Enzyme and Microbial Technology最新文献_第5页

Protein engineering of an alkaline protease from Bacillus licheniformis (BLAP) for efficient and specific chiral resolution of the racemic ethyl tetrahydrofuroate 地衣芽孢杆菌碱性蛋白酶（BLAP）的蛋白质工程，用于高效、特异性手性解析外消旋四氢糠酸乙酯。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-10-03 DOI: 10.1016/j.enzmictec.2024.110523

Xinjun Yu , Yichao Li , Zhaoxia Qian, Litian Wei, Jing Xie, Meijun Tong, Yinjun Zhang

{"title":"Protein engineering of an alkaline protease from Bacillus licheniformis (BLAP) for efficient and specific chiral resolution of the racemic ethyl tetrahydrofuroate","authors":"Xinjun Yu , Yichao Li , Zhaoxia Qian, Litian Wei, Jing Xie, Meijun Tong, Yinjun Zhang","doi":"10.1016/j.enzmictec.2024.110523","DOIUrl":"10.1016/j.enzmictec.2024.110523","url":null,"abstract":"<div><div>Enzymatic resolution of ethyl tetrahydrofuroate to produce (<em>S</em>)-2-ethyl tetrahydrofuroate and (<em>R</em>)-2-tetrahydrofuroic acid is a green biomanufacturing strategy. However, enzymatic activity and selectivity are still limiting factors of their industrial applications and development. In previous study, we incidentally found that a <em>Bacillus licheniformis</em> alkaline protease (BLAP), not a lipase, could specifically resolve ethyl tetrahydrofuroate to produce (<em>S</em>)-2-ethyl tetrahydrofuroate and (<em>R</em>)-2-tetrahydrofuroic acid. In this study, the point-saturation-mutation libraries based on the seven amino acid sites (L105, I113, P114, L115, V309, Y310, and M326) were constructed and screened using the molecular docking technology. It was found that activity of the mutant BLAP<sup>Y310E</sup> reached 182.78 U/mL with high stereoselectivity, 3.14 times higher than that of the wild-type BLAP. Further simulated mutation analysis showed that the Y310E mutation increased the distance from the substrate ligand to the binding pocket from 2.3 Å to 4.5 Å, reducing steric hindrance to the active center. Under the optimal conditions and after 3.5 h of reaction catalyzed by BLAP<sup>Y310E</sup>, 200 mM ethyl tetrahydrofuroate was converted to (<em>S</em>)-2-ethyl tetrahydrofuroate and (<em>R</em>)-2-tetrahydrofuroic acid with the <em>ee</em> values of 99.9 % and 68.63 %, respectively. The enantiomeric ratio of BLAP<sup>Y310E</sup> was 105.5, which was 30.23 times higher than that of BLAP. This study advances the comprehension of protease activity and selectivity mechanisms in resolving ester substances and lays a robust foundation for the industrial production of the optically pure (<em>S</em>)-2-ethyl tetrahydrofuroate and (<em>R</em>)-2-tetrahydrofuroic acid via biological enzymatic methods.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110523"},"PeriodicalIF":3.4,"publicationDate":"2024-10-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142388977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterization of lycopene β-cyclase from Dunaliella bardawil for enhanced β-carotene production and salt tolerance 巴达维杜莎藻番茄红素β-环化酶的表征，以提高β-胡萝卜素产量和耐盐性。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-10-01 DOI: 10.1016/j.enzmictec.2024.110520

