Enzyme and Microbial Technology最新文献

Effect of lipopolysaccharide on ganglioside expression in human induced pluripotent stem cell-derived kidney organoids. 脂多糖对人诱导多能干细胞衍生肾类器官中神经节苷脂表达的影响。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-12-01 Epub Date: 2025-08-06 DOI: 10.1016/j.enzmictec.2025.110730

Jae-Sung Ryu, Jin Ok Yu, Ki Kwan Kim, Eun-Jeong Jeong, Min Young Kim, Hyo Gyeong Yun, Eun Bin Song, Ji-Su Kim, Young-Woock Noh, Young-Kug Choo

{"title":"Effect of lipopolysaccharide on ganglioside expression in human induced pluripotent stem cell-derived kidney organoids.","authors":"Jae-Sung Ryu, Jin Ok Yu, Ki Kwan Kim, Eun-Jeong Jeong, Min Young Kim, Hyo Gyeong Yun, Eun Bin Song, Ji-Su Kim, Young-Woock Noh, Young-Kug Choo","doi":"10.1016/j.enzmictec.2025.110730","DOIUrl":"10.1016/j.enzmictec.2025.110730","url":null,"abstract":"<p><p>An organoid is a self-organizing, three-dimensional (3D), stem cell-derived structure that closely mimics the structural, cellular, and functional properties of specific organs or tissues. Organoids are widely utilized for assessing drug efficacy, safety, and industrial chemical toxicity. The purpose of this study was to generate a kidney organoid from human induced pluripotent stem cells (iPSCs) and establish a sepsis-associated acute kidney injury (SA-AKI) model by treatment with lipopolysaccharide (LPS). We further analyzed changes in ganglioside expression following LPS treatment in kidney organoids. As a result, we observed that the expression of kidney-specific markers was significantly increased during differentiation. Next, we confirmed that the levels of inflammation-related markers and reactive oxygen species (ROS) were significantly increased, whereas mitochondrial membrane potential (MMPΨ) was significantly reduced in LPS-treated kidney organoids. Interestingly, ganglioside GM3, GM2, GD3, and GD1a expression, as well as their biosynthesis, was notably decreased in LPS-treated kidney organoids. These findings suggest that gangliosides play critical roles in inflammation and may contribute to the pathophysiology of SA-AKI, highlighting the potential of kidney organoids as a valuable model system for studying kidney injury and associated inflammatory responses.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"110730"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144816100","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

Modulating dioxygenase and hydroperoxide isomerase activities in Burkholderia thailandensis lipoxygenase. 调节泰国伯克霍尔德菌脂加氧酶的双加氧酶和氢过氧化物异构酶活性。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-12-01 Epub Date: 2025-07-08 DOI: 10.1016/j.enzmictec.2025.110709

Ruth Chrisnasari, Roelant Hilgers, Guanna Li, Jean-Paul Vincken, Willem J H van Berkel, Marie Hennebelle, Tom A Ewing

{"title":"Modulating dioxygenase and hydroperoxide isomerase activities in Burkholderia thailandensis lipoxygenase.","authors":"Ruth Chrisnasari, Roelant Hilgers, Guanna Li, Jean-Paul Vincken, Willem J H van Berkel, Marie Hennebelle, Tom A Ewing","doi":"10.1016/j.enzmictec.2025.110709","DOIUrl":"10.1016/j.enzmictec.2025.110709","url":null,"abstract":"<p><p>Lipoxygenases (LOXs) are enzymes that catalyze the regioselective dioxygenation of polyunsaturated fatty acids (PUFAs), leading to the formation of fatty acid hydroperoxides (FAHPs). In addition to dioxygenase activity, some eukaryotic LOXs exhibit hydroperoxide isomerase (HPI) activity under specific conditions, resulting in the production of structurally diverse compounds such as epoxy alcohols and ketones. Until now, the presence of HPI activity in bacterial LOXs has not been documented. In this study, we investigated the HPI activity of LOX from Burkholderia thailandensis (Bt-LOX) and examined the effects of reaction conditions on its catalytic profile using three different C18 PUFA substrates. The results demonstrated that Bt-LOX exhibits significant HPI activity, especially at high enzyme concentrations, with ketone formation showing strong substrate dependence. Oxygen level was identified as a critical factor in directing the catalytic performance of Bt-LOX: HPI activity was inhibited under O₂-saturated conditions and enhanced under O₂-limited conditions. These findings establish Bt-LOX as the first bacterial LOX reported to exhibit pronounced HPI activity, and highlights its expanded potential for biocatalytic applications.</p>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"191 ","pages":"110709"},"PeriodicalIF":3.7,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144811958","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

