure reaction efficiency, the substrates, intermediates, and products, especially in the bioconversion system, should be closely monitored. Until now, high performance liquid chromatography (HPLC) has generally been used to analyze the glutaric acid-related metabolites, although it demands separate time-consuming derivatization and non-derivatization analyses. To substitute for this unreasonable analytical method, we applied herein a gas chromatography - mass spectrometry (GC-MS) method with ethyl chloroformate (ECF) derivatization to simultaneously monitor the major metabolites. We determined the suitability of GC-MS analysis using defined concentrations of six metabolites (l-lysine, cadaverine, 5-aminovaleric acid, 2-oxoglutaric acid, glutamate, and glutaric acid) and their mass chromatograms, regression equations, regression coefficient values (R2), dynamic ranges (mM), and retention times (RT). This method successfully monitored the production process in complex fermentation broth.

Effects of a Δ-9-fatty acid desaturase and a cyclopropane-fatty acid synthase from the novel psychrophile Pseudomonas sp. B14-6 on bacterial membrane properties.

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.

Enhanced tolerance to inhibitors of Escherichia coli by heterologous expression of cyclopropane-fatty acid-acyl-phospholipid synthase (cfa) from Halomonas socia.

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.

Enhanced production of glutaric acid by NADH oxidase and GabD-reinforced bioconversion from l-lysine.

Glutaric acid is a promising alternative chemical to phthalate plasticizer since it can be produced by the bioconversion of lysine. Though, recent studies have enabled the high-yield production of its precursor, 5-aminovaleric acid (AMV), glutaric acid production via the AMV pathway has been limited by the need for cofactors. Introduction of NAD(P)H oxidase (Nox) with GabTD enzyme remarkably diminished the demand for oxidized nicotinamide adenine dinucleotide (NAD+ ). Supply of oxygen through vigorous shaking had a significant effect on the conversion of AMV with a reduced requirement of NAD + . A high conversion rate was achieved in Nox coupled GabTD reaction under optimized expression vector, terrific broth (TB), and pH 8.5 at high cell density. Supplementary expression of GabD resulted in the production of 353 ± 35 mM glutaric acid with 88.3 ± 8.7% conversion from 400 mM AMV. Moreover, the reaction with a higher concentration of AMV could produce 528 ± 21 mM glutaric acid with 66.0 ± 2.7% conversion. In addition, the co-biotransformation strategy of GabTD and DavBA whole cells could produce 282 mM glutaric acid with 70.8% conversion from lysine, compared to the 111 mM glutaric acid yield from the combined GabTD-DavBA system.

Polyhydroxybutyrate production in halophilic marine bacteria Vibrio proteolyticus isolated from the Korean peninsula.

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.

An efficient and eco-friendly approach for the sustainable recovery and properties characterization of polyhydroxyalkanoates produced by methanotrophs.

Synthetic biodegradable and bio-based polymers have emerged as sustainable alternatives to nonrenewable petroleum-derived polymers which cause serious environmental issues. In particular, polyhydroxyalkanoates (PHA) are promising biopolymers owing to their outstanding biodegradability and biocompatibility. The production of the homopolymer poly(3-hydroxybutyrate) (PHB) and copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) from type II methanotrophs via microbial fermentation was presented. For the efficient extraction and recovery of intracellular PHA from methanotrophs, different extraction approaches were investigated including solvent extraction using 1,3-dioxolane as a green solvent, integrated cell lysis and solvent extraction, and cell digestion without the use of organic solvents. Among various extraction approaches, the integrated method exhibited the highest extraction performance, with PHA recovery and purity exceeding 91 % and 93 %, respectively, even when the PHA content of the cells was low. Furthermore, the molecular weight, thermal stability, and mechanical properties of the recovered PHA were comprehensively analyzed to suggest its suitable practical applications. The obtained properties were comparable to that of the commercial PHA products and PHA produced from other microbial species, indicating an efficient recovery of high-quality PHA produced from methanotrophs.

Tryptophan-Based Hyperproduction of Bioindigo by Combinatorial Overexpression of Two Different Tryptophan Transporters.