Yu-Chen Xie , Zhi-Wei Ye , Jv-Liang Dai , Hao-Hong Chen , Jian-Guo Jiang

{"title":"Characterization of lycopene β-cyclase from Dunaliella bardawil for enhanced β-carotene production and salt tolerance","authors":"Yu-Chen Xie , Zhi-Wei Ye , Jv-Liang Dai , Hao-Hong Chen , Jian-Guo Jiang","doi":"10.1016/j.enzmictec.2024.110520","DOIUrl":"10.1016/j.enzmictec.2024.110520","url":null,"abstract":"<div><div><em>Dunaliella</em> can accumulate more β-carotene (10 % or even more of the dry weight of cells) than any other species. Lycopene β-cyclase (LcyB) is the key enzyme in the catalysis of lycopene to β-carotene. In the present research, we used <em>Escherichia coli</em> BL21 (DE3) as host to construct two different types of engineering bacteria, one expressing the <em>D. bardawil</em> LcyB and the other expressing the orthologue <em>Erwinia uredovora</em> crtY. The catalytic ability of LcyB and CrtY were evaluated by comparing the β-carotene yields of the two <em>E. coli</em> BL21(DE3) strains, whose salt tolerance was simultaneously compared by cultivated them under different NaCl concentrations (1 %, 2 %, and 4 %). We also interfered with the <em>LcyB</em> gene to investigate the effect of <em>LcyB</em> in <em>D. bardawil</em>. Results displayed that the β-carotene yield of the LcyB-transformant significantly increased by about 48 % compared with the crtY-transformant. Additionally, <em>LcyB</em> was verified to be able to enhance the salt tolerance of <em>E. coli</em> BL21 (DE3). It is concluded that <em>D. bardawil</em> LcyB not only has better catalytic ability but also is able to confer salt tolerance to cells. Interfering <em>D. bardawil LcyB</em> induced the low expression of LcyB and the changes of growth and carotenoids metabolism in <em>D. bardawil.</em></div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110520"},"PeriodicalIF":3.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142388976","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Action pattern of Sulfolobus O-α-glycoligase for synthesis of highly water soluble resveratrol 3,4′-α-diglucoside 硫醇杆菌 O-α-糖苷酶合成高水溶性白藜芦醇 3,4′-α- 二糖苷的作用模式

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-09-24 DOI: 10.1016/j.enzmictec.2024.110518

Hee-Won Ahn , Jetendra Kumar Roy , Jaeick Lee , Mi-Jin Lee , Sang-Ho Yoo , Young-Wan Kim

{"title":"Action pattern of Sulfolobus O-α-glycoligase for synthesis of highly water soluble resveratrol 3,4′-α-diglucoside","authors":"Hee-Won Ahn , Jetendra Kumar Roy , Jaeick Lee , Mi-Jin Lee , Sang-Ho Yoo , Young-Wan Kim","doi":"10.1016/j.enzmictec.2024.110518","DOIUrl":"10.1016/j.enzmictec.2024.110518","url":null,"abstract":"<div><div>This study presents the enzymatic synthesis of resveratrol-3,4′-O-α-diglucoside (RDG) using a hyperactive O-α-glycoligase (MalA-D416R/Q450S) and α-glucopyranosyl fluoride as the donor substrate. The transglycosylation rate for resveratrol by MalA-D416R/Q450S was maximized in 100 mM Tris-HCl (pH 9.5) containing 20 % DMSO at 45°C. Because the p<em>K</em><sub>a</sub> of the 4′-OH group of resveratrol is lower than that of the 3-OH group, the 4′-OH group is more nucleophilic at the alkaline pH, leading to a preference for glycosylation at the 4′-OH site rather than the 3-OH site. This preference makes resveratrol 3-O-α-glucoside (R3G) as the more efficient acceptor than resveratrol 4′-O-α-glucoside (R4′G), resulting in negligible production of resveratrol 3-O-α-glucoside (R3G) due to its complete consumption in the second transglycosylation reaction when using a 2:1 ratio of donor to acceptor substrates. From a preparative scale reaction, R4′G and RDG were isolated with yields of 41.2 % and 43.3 %, respectively. The water solubility of RDG exceeded 1.67 M, which represents more than a 9,800-fold improvement compared to resveratrol. In a hydrolysis experiment using intestinal α-glycosidase from rat, the α-glucosides of resveratrol (R4′G and RDG) were completely deglycosylated to the aglycone.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110518"},"PeriodicalIF":3.4,"publicationDate":"2024-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142328131","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization 酶固定化促进不同底物蛋白改变菲辛的特异性

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-09-21 DOI: 10.1016/j.enzmictec.2024.110517

Alex D. Gonzalez-Vasquez , El Siar Hocine , Marcela Urzúa , Javier Rocha-Martin , Roberto Fernandez-Lafuente