Mechanism of tyrosine 69 in the flavin domain modulating electron transfer efficiency in BVU5 azoreductase 黄素区域酪氨酸69调节BVU5偶氮还原酶电子转移效率的机制

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-25 DOI: 10.1016/j.enzmictec.2025.110757

Xuehui Xie , Dongyang Li , Hangmi Zheng , Ziyi Wu , Jiajie Liu , Wanting Li , Songwei Chen , Yiwen Zhang , Daijie Tang , Na Liu , Qingyun Zhang , Ye Chen

{"title":"Mechanism of tyrosine 69 in the flavin domain modulating electron transfer efficiency in BVU5 azoreductase","authors":"Xuehui Xie , Dongyang Li , Hangmi Zheng , Ziyi Wu , Jiajie Liu , Wanting Li , Songwei Chen , Yiwen Zhang , Daijie Tang , Na Liu , Qingyun Zhang , Ye Chen","doi":"10.1016/j.enzmictec.2025.110757","DOIUrl":"10.1016/j.enzmictec.2025.110757","url":null,"abstract":"<div><div>The azoreductase BVU5 is a flavoprotein dependent on NAD(P)H/FMN-mediated electron transfer; However, the functional mechanism of key residues within its flavin domain remains unclear. This study identified the conserved residue Tyr<sup>69</sup> through bioinformatics analysis and constructed Y69F and Y69C mutants. Enzymatic assays demonstrated that mutants exhibited significantly lower decolorization rates than the BVU5 enzyme across 12 dye molecules, including azo, anthraquinone, and triphenylmethane dyes. For example, when decolorizing Reactive Black 5 (RB5) for 2 h: Y69F mutant achieved 60 %-65 % decolorization, Y69C mutant reached 55 %-60 % decolorization, both markedly lower than the 80 %-85 % efficiency of BVU5.The decolorization hierarchy remained azo dyes > triphenylmethane > anthraquinone dyes. Molecular docking revealed that mutations reconfigured FMN-binding patterns. Although Y69F enlarged the substrate-binding pocket, it failed to enhance the degradation efficiency of the bulky dye Chlorazol Black E. This critical contradiction indicates that substrate binding is not the limiting factor. Combined with evidence such as the lighter color of the mutant enzyme solutions, increased A280/A450 ratios, and enzyme dosage experiments, this study confirms that Tyr<sup>69</sup> is a key residue that sustains electron transfer efficiency by maintaining the FMN-binding conformation, thereby determining the decolorization performance. Consequently, electron transfer efficiency, rather than substrate binding, is the primary mechanism influencing the catalytic function of BVU5.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110757"},"PeriodicalIF":3.7,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145155168","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 a thermostable GH6 family cellulase from chaetomium thermophilum exhibiting high cellobiose and ionic liquid tolerance 嗜热毛菌GH6家族耐热纤维素酶的鉴定，具有高纤维二糖和离子液体耐受性

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-23 DOI: 10.1016/j.enzmictec.2025.110755

Pooja , Sushant K. Sinha , Supratim Datta

{"title":"Identification of a thermostable GH6 family cellulase from chaetomium thermophilum exhibiting high cellobiose and ionic liquid tolerance","authors":"Pooja , Sushant K. Sinha , Supratim Datta","doi":"10.1016/j.enzmictec.2025.110755","DOIUrl":"10.1016/j.enzmictec.2025.110755","url":null,"abstract":"<div><div>Efficient conversion of lignocellulosic biomass into fermentable sugars requires cellulases that are both thermostable and tolerant to inhibitors such as ionic liquids (ILs). Thermostable enzymes are particularly valuable for industrial applications, as they maintain activity at elevated temperatures for extended periods, improve product yield, and reduce process costs. In this study, we cloned and characterized <em>Ct</em>Cel6C, a GH6 family cellulase, from the thermophilic fungus <em>Thermochaetoides thermophila</em> (<em>Chaetomium thermophilum</em>). <em>Ct</em>Cel6C exhibited a specific activity of 28 U/mg on carboxymethyl cellulose (CMC-Na) at 55 °C and pH 6.0, retaining 90 % of its specific activity after 20 h at optimum pH and temperature (55 °C and pH 6.0). <em>Ct</em>Cel6C demonstrated broad substrate specificity, effectively hydrolyzing oligosaccharides, soluble substrates (CMC and barley β-glucan), and insoluble substrates such as Avicel, consistently producing cellobiose as the sole product and outperforming other GH6 cellobiohydrolases from <em>C. thermophilum</em>. Despite having an open active-site cleft due to a lack of a fifteen-residue stretch in the C-terminal loop, <em>Ct</em>Cel6C retains the biochemical characteristics of a processive cellobiohydrolase rather than those of an endoglucanase from the GH6 family. The enzyme also exhibited high tolerance to cellobiose, glucose, and to 20 % (v/v) of the ionic liquid 1-ethyl-3-methylimidazolium acetate ([C<sub>2</sub>mim][OAc]), in both McIlvaine buffer (pH 6.0) and seawater. The remarkable IL tolerance of <em>Ct</em>Cel6C makes it a promising candidate for integration into industrial enzyme cocktails for biomass saccharification.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110755"},"PeriodicalIF":3.7,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145217523","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

