Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 47
Filtrar
1.
Anal Biochem ; 685: 115390, 2024 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-37951454

RESUMO

To alleviate environmental problems caused by using conventional plastics, bioplastics have garnered significant interest as alternatives to petroleum-based plastics. Despite possessing better degradability traits compared to traditional plastics, the degradation of bioplastics still demands a longer duration than initially anticipated. This necessitates the utilization of degradation strains or enzymes to enhance degradation efficiency, ensuring timely degradation. In this study, a novel screening method to identify bioplastic degraders faster was suggested to circumvent the time-consuming and laborious characteristics of solid-based plate assays. This liquid-based colorimetric method confirmed the extracellular esterase activity with p-nitrophenyl esters. It eliminated the needs to prepare plastic emulsion plates at the initial screening system, shortening the time for the overall screening process and providing more quantitative data. p-nitrophenyl hexanoate (C6) was considered the best substrate among the various p-nitrophenyl esters as substrates. The screening was performed in liquid-based 96-well plates, resulting in the discovery of a novel strain, Bacillus sp. SH09, with a similarity of 97.4% with Bacillus licheniformis. Furthermore, clear zone assays, degradation investigations, scanning electron microscopy, and gel permeation chromatography were conducted to characterize the biodegradation capabilities of the new strain, the liquid-based approach offered a swift and less labor-intensive option during the initial stages.


Assuntos
Esterases , Plásticos , Plásticos/química , Esterases/química , Ensaios de Triagem em Larga Escala , Colorimetria , Biopolímeros
2.
Anal Biochem ; 655: 114832, 2022 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-35948058

RESUMO

The increasing interest in bioplastics, with regard to future environmental issues, has rendered research on bioplastic biodegradation highly important. However, only a few tools directly monitor the degradation of bioplastics without measuring the levels of gaseous products, such as carbon dioxide. Classical nonquantitative methods, such as clear zone tests on solid plates, and less-sensitive weight-loss experiments in liquid media measured using a precision scale, are still employed to screen the microbial players associated with bioplastic degradation and monitor the biodegradation rates. However, the simultaneous monitoring of the degradation of each component of blended bioplastics has not been previously reported. In the present study, to provide information regarding the degradation rates and compositional changes of different bioplastics in a blend in a time-dependent manner, we simultaneously monitored and quantified the degradation of four bioplastics, polyhydroxybutyrate (PHB), polybutylene succinate (PBS), polycaprolactone (PCL), and poly(butylene adipate-co-terephthalate) (PBAT), by Bacillus sp. JY36 using gas chromatography-mass spectrometry (GC-MS) analysis after fatty acid methyl ester (FAME) derivatization. Our results demonstrate the feasibility of using the GC-MS-based method described here to obtain comprehensive data regarding blended bioplastics and their degradation. Moreover, our findings indicate that this method may support classical analytic tools for assessing bioplastic biodegradation.


Assuntos
Poliésteres , Biodegradação Ambiental , Cromatografia Gasosa-Espectrometria de Massas , Poliésteres/metabolismo
3.
Bioprocess Biosyst Eng ; 45(10): 1719-1729, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36121506

RESUMO

Polyhydroxybutyrate (PHB) is a bio-based, biodegradable and biocompatible plastic that has the potential to replace petroleum-based plastics. Lignocellulosic biomass is a promising feedstock for industrial fermentation to produce bioproducts such as polyhydroxybutyrate (PHB). However, the pretreatment processes of lignocellulosic biomass lead to the generation of toxic byproducts, such as furfural, 5-HMF, vanillin, and acetate, which affect microbial growth and productivity. In this study, to reduce furfural toxicity during PHB production from lignocellulosic hydrolysates, we genetically engineered Cupriavidus necator NCIMB 11599, by inserting the nicotine amide salvage pathway genes pncB and nadE to increase the NAD(P)H pool. We found that the expression of pncB was the most effective in improving tolerance to inhibitors, cell growth, PHB production and sugar consumption rate. In addition, the engineered strain harboring pncB showed higher PHB production using lignocellulosic hydrolysates than the wild-type strain. Therefore, the application of NAD salvage pathway genes improves the tolerance of Cupriavidus necator to lignocellulosic-derived inhibitors and should be used to optimize PHB production.