Indigo is a valuable, natural blue dye that has been used for centuries in the textile industry. The large-scale commercial production of indigo relies on its extraction from plants and chemical synthesis. Studies are being conducted to develop methods for environment-friendly and sustainable production of indigo using genetically engineered microbes. Here, to enhance the yield of bioindigo from an E. coli whole-cell system containing tryptophanase (TnaA) and flavin-containing monooxygenase (FMO), we evaluated tryptophan transporters to improve the transport of aromatic compounds, such as indole and tryptophan, which are not easily soluble and passable through cell walls. Among the three transporters, Mtr, AroP, and TnaB, AroP enhanced indigo production the most. The combination of each transporter with AroP was also evaluated, and the combination of AroP and TnaB showed the best performance compared to the single transporters and two transporters. Bioindigo production was then optimized by examining the culture medium, temperature, isopropyl ß-D-1-thiogalactopyranoside concentration, shaking speed (rpm), and pH. The novel strain containing aroP and tnaB plasmid with tnaA and FMO produced 8.77 mM (2.3 g/l) of bioindigo after 66 h of culture. The produced bioindigo was further recovered using a simple method and used as a watercolor dye, showing good mixing with other colors and color retention for a relatively long time. This study presents an effective strategy for enhancing indigo production using a combination of transporters.

Maximization of 3-hydroxyhexanoate fraction in poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) using lauric acid with engineered Cupriavidus necator H16.

As polyhydroxybutyrate (P(3HB)) was struggling with mechanical properties, efforts have been directed towards increasing mole fraction of 3-hydroxyhexanoate (3HHx) in P(3HB-co-3HHx) to improve the properties of polyhydroxyalkanoates (PHAs). Although genetic modification had significant results, there were several issues related to cell growth and PHA production by deletion of PHA synthetic genes. To find out easier strategy for high 3HHx mole fraction without gene deletion, Cupriavidus necator H16 containing phaC2Ra-phaACn-phaJ1Pa was examined with various oils resulting that coconut oil gave the highest 3HHx mole fraction. When fatty acid composition analysis with GC-MS was applied, coconut oil was found to have very different composition from other vegetable oil containing very high lauric acid (C12) content. To find out specific fatty acid affecting 3HHx fraction, different fatty acids from caproic acid (C6) to stearic acid (C18) was evaluated and the 3HHx mole fraction was increased to 26.5 ± 1.6 % using lauric acid. Moreover, the 3HHx mole fraction could be controlled from 9 % to 31.1 % by mixing bean oil and lauric acid with different ratios. Produced P(3HB-co-3HHx) exhibited higher molecular than P(3HB-co-3HHx) from phaB-deletion mutant. This study proposes another strategy to increase 3HHx mole fraction with easier way by modifying substrate composition without applying deletion tools.

Identification of Oil-Loving Cupriavidus necator BM3-1 for Polyhydroxyalkanoate Production and Assessing Contribution of Exopolysaccharide for Vegetable Oil Utilization.

Polyhydroxyalkanoates (PHA) have received attention owing to their biodegradability and biocompatibility, with studies exploring PHA-producing bacterial strains. As vegetable oil provides carbon and monomer precursors for poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(3HB-co-3HHx)), oil-utilizing strains may facilitate PHA production. Herein, Cupriavidus necator BM3-1, which produces 11.1 g/L of PHB with 5% vegetable oil, was selected among various novel Cupriavidus necator strains. This strain exhibited higher preference for vegetable oils over sugars, with soybean oil and tryptone determined to be optimal sources for PHA production. BM3-1 produced 33.9 g/L of exopolysaccharides (EPS), which was three-fold higher than the amount produced by H16 (10.1 g/L). EPS exhibited 59.7% of emulsification activity (EI24), higher than that of SDS and of EPS from H16 with soybean oil. To evaluate P(3HB-co-3HHx) production from soybean oil, BM3-1 was engineered with P(3HB-co-3HHx) biosynthetic genes (phaCRa, phaARe, and phaJPa). BM3-1/pPhaCJ produced 3.5 mol% of 3HHx and 37.1 g/L PHA. BM3-1/pCB81 (phaCAJ) produced 32.8 g/L PHA, including 5.9 mol% 3HHx. Physical and thermal analyses revealed that P(3HB-co-5.9 mol% 3HHx) was better than PHB. Collectively, we identified a novel strain with high vegetable oil utilization capacity for the production of EPS, with the option to engineer the strain for P(3HB-co-3HHx).