Preparation of magnetic xylanase cross-linked aggregates (m-CLXAs) for the hydrolysis of arabinoxylan by mechano-enzymology 机械酶水解阿拉伯木聚糖的磁性木聚糖酶交联聚集体（m-CLXAs）制备。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-23 DOI: 10.1016/j.enzmictec.2025.110756

Justin B. Safari , Lebogang Ramatsui , Liam Nisbet , Vincent J. Smith , Rui W.M. Krause , Brett I. Pletschke

{"title":"Preparation of magnetic xylanase cross-linked aggregates (m-CLXAs) for the hydrolysis of arabinoxylan by mechano-enzymology","authors":"Justin B. Safari , Lebogang Ramatsui , Liam Nisbet , Vincent J. Smith , Rui W.M. Krause , Brett I. Pletschke","doi":"10.1016/j.enzmictec.2025.110756","DOIUrl":"10.1016/j.enzmictec.2025.110756","url":null,"abstract":"<div><div>The development of a biocatalyst consisting of immobilised xylanases as cross-linked enzyme aggregates (CLEAs) on magnetic nanoparticles (MNPs) as support was undertaken. MNPs were prepared by a coprecipitation reaction of Fe<sup>3 +</sup> and Fe<sup>2+</sup>, followed by surface modification with (3-aminopropyl)-trimethoxysilane using a polycondensation reaction. Magnetic CLEAs were prepared via glutaraldehyde cross-linking after precipitation with acetone, and the physicochemical characterisation of the immobilised enzyme was performed at all stages using several techniques (FTIR, PXRD, TGA, and DLS). A one-factor-at-a-time approach (OFAT) was used to investigate the impact of temperature, time, enzyme: MNP ratio, precipitant, and cross-linking agent to determine their effect on the enzyme’s recovered activity. The results demonstrated that all parameters impacted the immobilisation differently, with the optimised conditions determined as 4°C, 12 h, 10 mg/g, 60 % (v/v) acetone, and 200 mM glutaraldehyde, respectively. The immobilisation did not affect the pH and temperature optima of the enzyme, which were 6.0 and 50–70°C, respectively, for both the free and immobilised enzymes. In contrast, the immobilised enzyme could be reused more than ten times to hydrolyse wheat arabinoxylan, without losing 50 % of its initial activity. Values for <em>V</em><sub><em>max</em></sub> and <em>K</em><sub><em>m</em></sub> only decreased slightly compared to those obtained for the free enzyme. The thermal inactivation parameters showed that the immobilisation procedure did not adversely affect the enzyme's catalytic properties after immobilisation. Finally, we assessed the immobilised enzyme for its ability to catalyse reactions under mechanochemical conditions (grinding and ageing) and found that the free and immobilised enzymes were active during solvent-free and liquid-assisted grinding (LAG).</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110756"},"PeriodicalIF":3.7,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145198823","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

Enhanced synthesis of alanyl-glutamine dipeptide via molecular modification and process optimization of α-amino acid ester acyltransferase EAET α-氨基酸酯酰基转移酶EAET的分子修饰和工艺优化促进丙氨酰-谷氨酰胺二肽的合成。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-22 DOI: 10.1016/j.enzmictec.2025.110754