Assuntos
Cupriavidus necator , Petróleo , Amidas/metabolismo , Cupriavidus necator/genética , Cupriavidus necator/metabolismo , Açúcares da Dieta/metabolismo , Açúcares da Dieta/farmacologia , Furaldeído/farmacologia , Inibidores do Crescimento/metabolismo , Inibidores do Crescimento/farmacologia , Hidroxibutiratos/metabolismo , Lignina , NAD/metabolismo , NAD/farmacologia , Nicotina/metabolismo , Nicotina/farmacologia , Nitrobenzenos , Petróleo/metabolismo , Plásticos
4.
J Ind Microbiol Biotechnol ; 48(5-6)2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33956122

RESUMO

Caldicellulosiruptor bescii is the most thermophilic, cellulolytic bacterium known and has the native ability to utilize unpretreated plant biomass. Cellulase A (CelA) is the most abundant enzyme in the exoproteome of C. bescii and is primarily responsible for its cellulolytic ability. CelA contains a family 9 glycoside hydrolase and a family 48 glycoside hydrolase connected by linker regions and three carbohydrate-binding domains. A truncated version of the enzyme (TM1) containing only the endoglucanase domain is thermostable and actively degrades crystalline cellulose. A catalytically active TM1 was successfully produced via the attachment of the PelB signal peptide (P-TM1), which mediates post-translational secretion via the SecB-dependent translocation pathway. We sought to enhance the extracellular secretion of TM1 using an alternative pathway, the signal recognition particle (SRP)-dependent translocation pathway. The co-translational extracellular secretion of TM1 via the SRP pathway (D-TM1) resulted in a specific activity that was 4.9 times higher than that associated with P-TM1 overexpression. In batch fermentations, the recombinant Escherichia coli overexpressing D-TM1 produced 1.86 ± 0.06 U/ml of TM1 in the culture medium, showing a specific activity of 1.25 ± 0.05 U/mg cell, 2.7- and 3.7-fold higher than the corresponding values of the strain overexpressing P-TM1. We suggest that the TM1 secretion system developed in this study can be applied to enhance the capacity of E. coli as a microbial cell factory for the extracellular secretion of this as well as a variety proteins important for commercial production.


Assuntos
Celulase/biossíntese , Escherichia coli/metabolismo , Peptidoglicano/metabolismo , Via Secretória , Partícula de Reconhecimento de Sinal/metabolismo , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Caldicellulosiruptor/enzimologia , Caldicellulosiruptor/genética , Carboxipeptidases/genética , Celulase/genética , Celulose/metabolismo , DNA Bacteriano , Escherichia coli/genética , Fermentação , Glicosídeo Hidrolases , Microbiologia Industrial , Mutação , Peptidoglicano/genética , Domínios Proteicos , Sinais Direcionadores de Proteínas , Transporte Proteico , Proteínas Recombinantes/biossíntese
5.
Bioprocess Biosyst Eng ; 44(4): 891-899, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33486578

RESUMO

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.


Assuntos
Trifosfato de Adenosina/metabolismo , Cadaverina/química , Escherichia coli/metabolismo , Fosfato de Piridoxal/metabolismo , Saccharomyces cerevisiae , Ágar/química , Biotecnologia/métodos , Biotransformação , Cadaverina/metabolismo , Carboxiliases , Fermentação , Microbiologia Industrial/instrumentação , Microbiologia Industrial/métodos , Lisina/química , Lisina/metabolismo , Polímeros/química , Piridoxal , Regeneração
6.
Bioprocess Biosyst Eng ; 40(5): 781-789, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28204983

RESUMO

Polyhydroxyalkanoate (PHA) is a family of biodegradable polymers, and incorporation of different monomers can alter its physical properties. To produce the copolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3HV)) containing a high level of 3-hydroxyvalerate (3HV) by altering acetyl-CoA pool levels, we overexpressed an acetyl-CoA acetyltransferase (atoAD) in an engineered E. coli strain, YH090, carrying PHA synthetic genes bktB, phaB, and phaC. It was found that, with introduction of atoAD and with propionate as a co-substrate, 3HV fraction in PHA was increased up to 7.3-fold higher than a strain without atoAD expressed in trans (67.9 mol%). By the analysis of CoA pool concentrations in vivo and in vitro using HPLC and LC-MS, overexpression of AtoAD was shown to decrease the amount of acetyl-CoA and increase the propionyl-CoA/acetyl-CoA ratio, ultimately resulting in an increased 3HV fraction in PHA. Finally, synthesis of P(3HB-co-3HV) containing 57.9 mol% of 3HV was achieved by fed-batch fermentation of YJ101 with propionate.