Production and characterization of a biodegradable polymer, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), using the type II methanotroph, Methylocystis sp. MJC1.

The production of polyhydroxyalkanoates (PHAs) through the biological conversion of methane is a promising solution to address both methane emissions and plastic waste. Type II methanotrophs naturally accumulate a representative PHA, poly(3-hydroxybutyrate) (PHB), using methane as the sole carbon source. In this study, we aimed to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV copolymer) with improved properties compared with PHB, using the type II methanotroph, Methylocystis sp. MJC1. We optimized the pH, valerate concentration, and valerate supply time in a one-step cultivation process using a gas bioreactor to enhance PHBV copolymer production yield and the 3-hydroxyvalerate (3HV) molar fraction. Under the optimal conditions, the biomass reached 21.3 g DCW/L, and PHBV copolymer accumulation accounted for 41.9 % of the dried cell weight, with a 3HV molar fraction of 28.4 %. The physicochemical properties of the purified PHBV copolymer were characterized using NMR, FTIR, TGA, DSC, and GPC.

Algal biochar mediated detoxification of plant biomass hydrolysate: Mechanism study and valorization into polyhydroxyalkanoates.

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.

Macroalgae as a source of sugar and detoxifier biochar for polyhydroxyalkanoates production by Halomonas sp. YLGW01 under the unsterile condition.

Macroalgae (seaweed) is considered a favorable feedstock for polyhydroxyalkanoates (PHAs) production owing to its high productivity, low land and freshwater requirement, and renewable nature. Among different microbes Halomonas sp. YLGW01 can utilize algal biomass-derived sugars (galactose and glucose) for growth and PHAs production. Biomass-derived byproducts furfural, hydroxymethylfurfural (HMF), and acetate affects Halomonas sp. YLGW01 growth and poly(3-hydroxybutyrate) (PHB) production i.e., furfural > HMF > acetate. Eucheuma spinosum biomass-derived biochar was able to remove 87.9 % of phenolic compounds from its hydrolysate without affecting sugar concentration. Halomonas sp. YLGW01 grows and accumulates a high amount of PHB at 4 % NaCl. The use of detoxified unsterilized media resulted in high biomass (6.32 ± 0.16 g cdm/L) and PHB production (3.88 ± 0.04 g/L) compared to undetoxified media (3.97 ± 0.24 g cdm/L, 2.58 ± 0.1 g/L). The finding suggests that Halomonas sp. YLGW01 has the potential to valorize macroalgal biomass into PHAs and open a new avenue for renewable bioplastic production.

Direct Monitoring of Membrane Fatty Acid Changes and Effects on the Isoleucine/Valine Pathways in an ndgR Deletion Mutant of Streptomyces coelicolor.

NdgR, a global regulator in soil-dwelling and antibiotic-producing Streptomyces, is known to regulate branched-chain amino acid metabolism by binding to the upstream region of synthetic genes. However, its numerous and complex roles are not yet fully understood. To more fully reveal the function of NdgR, phospholipid fatty acid (PLFA) analysis with gas chromatography-mass spectrometry (GC-MS) was used to assess the effects of an ndgR deletion mutant of Streptomyces coelicolor. The deletion of ndgR was found to decrease the levels of isoleucine- and leucine-related fatty acids but increase those of valine-related fatty acids. Furthermore, the defects in leucine and isoleucine metabolism caused by the deletion impaired the growth of Streptomyces at low temperatures. Supplementation of leucine and isoleucine, however, could complement this defect under cold shock condition. NdgR was thus shown to be involved in the control of branched-chain amino acids and consequently affected the membrane fatty acid composition in Streptomyces. While isoleucine and valine could be synthesized by the same enzymes (IlvB/N, IlvC, IlvD, and IlvE), ndgR deletion did not affect them in the same way. This suggests that NdgR is involved in the upper isoleucine and valine pathways, or that its control over them differs in some respect.

Lignocellulosic hydrolysate based biorefinery for marine exopolysaccharide production and application of the produced biopolymer in environmental clean-up.