Jinao Wei , Xiaoxi Liu , Yifei Gao, Jing-wen Yang, Hong-bin Zhang, Xue-qin Hu

{"title":"Enhanced synthesis of alanyl-glutamine dipeptide via molecular modification and process optimization of α-amino acid ester acyltransferase EAET","authors":"Jinao Wei , Xiaoxi Liu , Yifei Gao, Jing-wen Yang, Hong-bin Zhang, Xue-qin Hu","doi":"10.1016/j.enzmictec.2025.110754","DOIUrl":"10.1016/j.enzmictec.2025.110754","url":null,"abstract":"<div><div>This study presents a significant advancement in the enzymatic synthesis of alanyl-glutamine (Ala-Gln) through the development of a high-performance α-ester acyltransferase (EAET) using molecular engineering. We successfully cloned EAET from <em>Bacillus pumilus</em>, which exhibits low sequence identity (<30 %) compared to other reported acyltransferases (AETs). Using AlphaFold3, we constructed a highly reliable structural model of EAET, validated by metrics such as pLDDT, ipTM, and PAE. Through semi-rational design, we introduced the F330Y mutation, which enhanced enzymatic activity by 1.5-fold and extended the half-life at 30°C by 1.3-fold. Molecular dynamics simulations revealed increased flexibility around residue 330 and additional hydrogen bonds in the F330Y mutant, both of which contribute to its improved catalytic efficiency and thermostability. Response surface methodology was employed to optimize enzyme production conditions, resulting in a maximum enzyme activity of 218.20 U/mL[91.3 U/(mL·OD600)]. Systematic optimization of catalytic parameters, including pH, temperature, substrate ratio, concentration, enzyme dosage, and reaction time, enabled an 82.59 % conversion yield at 300 mM substrate concentration. These results not only surpass previous benchmarks but also demonstrate the industrial potential of engineered EAET variants for Ala-Gln synthesis. This study highlights the successful integration of computational tools with molecular engineering to enhance enzyme performance, offering a greener and more sustainable approach for Ala-Gln production.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110754"},"PeriodicalIF":3.7,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145205964","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

A thermostable cellobiose phosphorylase from Thermoclostridium caenicola and its application to in vitro biotransformation 一种耐热性纤维二糖磷酸化酶及其在体外生物转化中的应用

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-19 DOI: 10.1016/j.enzmictec.2025.110753

Haotian Cheng , Yuan Li , Yi Zhou , Shuke Wu , Yi-Heng P. Job Zhang

{"title":"A thermostable cellobiose phosphorylase from Thermoclostridium caenicola and its application to in vitro biotransformation","authors":"Haotian Cheng , Yuan Li , Yi Zhou , Shuke Wu , Yi-Heng P. Job Zhang","doi":"10.1016/j.enzmictec.2025.110753","DOIUrl":"10.1016/j.enzmictec.2025.110753","url":null,"abstract":"<div><div>Cellobiose phosphorylase (CBP) catalyzes the reversible phosphorolysis of cellobiose to glucose and α-D-glucose-1-phosphate. CBPs play an important role in the <em>in vivo</em> enzymatic utilization of cellulosic materials. Here we discovered a novel CBP from <em>Thermoclostridium caenicola</em> (TcCBP), displaying 50–75 % sequence homology with previously reported CBPs. Recombinant TcCBP was well expressed in <em>E. coli</em> BL21(DE3), with a 1.6-fold increase in soluble expression levels over the widely-used <em>Clostridium thermocellum</em> CBP (CtCBP). This enzyme exhibited broad pH adaptability, maintaining substantial activities across pH 4.0–7.5 in the synthetic direction and pH 5.0–7.5 in the phosphorolytic direction. Compared to CtCBP, TcCBP displayed the superior thermostability and a nearly 100-fold improvement in \"Product-to-Enzyme Ratio\" (PE value). The biosynthesis of <em>myo</em>-inositol from cellobiose was conducted by using this CBP along with other four thermophilic enzymes (i.e., phosphoglucomutase, inositol 1-phosphate synthase, inositol monophosphatase, and polyphosphate glucokinase) in one pot without step-by-step addition of enzymes. Approximately 96 mM of <em>myo</em>-inositol was produced from 50 mM of cellobiose. These results indicated that this enzyme could be a potential thermophilic enzyme used for the production of value-added biochemicals by <em>in vitro</em> BioTransformation.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110753"},"PeriodicalIF":3.7,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145109831","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

Strong alone, weak together: biofilm tensile strength in kangaroo rat burrows 单独强，一起弱：袋鼠大鼠洞中的生物膜拉伸强度

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-17 DOI: 10.1016/j.enzmictec.2025.110752