{"title":"Changes in ficin specificity by different substrate proteins promoted by enzyme immobilization","authors":"Alex D. Gonzalez-Vasquez , El Siar Hocine , Marcela Urzúa , Javier Rocha-Martin , Roberto Fernandez-Lafuente","doi":"10.1016/j.enzmictec.2024.110517","DOIUrl":"10.1016/j.enzmictec.2024.110517","url":null,"abstract":"<div><div>Ficin extract has been immobilized using different supports: glyoxyl and Aspartic/1,6 hexamethylenediamine (Asp/HA) agarose beads. The latter was later submitted to glutaraldehyde modification to get covalent immobilization. The activities of these 3 kinds of biocatalysts were compared utilizing 4 different substrates, casein, hemoglobin and bovine serum albumin and benzoyl-arginine-p-nitroanilide at pH 7 and 5. Using glyoxyl-agarose, the effect of enzyme-support reaction time on the activity versus the four substrates at both pH values was studied. Reaction time has been shown to distort the enzyme due to an increase in the number of covalent support-enzyme bonds. Surprisingly, for all the substrates and conditions the prolongation of the enzyme-support reaction did not imply a decrease in enzyme activity. Using the Asp/HA supports (with different amount of HA) differences in the effect on enzyme activity versus the different substrates are much more significant, while with some substrates the immobilization produced a decrease in enzyme activity, with in other cases the activity increased. These different effects are even increased after glutaraldehyde treatment. That way, the conformational changes induced by the biocatalyst immobilization or the chemical modification fully altered the enzyme protein specificity. This may also have some implications when following enzyme inactivation.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110517"},"PeriodicalIF":3.4,"publicationDate":"2024-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0141022924001248/pdfft?md5=4a8c2db0aa168bda301afda3b77f7953&pid=1-s2.0-S0141022924001248-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142312640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Poly(3-hydroxybutyrate) production for food packaging from biomass derived carbohydrates by cupriavidus necator DSM 545 利用坏死葡萄球菌 DSM 545 从生物质中提取的碳水化合物生产用于食品包装的聚（3-羟基丁酸）乙酸酯

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-09-17 DOI: 10.1016/j.enzmictec.2024.110516

Gianfrancesco Russo , Paola Scocca , Mattia Gelosia , Giacomo Fabbrizi , Tommaso Giannoni , Stefania Urbani , Sonia Esposto , Andrea Nicolini

{"title":"Poly(3-hydroxybutyrate) production for food packaging from biomass derived carbohydrates by cupriavidus necator DSM 545","authors":"Gianfrancesco Russo , Paola Scocca , Mattia Gelosia , Giacomo Fabbrizi , Tommaso Giannoni , Stefania Urbani , Sonia Esposto , Andrea Nicolini","doi":"10.1016/j.enzmictec.2024.110516","DOIUrl":"10.1016/j.enzmictec.2024.110516","url":null,"abstract":"<div><p>The extensive utilization of conventional plastics has resulted in a concerning surge in waste. A potential solution lies in biodegradable polymers mostly derived from renewable sources. <em>Cupriavidus necator</em> DSM 545 is a microorganism capable, under stress conditions, of intracellularly accumulating Poly(3-hydroxybutyrate) (PHB), a bio-polyester. This study aimed to identify optimal conditions to maximize the intracellular accumulation of PHB and its global production using natural media obtained by processing lignocellulosic residues of cardoon, a low-cost feedstock. An intracellular PHB accumulation was observed in all of the tested media, indicating a metabolic stress induced by the lack of macronutrients. Increasing C/N ratios led to a significant decrease in cellular biomass and PHB production. Furthermore <em>C. necator</em> DSM 545 was incapable of consuming more than 25 g/L of supplied monosaccharides. Surprisingly, in the samples supplied with 60 % of the pentose-rich liquid fraction, complete consumption of xylose was observed. This result was also confirmed by subsequent tests using Medium 1 growth media containing xylose as the sole carbon source. Using a diluted medium with a C/N ratio of 5, a PHB production of 5.84 g/L and intracellular PHB accumulation of 77 % w/w were respectively achieved. Finally, comparative shelf-life tests conducted against conventional pre-packaging materials in PP suggested that PHB films performed similarly in preserve ready-to-eat products.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110516"},"PeriodicalIF":3.4,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0141022924001236/pdfft?md5=c85d42688356aef8de5e917227dcb749&pid=1-s2.0-S0141022924001236-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142271641","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Characterization of Runella zeae D-mannose 2-epimerase and its expression in Bacillus subtilis for D-mannose production from D-glucose Runella zeae D-甘露糖 2-酰亚胺酶的特征及其在枯草芽孢杆菌中的表达，以利用 D-葡萄糖生产 D-甘露糖

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-09-05 DOI: 10.1016/j.enzmictec.2024.110506

Yuhan Wei , Wei Xu , Wenli Zhang , Penka Petrova , Kaloyan Petrov , Dawei Ni , Wanmeng Mu