Assuntos
Acetil-CoA C-Acetiltransferase/biossíntese , Proteínas de Escherichia coli/biossíntese , Escherichia coli/metabolismo , Ácidos Pentanoicos/metabolismo , Poliésteres/metabolismo , Acetil-CoA C-Acetiltransferase/genética , Escherichia coli/genética , Proteínas de Escherichia coli/genética
7.
Bioprocess Biosyst Eng ; 38(8): 1479-84, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25820819

RESUMO

Every year, the amount of chemosynthetic plastic accumulating in the environment is increasing, and significant time is required for decomposition. Bio-based, biodegradable plastic is a promising alternative, but its production is not yet a cost effective process. Decreasing the production cost of polyhydroxyalkanoate by utilizing renewable carbon sources for biosynthesis is an important aspect of commercializing this biodegradable polymer. An Escherichia coli strain that expresses a functional amylase and accumulate polyhydroxybutyrate (PHB), was constructed using different plasmids containing the amylase gene of Panibacillus sp. and PHB synthesis genes from Ralstonia eutropha. This engineered strain can utilize starch as the sole carbon source. The maximum PHB production (1.24 g/L) was obtained with 2% (w/v) starch in M9 media containing 0.15% (w/v) yeast extract and 10 mM glycine betaine. The engineered E. coli SKB99 strain can accumulate intracellular PHB up to 57.4% of cell dry mass.


Assuntos
Escherichia coli/metabolismo , Hidroxibutiratos/metabolismo , Engenharia Metabólica , Poliésteres/metabolismo , Amido/metabolismo , Amilases/biossíntese , Cupriavidus necator/enzimologia , Cupriavidus necator/genética , Escherichia coli/genética , Proteínas Recombinantes/biossíntese
8.
Sci Total Environ ; 914: 169926, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38199349

RESUMO

Microplastics (MPs) pollution has emerged as a critical environmental issue with far-reaching consequences for ecosystems and human health. These are plastic particles measuring <5 mm and are categorized as primary and secondary based on their origin. Primary MPs are used in various products like cosmetics, scrubs, body wash, and toothpaste, while secondary MPs are generated through the degradation of plastic products. These have been detected in seas, rivers, snow, indoor air, and seafood, posing potential risks to human health through the food chain. Detecting and quantifying MPs are essential to understand their distribution and abundance in the environment. Various microscopic (fluorescence microscopy, scanning electron microscopy) and spectroscopy techniques (FTIR, Raman spectroscopy, X-ray photoelectron spectroscopy) have been reported to analyse MPs. Despite the challenges in scalable removal methods, biological systems have emerged as promising options for eco-friendly MPs remediation. Algae, bacteria, and fungi have shown the potential to adsorb and degrade MPs in wastewater treatment plants (WWTPs) offering hope for mitigating this global crisis. This review examines the sources, impacts, detection, and biological removal of MPs, highlighting future directions in this crucial field of environmental conservation. By fostering global collaboration and innovative research a path towards a cleaner and healthier planet for future generations can be promised.


Assuntos
Microplásticos , Poluentes Químicos da Água , Humanos , Plásticos/análise , Águas Residuárias , Ecossistema , Poluentes Químicos da Água/análise , Monitoramento Ambiental
9.
Int J Biol Macromol ; 254(Pt 1): 127475, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37863147

RESUMO

Polyhydroxybutyrate (PHB) is a well-known biodegradable bioplastic synthesized by microorganisms and can be produced from volatile fatty acids (VFAs). Among VFAs acetate can be utilized by Halomonas sp. YLGW01 for growth and PHB production. In this study, Halomonas sp. JJY01 was developed through introducing acetyl-CoA acetyltransferase (atoAD) with LacIq-Ptrc promoter into Halomonas sp. YLGW01. The effect of expression of atoAD on acetate was investigated by comparison with acetate consumption and PHB production. Shake-flask study showed that Halomonas sp. JJY01 increased acetate consumption rate, PHB yield and PHB production (0.27 g/L/h, 0.075 g/g, 0.72 g/L) compared to the wild type strain (0.17 g/L/h, 0.016 g/g, 0.11 g/L). In 10 L fermenter scale fed-batch fermentation, the growth of Halomonas sp. JJY01 resulted in higher acetate consumption rate, PHB yield and PHB titer (0.55 g/L/h, 0.091 g/g, 4.6 g/L) than wild type strain (0.35 g/L/h, 0.067 h/h, 2.9 g/L). These findings demonstrate enhanced acetate utilization and PHB production through the introduction of atoAD in Halomonas strains.