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-1 and 5.32 g L-1 of EPS on supplementing 30 g L-1 glucose and 10 g L-1 xylose as the sole carbon source, respectively. Whereas on feeding Miscanthus hydrolysate (MCH) with glucose content adjusting to 20 g L-1, E. sediminis BBL-M-12 produced 6.18 g L-1 of EPS. The inhibitors study showed bacterium could tolerate higher concentrations of fermentation inhibitors include furfural (0.05%), 5-hydroxymethylfurfural (0.1%), vanillin (0.1%) and acetate (0.5%). Moreover, the EPS composition was greatly altered with the type and concentration of carbon source supplied, although ß-D-Glucopyranose, ß-D-Galactopyranose, and ß-D-Xylopyranose were the dominant monomers detected. Interestingly, E. sediminis BBL-M-12 EPS revealed excellent environmental applications like clay flocculation, oil emulsification, and removal of humic acid, textile dye, and heavy metal from the aqueous phase.

Adsorptive removal of synthetic plastic components bisphenol-A and solvent black-3 dye from single and binary solutions using pristine pinecone biochar.

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 (R2 > 0.98) and pseudo-second-order (R2 > 0.99) kinetic models, respectively. Whereas for the bisphenol-A + dye removal from binary solution, the values for bisphenol-A adsorption were best suited to Elovich (R2 > 0.98), while pseudo-second-order (R2 > 0.99) for dye removal. Similarly, the two-compartment model also demonstrated better values (R2 > 0.92) for bisphenol-A and dye removal from single and binary solutions with greater Ffast values (except for bisphenol-A in binary solution). The Langmuir isotherm model demonstrated the highest regression coefficient values (R2 > 0.99) for bisphenol-A and dye removal with the highest adsorption capacity of 38.387 mg g-1 and 346.856 mg g-1, correspondingly. Besides, the co-existence of humic acid revealed a positive impact on bisphenol-A removal, while the dye removal rate was slightly hindered in presence of humic acid. The absorption process showed monolayer coverage of biochar surface with contaminants using a chemisorption mechanism with fast reactions between functional groups on the adsorbate and adsorbent. Whereas the adsorption mechanism was primarily controlled by hydrogen bonding, hydrophobic and π-π electron-donor-acceptor interactions as confirmed by FTIR, XPS, and pH investigations.

An integrative multiomics approach to characterize anti-adipogenic and anti-lipogenic effects of Akkermansia muciniphila in adipocytes.

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.

Arctic Psychrotolerant Pseudomonas sp. B14-6 Exhibits Temperature-Dependent Susceptibility to Aminoglycosides.

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 Pseudomonas sp. B14-6 from the glacier foreland of Midtre Lovénbreen in High Arctic Svalbard. Herein, we investigated the resistance and tolerance of Pseudomonas sp. B14-6 against aminoglycosides at various temperatures. This bacterium was resistant to streptomycin and susceptible to apramycin, gentamicin, kanamycin, and tobramycin. The two putative aminoglycoside phosphotransferase genes aph1 and aph2 were the most likely contributors to streptomycin resistance. Notably, unlike the mesophilic Pseudomonas aeruginosa PA14, this cold-adapted bacterium demonstrated reduced susceptibility to all tested aminoglycosides in a temperature-dependent manner. Pseudomonas sp. B14-6 at a lower temperature formed the persister cells that shows tolerance to the 100-fold minimum inhibitory concentration (MIC) of gentamicin, as well as the partially tolerant cells that withstand 25-fold MIC gentamicin. The temperature-dependent gentamicin tolerance appears to result from reduced metabolic activity. Lastly, the partially tolerant Pseudomonas sp. B14-6 cells could slowly proliferate under the bactericidal concentrations of aminoglycosides. Our results demonstrate that Pseudomonas sp. B14-6 has a characteristic ability to form cells with a range of tolerance, which appears to be inversely proportional to its growth rate.

Bioprospecting of exopolysaccharide from marine Sphingobium yanoikuyae BBL01: Production, characterization, and metal chelation activity.

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%), ß-D-glucopyranose (21.0 ± 1.6%), α-D-mannopyranose (18.5 ± 1.2%), ß-d-mannopyranose (13.1 ± 1.4%), ß-D-xylopyranose (10.2 ± 2.1%), α-d-talopyranose (5.9 ± 1.1%), and ß-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.

Adsorptive removal of crude petroleum oil from water using floating pinewood biochar decorated with coconut oil-derived fatty acids.

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.