{"title":"Characterization of Runella zeae D-mannose 2-epimerase and its expression in Bacillus subtilis for D-mannose production from D-glucose","authors":"Yuhan Wei , Wei Xu , Wenli Zhang , Penka Petrova , Kaloyan Petrov , Dawei Ni , Wanmeng Mu","doi":"10.1016/j.enzmictec.2024.110506","DOIUrl":"10.1016/j.enzmictec.2024.110506","url":null,"abstract":"<div><p>D-Mannose 2-epimerase (MEase) catalyzes the bioconversion between D-glucose and D-mannose. It is an important potential biocatalyst for large-scale production of D-mannose, a functional monosaccharide used in pharmaceutical and food industries. In this study, a new microbial MEase was characterized from <em>Runella zeae</em> DSM 19591. The enzyme was purified by one-step nickel-affinity chromatography and determined to be a dimeric protein with two identical subunits of approximately 86.1 kDa by gel filtration. The enzyme showed the highest activity at pH 8.0 and 40 °C, with a specific activity of 2.99 U/mg on D-glucose and 3.71 U/mg on D-mannose. The melting temperature (<em>T</em><sub>m</sub>) was 49.4 °C and the half-life was 115.14 and 3.23 h at 35 and 40 °C, respectively. The purified enzyme (1 U/mL) produced 115.7 g/L of D-mannose from 500 g/L of D-glucose for 48 h, with a conversion ratio of 23.14 %. It was successfully expressed in <em>Bacillus subtilis</em> WB600 via pP43NMK as the vector. The highest fermentation activity was 10.58 U/mL after fed-batch cultivation for 28 h, and the whole cells of recombinant <em>B. subtilis</em> produced 114.0 g/L of D-mannose from 500 g/L of D-glucose, with a conversion ratio of 22.8 %.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110506"},"PeriodicalIF":3.4,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142167248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Penicillin binding proteins-based immunoassay for the selective and quantitative determination of beta-lactam antibiotics 基于青霉素结合蛋白的免疫测定法，用于选择性定量测定β-内酰胺类抗生素。

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-09-04 DOI: 10.1016/j.enzmictec.2024.110507

Rilong Liu , Hangzhen Lan , Song Yan , Lu Huang , Daodong Pan , Yichun Wu

{"title":"Penicillin binding proteins-based immunoassay for the selective and quantitative determination of beta-lactam antibiotics","authors":"Rilong Liu , Hangzhen Lan , Song Yan , Lu Huang , Daodong Pan , Yichun Wu","doi":"10.1016/j.enzmictec.2024.110507","DOIUrl":"10.1016/j.enzmictec.2024.110507","url":null,"abstract":"<div><p>An immunoassay method based on penicillin-binding protein (PBP) was developed for the quantitative determination of 10 kinds of beta-lactam antibiotics (BLAs). First, two kinds of PBPs, which are named PBP1a and PBP2x, were expressed and purified, and they were characterized by SDS-PAGE and western blotting analysis. Then, the binding activity of PBP1a and PBP2x to template BLAs, cefquinome (CEFQ) and ampicillin (AMP), was determined. The effect of the buffer solution system, e.g., pH, ion concentration, and organic solvent, on the immune interaction efficiency between PBPs and BLAs was also evaluated. In the end, the PBP-based immunoassay method was developed and validated for the detection of 10 kinds of BLAs. Under optimal conditions, PBPs exhibited high binding affinity to BLAs. In addition, this method showed a high sensitivity for the detection of 10 kinds of BLAs with the limits of detection from 0.21 to 9.12 ng/mL, which are much lower than their corresponding maximum residual limit of European Union (4–100 ng/mL). Moreover, the developed PBP-immunoassay was employed for BLA detection from milk samples, and satisfactory recoveries (68.9–101.3 %) were obtained.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"181 ","pages":"Article 110507"},"PeriodicalIF":3.4,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142145407","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mutant β-fructofuranosidase synthesizing blastose [β-d-Fruf-(2→6)-d-Glcp] 合成爆炸糖[β-d-Fruf-(2→6)-d-Glcp]的突变体β-呋喃果糖苷酶

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-08-25 DOI: 10.1016/j.enzmictec.2024.110500