Assuntos
Halomonas , Hidroxibutiratos , Hidroxibutiratos/metabolismo , Halomonas/genética , Halomonas/metabolismo , Acetil-CoA C-Acetiltransferase/metabolismo , Poli-Hidroxibutiratos , Acetatos/metabolismo , Poliésteres/metabolismo
10.
Int J Biol Macromol ; 257(Pt 2): 128687, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38101655

RESUMO

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.


Assuntos
Poli-Hidroxialcanoatos , Biopolímeros , Ácido 3-Hidroxibutírico , Hidroxibutiratos , Solventes
11.
J Microbiol Biotechnol ; 34(7): 1530-1543, 2024 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-38973389

RESUMO

With an increase in the commercialization of bioplastics, the importance of screening for plastic-degrading strains and microbes has emerged. Conventional methods for screening such strains are time-consuming and labor-intensive. Therefore, we suggest a method for quickly and effectively screening plastic-degrading microbial strains through dual esterase assays for soil and isolated strains, using p-nitrophenyl alkanoates as substrates. To select microbe-abundant soil, the total amount of phospholipid fatty acids (PLFAs) included in each soil sample was analyzed, and esterase assays were performed for each soil sample to compare the esterase activity of each soil. In addition, by analyzing the correlation coefficients and sensitivity between the amount of PLFAs and the degree of esterase activity according to the substrate, it was confirmed that substrate pNP-C2 is the most useful index for soil containing several microbes having esterase activity. In addition, esterase assays of the isolated strains allowed us to select the most active strain as the degrading strain, and 16S rRNA results confirmed that it was Bacillus sp. N04 showed the highest degradation activity for polybutylene succinate (PBS) as measured in liquid culture for 7 days, with a degradation yield of 99%. Furthermore, Bacillus sp. N04 showed degradation activity against various bioplastics. We propose the dual application of p-nitrophenyl alkanoates as an efficient method to first select the appropriate soil and then to screen for plastic-degrading strains in it, and conclude that pNP-C2 in particular, is a useful indicator.


Assuntos
Biodegradação Ambiental , Esterases , Nitrofenóis , Microbiologia do Solo , Nitrofenóis/metabolismo , Esterases/metabolismo , Solo/química , Bactérias/metabolismo , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/classificação , RNA Ribossômico 16S/genética , Ácidos Graxos/metabolismo , Bacillus/metabolismo , Bacillus/genética , Bacillus/isolamento & purificação , Fosfolipídeos/metabolismo , Plásticos Biodegradáveis/metabolismo
12.
Int J Biol Macromol ; 266(Pt 2): 131332, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38574905

RESUMO

Polyhydroxyalkanoates (PHAs) are promising alternatives to existing petrochemical-based plastics because of their bio-degradable properties. However, the limited structural diversity of PHAs has hindered their application. In this study, high mole-fractions of Poly (39 mol% 3HB-co-17 mol% 3 HV-co-44 mol% 4 HV) and Poly (25 mol% 3HB-co-75 mol% 5 HV) were produced from 4- hydroxyvaleric acid and 5-hydroxyvaleric acid, using Cupriavidus necator PHB-4 harboring the gene phaCBP-M-CPF4 with modified sequences. In addition, the complex toxicity of precursor mixtures was tested, and it was confirmed that the engineered C. necator was capable of synthesizing Poly (32 mol% 3HB-co-11 mol% 3 HV-co-25 mol% 4 HV-co-32 mol% 5 HV) at low mixture concentrations. Correlation analyses of the precursor ratio and the monomeric mole fractions indicated that each mole fractions could be precisely controlled using the precursor proportion. Physical property analysis confirmed that Poly (3HB-co-3 HV-co-4 HV) is a rubber-like amorphous polymer and Poly (3HB-co-5 HV) has a high tensile strength and elongation at break. Poly (3HB-co-3 HV-co-4 HV-co-5 HV) had a much lower glass transition temperature than the co-, terpolymers containing 3 HV, 4 HV and 5 HV. This study expands the range of possible physical properties of PHAs and contributes to the realization of custom PHA production by suggesting a method for producing PHAs with various physical properties through mole-fraction control of 3 HV, 4 HV and 5 HV.