Duygu Aydin , Idil Deniz Akin , Douglas R. Call , Haluk Beyenal

{"title":"Strong alone, weak together: biofilm tensile strength in kangaroo rat burrows","authors":"Duygu Aydin , Idil Deniz Akin , Douglas R. Call , Haluk Beyenal","doi":"10.1016/j.enzmictec.2025.110752","DOIUrl":"10.1016/j.enzmictec.2025.110752","url":null,"abstract":"<div><div>Desert kangaroo rats (<em>Dipodomys deserti</em>) construct burrows that create protected micro-niches favorable to increased microbial activity and biofilm formation. Biofilms within these burrows bind sand particles together, increase the tensile strength of the burrow sand and burrow stability. Our previous work 1) demonstrated that kangaroo rat burrow sand exhibits higher tensile strength than surrounding surface sand due to the cementation by biofilms, and 2) characterized the microbial biofilm communities within kangaroo rat burrow sand and isolated abundant microorganisms. In this study, isolated species are used to quantify tensile strength of pure and mixed biofilms using the microcantilever technique. Mono-species biofilms of <em>Aspergillus tamarii</em> (59.30 ± 4.36 kPa) exhibited the highest tensile strength, while <em>Neobacillus niacini</em> (9.45 ± 1.98 kPa) showed the lowest. Dual-species biofilms displayed synergistic or antagonistic effects, depending on species combinations. Mixing <em>N. niacini</em> with <em>Peribacillus frigoritolerans</em> increased tensile strength to 55.11 ± 2.51 kPa, whereas combining <em>A. tamarii</em> with <em>N. niacini</em> reduced the tensile strength to 18.98 ± 2.54 kPa. Mixing up to five species reduced tensile strength to a minimum value of 2.16 kPa. We conclude that biofilms formed by microbial isolates from burrow sand individually had higher tensile strength, but when all were mixed, the tensile strength decreased, making them weaker.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110752"},"PeriodicalIF":3.7,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145106315","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

De Novo enzyme design of a minimal and soluble carbonic anhydrase from Mesorhizobium loti assisted by molecular dynamics simulation 在分子动力学模拟的辅助下，从loti中分离出一种最小的可溶碳酸酐酶

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-12 DOI: 10.1016/j.enzmictec.2025.110751

Jiun-Jang Juo, I.-Son Ng

{"title":"De Novo enzyme design of a minimal and soluble carbonic anhydrase from Mesorhizobium loti assisted by molecular dynamics simulation","authors":"Jiun-Jang Juo, I.-Son Ng","doi":"10.1016/j.enzmictec.2025.110751","DOIUrl":"10.1016/j.enzmictec.2025.110751","url":null,"abstract":"<div><div>Carbonic anhydrases (CAs) are critical biocatalysts in the carbon capture and utilization due to their remarkable efficiency in converting carbon dioxide into bicarbonate. Among all, the CA from <em>Mesorhizobium loti</em> (MlCA) exhibits the highest catalytic activity. However, the poor expression and low solubility in <em>Escherichia coli</em> significantly restricts its application. To overcome the challenges, we employed a protein minimization strategy to improve both enzymatic expression and solubility. Enhanced expressions were observed in genetic constructs with short N-terminal tags which MlCA expression is strongly affected by mRNA secondary structure near the start codon. For solubility issue, a <em>de novo</em> protein design workflow guided by molecular dynamics simulations was developed. The process consists of four stages: (1) size-constrained <em>de novo</em> design considered using AlphaFold and RFdiffusion, (2) sequence recovery replying on solubleMPNN and ESMfold, (3) <em>In silico</em> screening by FoldX, SASA and molecular dynamics evaluation, and (4) experimental validation. This enabled the generation of compact, stable, and catalytically active MlCA variants with 28 % reduction in protein size among 500 candidates. Finally, the most promising design, dM22, of minimal CA showed improved solubility from 16.1 % to 61.2 % in B7G, a GroELS integrated BL21(DE3) strain.</div></div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"192 ","pages":"Article 110751"},"PeriodicalIF":3.7,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145057156","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

Corrigendum to "Influence of a rumen ciliate-derived xylanase on the gut microbiota composition: A potential enzyme for prebiotic applications" [Enzyme Microb. Technol. 190 (2025) 110683]. “瘤胃纤毛虫衍生的木聚糖酶对肠道微生物群组成的影响：一种潜在的益生元应用酶”的更正[微生物酶]。技术通报，190 (2025)110683 [j]。

IF 3.7 3区生物学

Enzyme and Microbial Technology Pub Date : 2025-09-10 DOI: 10.1016/j.enzmictec.2025.110749

Weide Su, Huiying Luo, Gaoxiang Ai, Qipeng Wei, Zhiheng Zou, Xiaolian Chen, Chuanhui Xu, Jiang Chen, Pingwen Xiong, Wenjing Song, Qiongli Song

引用次数: 0