Atsuki Takagi, Takayoshi Tagami, Masayuki Okuyama

{"title":"Mutant β-fructofuranosidase synthesizing blastose [β-d-Fruf-(2→6)-d-Glcp]","authors":"Atsuki Takagi, Takayoshi Tagami, Masayuki Okuyama","doi":"10.1016/j.enzmictec.2024.110500","DOIUrl":"10.1016/j.enzmictec.2024.110500","url":null,"abstract":"<div><p>Fructooligosaccharides (FOS) are leading prebiotics that help keep the gut healthy and aid wellness by stimulating the growth and activity of beneficial intestinal bacteria. The best-studied FOS are inulin-type FOS, mainly oligosaccharides with β-Fru<em>f</em>-(2→1)-Fru<em>f</em> linkages, including 1-kestose [β-Fru<em>f</em>-(2→1)-β-Fru<em>f</em>-(2↔1)-α-Glc<em>p</em>] and nystose [β-Fru<em>f</em>-(2→1)-β-Fru<em>f</em>-(2→1)-β-Fru<em>f</em>-(2↔1)-α-Glc<em>p</em>]. However, the properties of other types of FOS—levan-type FOS with β-Fru<em>f</em>-(2→6)-Fru<em>f</em> linkages and neo-type FOS with β-Fru<em>f</em>-(2→6)-Glc<em>p</em> linkages—remain ambiguous because efficient methods have not been established for their synthesis. Here, using site-saturation mutation of residue His79 of β-fructofuranosidase from <em>Zymomonas mobilis</em> NBRC13756, we successfully obtained a mutant β-fructofuranosidase that specifically produces neo-type FOS. The H79G enzyme variant loses the native β-Fru<em>f</em>-(2→1)-Fru-transfer ability (which produces 1-kestose), and instead has β-Fru<em>f</em>-(2→6)-Glc-transfer ability and produces neokestose. Its hydrolytic activity specific to the β-Fru<em>f</em>-(2↔1)-α-Glc<em>p</em> bond of neokestose then yields blastose [β-Fru<em>f</em>-(2→6)-Glc<em>p</em>]. The enzyme produces 0.4 M blastose from 1.0 M sucrose (80 % of the theoretical yield). The production system for blastose established here will contribute to the elucidation of the physiological functions of this disaccharide.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110500"},"PeriodicalIF":3.4,"publicationDate":"2024-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142058103","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Semi-rational engineering of ω-transaminase for enhanced enzymatic activity to 2-ketobutyrate 对ω-反转氨酶进行半合理工程设计，以增强其对 2-酮丁酸的酶活性

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-08-24 DOI: 10.1016/j.enzmictec.2024.110505

Lili Zhang , Yu Hong , Jiapeng Lu , Yi Wang , Wei Luo

{"title":"Semi-rational engineering of ω-transaminase for enhanced enzymatic activity to 2-ketobutyrate","authors":"Lili Zhang , Yu Hong , Jiapeng Lu , Yi Wang , Wei Luo","doi":"10.1016/j.enzmictec.2024.110505","DOIUrl":"10.1016/j.enzmictec.2024.110505","url":null,"abstract":"<div><p>Transaminases (EC 2.6.1.X, TAs) are important biocatalysts in the synthesis of chiral amines, and have significant value in the field of medicine. However, TAs suffer from low enzyme activity and poor catalytic efficiency in the synthesis of chiral amines or non-natural amino acids, which hinders their industrial applications. In this study, a novel TA derived from <em>Paracoccus pantotrophus</em> (<em>pp</em>TA) that was investigated in our previous study was employed with a semi-rational design strategy to improve its enzyme activity to 2-ketobutyrate. By using homology modeling and molecular docking, four surrounding sites in the substrate-binding S pocket were selected as potential mutational sites. Through alanine scanning and saturation mutagenesis, the optimal mutant V153A with significantly improved enzyme activity was finally obtained, which was 578 % higher than that of the wild-type <em>pp</em>TA (WT). Furthermore, the mutant enzyme <em>pp</em>TA-V153A also exhibited slightly improved temperature and pH stability compared to WT. Subsequently, the mutant was used to convert 2-ketobutyrate for the preparation of L-2-aminobutyric acid (L-ABA). The mutant can tolerate 300 mM 2-ketobutyrate with a conversion rate of 74 %, which lays a solid foundation for the preparation of chiral amines.</p></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"180 ","pages":"Article 110505"},"PeriodicalIF":3.4,"publicationDate":"2024-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142083412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Identification of efficient amine transaminase and applicability in dual transaminases cascade for synthesis of L-phosphinothricin 鉴定高效胺转氨酶并将其应用于合成 L-膦丝菌素的双转氨酶级联中

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-08-22 DOI: 10.1016/j.enzmictec.2024.110501

Puhong Yi , Mengdan Liu , Yuhua Hao , Ziwen Wang , Hanlin Liu , Xue Cai , Feng Cheng , Zhiqiang Liu , Yaping Xue , Liqun Jin , Yuguo Zheng

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