Assuntos
Cupriavidus necator , Poli-Hidroxialcanoatos , Cupriavidus necator/metabolismo , Cupriavidus necator/genética , Poli-Hidroxialcanoatos/biossíntese , Poli-Hidroxialcanoatos/química , Ácido 3-Hidroxibutírico/química , Ácido 3-Hidroxibutírico/biossíntese , Ácidos Pentanoicos/metabolismo , Ácidos Pentanoicos/química , Poliésteres/química , Poliésteres/metabolismo
13.
Bioresour Technol ; 389: 129853, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37813313

RESUMO

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.


Assuntos
Methylocystaceae , Poliésteres , Hidroxibutiratos , Valeratos , Metano
14.
Bioresour Technol ; 368: 128332, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36414137

RESUMO

Hydrogen has gained attention as an alternative source of energy because of its non-polluting nature as on combustion it produces only water. Biological methods are eco-friendly and have benefits in waste management and hydrogen production simultaneously. The use of algal biomass as feedstock in dark fermentation is advantageous because of its low lignin content, high growth rate, and carbon-fixation ability. The major bottlenecks in biohydrogen production are its low productivity and high production costs. To overcome these issues, many advances in the area of biomass pretreatment to increase sugar release, understanding of algal biomass composition, and development of fermentation strategies for the complete recovery of nutrients are ongoing. Recently, mixed substrate fermentation, multistep fermentation, and the use of nanocatalysts to improve hydrogen production have increased. This review article evaluates the current progress in algal biomass pretreatment, key factors, and possible solutions for increasing hydrogen production.


Assuntos
Hidrogênio , Lignina , Biomassa , Fermentação , Nutrientes
15.
Int J Biol Macromol ; 244: 125298, 2023 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-37315675

RESUMO

The use of bioplastics, which can alleviate environmental pollution caused by non-degradable bioplastics, has received attention. As there are many types of bioplastics, method that can treat them simultaneously is important. Therefore, Bacillus sp. JY35 which can degrade different types of bioplastics, was screened in previous study. Most types of bioplastics, such as polyhydroxybutyrate (PHB), (P(3HB-co-4HB)), poly(butylene adipate-co-terephthalate) (PBAT), polybutylene succinate (PBS), and polycaprolactone (PCL), can be degraded by esterase family enzymes. To identify the genes that are involved in bioplastic degradation, analysis with whole-genome sequencing was performed. Among the many esterase enzymes, three carboxylesterase and one triacylglycerol lipase were identified and selected based on previous studies. Esterase activity using p-nitrophenyl substrates was measured, and the supernatant of JY35_02679 showed strong emulsion clarification activity compared with others. In addition, when recombinant E. coli was applied to the clear zone test, only the JY35_02679 gene showed activity in the clear zone test with bioplastic containing solid cultures. Further quantitative analysis showed 100 % PCL degradation at 7 days and 45.7 % PBS degradation at 10 days. We identified a gene encoding a bioplastic-degrading enzyme in Bacillus sp. JY35 and successfully expressed the gene in heterologous E. coli, which secreted esterases with broad specificity.


Assuntos
Bacillus , Bacillus/genética , Escherichia coli , Biopolímeros , Esterases/genética
16.
Int J Biol Macromol ; 225: 757-766, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36400208

RESUMO

Phasin is a surface-binding protein of polyhydroxyalkanoate (PHA) granules that is encoded by the phaP gene. As its expression increases, PHA granules become smaller, to increase their surface area, and are densely packed inside the cell, thereby increasing the PHA content. A wide range of PHA-producing bacteria have phaP genes; however, their PHA productivity differs, although they are derived from the cognate bacterial host cell. Modulating phasin expression could be a new strategy to enhance PHA production. This study aimed to characterize the effect of heterologous phasins on the reconstitution of E. coli BL21(DE3) and determine the best synergistic phaP gene combination to produce polyhydroxybutyrate (PHB). We identified novel phasins from a PHB high-producer strain, Halomonas sp. YLGW01, and introduced a combination of phaP genes into Escherichia coli. The resulting E. coli phaP1,3 strain had enhanced PHB production by 2.9-fold, leading to increased cell mass and increased PHB content from 48 % to 65 %. This strain also showed increased tolerance to inhibitors, such as furfural and vanillin, enabling the utilization of lignocellulose biosugar as a carbon source. These results suggested that the combination of phaP1 and phaP3 genes from H. sp. YLGW01 could increase PHB production and robustness.


Assuntos
Escherichia coli , Lectinas de Plantas , Escherichia coli/genética , Escherichia coli/metabolismo , Lectinas de Plantas/metabolismo , Proteínas de Bactérias/química , Hidroxibutiratos/metabolismo , Poliésteres/metabolismo
17.
J Microbiol Biotechnol ; 33(8): 1076-1083, 2023 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-37311705

RESUMO

Poly(3-hydroxybutyrate) (PHB) is a biodegradable and biocompatible bioplastic. Effective PHB degradation in nutrient-poor environments is required for industrial and practical applications of PHB. To screen for PHB-degrading strains, PHB double-layer plates were prepared and three new Bacillus infantis species with PHB-degrading ability were isolated from the soil. In addition, phaZ and bdhA of all isolated B. infantis were confirmed using a Bacillus sp. universal primer set and established polymerase chain reaction conditions. To evaluate the effective PHB degradation ability under nutrient-deficient conditions, PHB film degradation was performed in mineral medium, resulting in a PHB degradation rate of 98.71% for B. infantis PD3, which was confirmed in 5 d. Physical changes in the degraded PHB films were analyzed. The decrease in molecular weight due to biodegradation was confirmed using gel permeation chromatography and surface erosion of the PHB film was observed using scanning electron microscopy. To the best of our knowledge, this is the first study on B. infantis showing its excellent PHB degradation ability and is expected to contribute to PHB commercialization and industrial composting.


Assuntos
Bacillus , Solo , Ácido 3-Hidroxibutírico , Hidroxibutiratos/metabolismo , Poliésteres/metabolismo , Bacillus/genética , Bacillus/metabolismo , Hidrolases de Éster Carboxílico/metabolismo
18.
Int J Biol Macromol ; 236: 123997, 2023 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-36907298

RESUMO

Petrochemical-based plastics are hardly biodegradable and a major cause of environmental pollution, and polyhydroxybutyrate (PHB) is attracting attention as an alternative due to its similar properties. However, the cost of PHB production is high and is considered the greatest challenge for its industrialization. Here, crude glycerol was used as a carbon source for more efficient PHB production. Among the 18 strains investigated, Halomonas taeanenisis YLGW01 was selected for PHB production due to its salt tolerance and high glycerol consumption rate. Furthermore, this strain can produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (P(3HB-co-3 HV)) with 17 % 3 HV mol fraction when a precursor is added. PHB production was maximized through medium optimization and activated carbon treatment of crude glycerol, resulting in 10.5 g/L of PHB with 60 % PHB content in fed-batch fermentation. Physical properties of the produced PHB were analyzed, i.e., weight average molecular weight (6.8 × 105), number average molecular weight (4.4 × 105), and the polydispersity index (1.53). In the universal testing machine analysis, the extracted intracellular PHB showed a decrease in Young's modulus, an increase in Elongation at break, greater flexibility than authentic film, and decreased brittleness. This study confirmed that YLGW01 is a promising strain for industrial PHB production using crude glycerol.


Assuntos
Glicerol , Halomonas , Poliésteres , Plásticos , Hidroxibutiratos
19.
Bioresour Technol ; 359: 127499, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35718248

RESUMO

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.


Assuntos
Carbono , Lignina , Fermentação , Glucose
20.
Int J Biol Macromol ; 208: 809-818, 2022 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-35364206

RESUMO

Polyhydroxybutyrate (PHB) is a potential substitute for plastics derived from fossil fuels, owing to its biodegradable and biocompatible properties. Lignocellulosic biomass could be used to reduce PHB production costs; however, the co-utilization of sugars, such as glucose and xylose, without catabolite repression is a difficult problem to be solved. Here, we selected a novel Loktanella sp. SM43 from a marine environment and optimized the conditions for PHB production. Loktanella sp. SM43 showed high PHB production (66.5% content) from glucose. When glucose and xylose were used together, this strain showed high utilization of both substrates compared to other high PHB-producers such as Halomonas sp. and Cupriavidus necator, which showed glucose preference. Loktanella sp. SM43 showed high growth and PHB production with lignocellulosic hydrolysates. When pine tree hydrolysates were used, PHB production was the highest at 3.66 ± 0.01 g/L, followed by Miscanthus (3.46 ± 0.09 g/L) and barley straw hydrolysate (3.36 ± 0.36 g/L). Overall, these results reveal the potential of Loktanella sp. SM43 to produce PHB using various lignocellulosic hydrolysates as feedstock and the first systematic study for PHB production with Loktanella sp. The approach of screening novel strains is a strategy to overcome co-utilization of sugars without genetic engineering.


Assuntos
Glucose , Xilose , Biomassa , Fermentação , Hidroxibutiratos/química , Lignina , Açúcares
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA