Enzyme and Microbial Technology最新文献_第9页

Facile benzothiazole-triazole based thiazole derivatives as novel thymidine phosphorylase and α-glucosidase inhibitors: Experimental and computational approaches 以苯并噻唑-三唑为基础的噻唑衍生物作为新型胸苷磷酸化酶和α-葡萄糖苷酶抑制剂的简便方法：实验和计算方法

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-06-13 DOI: 10.1016/j.enzmictec.2024.110470

Shoaib Khan , Rafaqat Hussain , Yousaf Khan , Tayyiaba Iqbal , Farman Ullah , Shifa Felemban , M.M. Khowdiary

{"title":"Facile benzothiazole-triazole based thiazole derivatives as novel thymidine phosphorylase and α-glucosidase inhibitors: Experimental and computational approaches","authors":"Shoaib Khan , Rafaqat Hussain , Yousaf Khan , Tayyiaba Iqbal , Farman Ullah , Shifa Felemban , M.M. Khowdiary","doi":"10.1016/j.enzmictec.2024.110470","DOIUrl":"10.1016/j.enzmictec.2024.110470","url":null,"abstract":"<div>The present study reports the new thiazole (A-L) derivatives based on benzothiazole fused triazole which were synthesized and assessed against thymidine phosphorylase and α-glucosidase enzymes. Several compounds with the same basic structure but different substituents were found to have high activity against the targeted enzymes, while others with the same basic skeleton but different substituents were found to have medium to low activity among the members of tested series. These analogs showed a varied range of inhibition in both case thymidine phosphorylase and alpha glucosidase, A (IC50 = 7.20 ± 0.30 µM and IC50 = 1.30 ± 0.70 µM), B (IC50 = 8.80 ± 0.10 µM and IC50 = 2.10 ± 0.30 µM), C (IC50 = 8.90 ± 0.40 µM and IC50 = 3.20 ± 0.20 µM) and thiazole containing analogs such as G (IC50 = 11.10 ± 0.20 µM and IC50 = 7.80 ± 0.20 µM) and H (IC50 = 12.30 ± 0.30 µM and IC50 = 6.30 ± 0.20 µM). When compared with standard drugs 7-Deazaxanthine, 7DX (IC50 = 10.60 ± 0.50 µM) and acarbose (IC50 = 4.30 ± 0.30 µM) respectively. These analogs were also subjected to molecular docking studies which indicated the binding interaction of molecules with active sites of the enzyme and strengthen the drug profile of these compounds. ADMET studies also predict the drug-like properties of these compounds, with no violations of drug likeness rules.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110470"},"PeriodicalIF":3.4,"publicationDate":"2024-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141391413","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

Whole-cell biocatalysis for ε-poly-l-lysine production by a food-grade recombinant Bacillus subtilis 利用食品级重组枯草芽孢杆菌的全细胞生物催化技术生产ε-聚赖氨酸

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-06-07 DOI: 10.1016/j.enzmictec.2024.110467

Kunpeng Li , Yangzi Guo , Xinjie Sun , Xiangheng Xi , Li Wang , Xidong Ren , Chenying Wang , Xinli Liu

{"title":"Whole-cell biocatalysis for ε-poly-l-lysine production by a food-grade recombinant Bacillus subtilis","authors":"Kunpeng Li , Yangzi Guo , Xinjie Sun , Xiangheng Xi , Li Wang , Xidong Ren , Chenying Wang , Xinli Liu","doi":"10.1016/j.enzmictec.2024.110467","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110467","url":null,"abstract":"<div>ε-Poly-l-lysine (ε-PL), a natural food preservative with various advantages, is primarily produced by Streptomyces. It has attracted considerable attentions for the outstanding antibacterial activity, safety, heat stability, water solubility and other remarkable properties. In this study, a food-grade recombinant Bacillus subtilis was constructed for the biocatalysis of ε-PL. Firstly, the d-alanine racemase gene (alrA) was deleted from the genome of Bacillus subtilis 168 to construct an auxotrophic B. subtilis 168 (alrA-). Based on the shuttle plasmid pMA5, a food-grade plasmid pMA5a was constructed by replacing the genes of kanamycin resistance (Kanr) and ampicillin resistance (Ampr) with alrA and the gene encoding α-peptide of β-galactosidase (lacZα), respectively. Subsequently, codon-optimized ε-PL synthase gene (pls) and P-pls were ligated into pMA5a and transformed in E. coli DH5α and expressed in B. subtilis 168 (alrA-). Finally, the whole-cell biocatalysis conditions for ε-PL production by B. subtilis 168 (alrA-)/pMA5a-pls were optimized, and the optimal conditions were 30°C, pH 4, l-lysine concentration of 0.6 g/L, bacterial concentration of 15 % (w/v) and a catalytic time of 7 h. The ε-PL production reached a maximum of 0.33 ± 0.03 g/L. The product was verified to be ε-PL by HPLC and tricine-SDS-PAGE. The information obtained in this study shows critical reference for the food-grade heterologous expression of ε-PL.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110467"},"PeriodicalIF":3.4,"publicationDate":"2024-06-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141289834","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

Development of whole cell biocatalytic system for asymmetric synthesis of esomeprazole with enhancing coenzyme biosynthesis pathway 利用增强型辅酶生物合成途径开发不对称合成埃索美拉唑的全细胞生物催化系统

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-06-06 DOI: 10.1016/j.enzmictec.2024.110469

Xinqi Xu, Yaping Meng, Bingmei Su, Juan Lin

{"title":"Development of whole cell biocatalytic system for asymmetric synthesis of esomeprazole with enhancing coenzyme biosynthesis pathway","authors":"Xinqi Xu, Yaping Meng, Bingmei Su, Juan Lin","doi":"10.1016/j.enzmictec.2024.110469","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110469","url":null,"abstract":"<div>Esomeprazole is the most popular proton pump inhibitor for treating gastroesophageal reflux disease. Previously, a phenylacetone monooxygenase mutant LnPAMOmu15 (LM15) was obtained by protein engineering for asymmetric synthesis of esomeprazole using pyrmetazole as substrate. To scale up the whole cell asymmetric synthesis of esomeprazole and reduce the cost, in this work, an Escherichia coli whole-cell catalyst harboring LM15 and formate dehydrogenase from Burkholderia stabilis 15516 (BstFDH) were constructed through optimized gene assembly patterns. CRISPR/Cas9 mediated insertion of Ptrc promoter in genome was done to enhance the expression of key genes to increase the cellular NADP supply in the whole cell catalyst, by which the amount of externally added NADP+ for the asymmetric synthesis of esomeprazole decreased to 0.05 mM from 0.3 mM for reducing the cost. After the optimization of reaction conditions in the reactor, the scalable synthesis of esomeprazole was performed using the efficient LM15-BstFDH whole-cell as catalyst, which showed the highest reported space-time yield of 3.28 g/L/h with 50 mM of pyrmetazole loading. Isolation procedure was conducted to obtain esomeprazole sodium of 99.55 % purity and > 99.9 % ee with 90.1 % isolation yield. This work provides the basis for production of enantio-pure esomeprazole via cost-effective whole cell biocatalysis.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110469"},"PeriodicalIF":3.4,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141323189","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

Insights on kraft lignin degradation in an anaerobic environment 在厌氧环境中降解牛皮纸木质素的启示

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-06-03 DOI: 10.1016/j.enzmictec.2024.110468

Jéssica P. Silva , Tayná D. Frederico , Alonso R.P. Ticona , Otávio H.B. Pinto , Thomas C.R. Williams , Ricardo H. Krüger , Eliane F. Noronha

{"title":"Insights on kraft lignin degradation in an anaerobic environment","authors":"Jéssica P. Silva , Tayná D. Frederico , Alonso R.P. Ticona , Otávio H.B. Pinto , Thomas C.R. Williams , Ricardo H. Krüger , Eliane F. Noronha","doi":"10.1016/j.enzmictec.2024.110468","DOIUrl":"10.1016/j.enzmictec.2024.110468","url":null,"abstract":"<div>Lignin is an aromatic macromolecule and one of the main constituents of lignocellulosic materials. Kraft lignin is generated as a residual by-product of the lignocellulosic biomass industrial process, and it might be used as a feedstock to generate low molecular weight aromatic compounds. In this study, we seek to understand and explore the potential of ruminal bacteria in the degradation of kraft lignin. We established two consortia, KLY and KL, which demonstrated significant lignin-degrading capabilities. Both consortia reached maximum growth after two days, with KLY showing a higher growth and decolorization rate. Additionally, SEM analysis revealed morphological changes in the residual lignin from both consortia, indicating significant degradation. This was further supported by FTIR spectra, which showed new bands corresponding to the C-H vibrations of guaiacyl and syringyl units, suggesting structural transformations of the lignin. Taxonomic analysis showed enrichment of the microbial community with members of the Dickeya genus. Seven metabolic pathways related to lignin metabolism were predicted for the established consortia. Both consortia were capable of consuming aromatic compounds such as 4-hydroxybenzoic acid, syringaldehyde, acetovanillone, and syringic acid, highlighting their capacity to convert aromatic compounds into commercially valuable molecules presenting antifungal activity and used as food preservatives as 4-hydroxyphenylacetic, 3-phenylacetic, and phenylacetic acids. Therefore, the microbial consortia shown in the present work are models for understanding the process of lignin degradation and consumption in bacterial anaerobic communities and developing biological processes to add value to industrial processes based on lignocellulosic biomass as feedstock.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110468"},"PeriodicalIF":3.4,"publicationDate":"2024-06-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141275949","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

Biochemical identification of D-mannose 2-epimerase from Cytophagaceae bacterium SJW1-29 for efficient bioconversion of D-glucose to D-mannose 从噬菌体细菌 SJW1-29 中鉴定 D-甘露糖 2-酰亚胺酶的生化特性，以实现 D-葡萄糖到 D-甘露糖的高效生物转化

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-06-02 DOI: 10.1016/j.enzmictec.2024.110465

Dawei Ni , Yuhan Wei , Yulei Zhang , Tarek A.A. Moussa , Wenli Zhang , Wanmeng Mu

{"title":"Biochemical identification of D-mannose 2-epimerase from Cytophagaceae bacterium SJW1-29 for efficient bioconversion of D-glucose to D-mannose","authors":"Dawei Ni , Yuhan Wei , Yulei Zhang , Tarek A.A. Moussa , Wenli Zhang , Wanmeng Mu","doi":"10.1016/j.enzmictec.2024.110465","DOIUrl":"10.1016/j.enzmictec.2024.110465","url":null,"abstract":"<div>Enzymatic production of D-mannose attracts increasing attention because of the health effects and commercial values of D-mannose. Several kinds of epimerases or isomerases have been used for enzymatic production of D-mannose from D-glucose or D-fructose. D-Mannose epimerase (MEase), belonging to N-acyl-D-glucosamine 2-epimerase superfamily enzymes, catalyzes the C-2 epimerization between D-glucose and D-mannose. In this study, a novel MEase was identified from Cytophagaceae bacterium SJW1-29. Sequence and structure alignments indicate that it is highly conserved with the reported R. slithyformis MEase with the known crystal structure. It was a metal-independent enzyme, with an optimal pH of 8.0 and an optimal temperature of 40 °C. The specific activities on D-glucose and D-mannose were 2.90 and 2.96 U/mg, respectively. The Km, kcat, and kcat/Km on D-glucose were measured to be 194.9 mM, 2.72 s−1, and 0.014 mM−1 s−1, respectively. The purified enzyme produced 23.15 g/L of D-mannose from 100 g/L of D-glucose at pH 8.0 and 40 °C for 8 h, with a conversion rate of 23.15 %.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110465"},"PeriodicalIF":3.4,"publicationDate":"2024-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141276664","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

Development of a chloroplast expression system for Dunaliella salina 为杜纳利藻开发叶绿体表达系统

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-05-27 DOI: 10.1016/j.enzmictec.2024.110464

Hao-Hong Chen , Qian-Xi Zheng , Fan Yu , Shan-Rong Xie , Jian-Guo Jiang

{"title":"Development of a chloroplast expression system for Dunaliella salina","authors":"Hao-Hong Chen , Qian-Xi Zheng , Fan Yu , Shan-Rong Xie , Jian-Guo Jiang","doi":"10.1016/j.enzmictec.2024.110464","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110464","url":null,"abstract":"<div>Dunaliella salina is an innovative expression system due to its distinct advantages such as high salt tolerance, low susceptibility to contamination, and the absence of the cell wall. While nuclear transformation has been extensively studied, research on D. salina chloroplast transformation remains in the preliminary stages. In this study, we established an efficient chloroplast expression system for D. salina using Golden Gate assembly. We developed a D. salina toolkit comprising essential components such as chloroplast-specific promoters, terminators, homologous fragments, and various vectors. We confirmed its functionality by expressing the EGFP protein. Moreover, we detailed the methodology of the entire construction process. This expression system enables the specific targeting of foreign genes through simple homologous recombination, resulting in stable expression in chloroplasts. The toolkit achieved a relatively high transformation efficiency within a shorter experimental cycle. Consequently, the construction and utilization of this toolkit have the potential to enhance the efficiency of transgenic engineering in D. salina and advance the development of microalgal biofactories.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110464"},"PeriodicalIF":3.4,"publicationDate":"2024-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141286463","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

Antioxidant capacity of xylooligosaccharides generated from beechwood xylan by recombinant family GH10 Aspergillus niger xylanase A and insights into the enzyme's competitive inhibition by riceXIP 重组 GH10 家族黑曲霉木聚糖酶 A 从榉木木聚糖生成的木寡糖的抗氧化能力以及对该酶受水稻 XIP 竞争性抑制的深入研究

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-05-12 DOI: 10.1016/j.enzmictec.2024.110456

Keer Zhang, Xinyu Qi, Ningxin Feng, Yuzhu Wang, Huiwen Wei, Mingqi Liu

{"title":"Antioxidant capacity of xylooligosaccharides generated from beechwood xylan by recombinant family GH10 Aspergillus niger xylanase A and insights into the enzyme's competitive inhibition by riceXIP","authors":"Keer Zhang, Xinyu Qi, Ningxin Feng, Yuzhu Wang, Huiwen Wei, Mingqi Liu","doi":"10.1016/j.enzmictec.2024.110456","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110456","url":null,"abstract":"<div>In this study, the family GH10 xylanase AnXylA10 derived from Aspergillus niger JL15 strain was expressed in Pichia pastoris X33. The recombinant xylanase, reAnXylA10 exhibited optimal activity at 40 ℃ and pH 5.0. The hydrolysates generated from beechwood xylan using reAnXylA10 primarily consisted of xylobiose (X2) to xylohexaose (X6) and demonstrated remarkable antioxidant capacity. Furthermore, the rice xylanase inhibitory protein (riceXIP) was observed to competitively inhibit reAnXylA10, exhibiting an inhibition constant (Ki) of 140.6 nM. Molecular dynamics (MD) simulations of AnXylA10-riceXIP complex revealed that the α-7 helix (Q225-S238) of riceXIP intruded into the catalytic pocket of AnXylA10, thereby obstructing substrate access to the active site. Specifically, residue K226 of riceXIP formed robust interactions with E136 and E242, the two catalytic sites of AnXylA10, predominantly through high-occupied hydrogen bonds. Based on QTAIM, electron densities for the atom pairs K226riceXIP@HZ1-E136AnXylA10@OE2 and K226riceXIP@HZ3-E242AnXylA10@OE1 were determined to be 0.04628 and 0.02914 a.u., respectively. Binding free energy of AnXylA10-riceXIP complex was −59.0±7.6 kcal/mol, significantly driven by electrostatic and van der Waals forces. Gaining insights into the interaction between xylanase and its inhibitors, and mining the inhibition mechanism in depth, will facilitate the design of innovative GH10 family xylanases that are both highly efficient and resistant to inhibitors.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"179 ","pages":"Article 110456"},"PeriodicalIF":3.4,"publicationDate":"2024-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140947311","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

Whole-cell bioconversion for producing thymoquinone by engineered Saccharomyces cerevisiae 利用工程酿酒酵母进行全细胞生物转化生产胸腺醌

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-04-26 DOI: 10.1016/j.enzmictec.2024.110455

Eunjee Kim, Minsun Kim, Min-Kyu Oh

{"title":"Whole-cell bioconversion for producing thymoquinone by engineered Saccharomyces cerevisiae","authors":"Eunjee Kim, Minsun Kim, Min-Kyu Oh","doi":"10.1016/j.enzmictec.2024.110455","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110455","url":null,"abstract":"<div>Thymoquinone, extracted from the black seeds of Nigella sativa, is a natural substance with highly beneficial effects against various human diseases. In this study, we aimed to construct a Saccharomyces cerevisiae strain that, produce thymoquinone from thymol, a relatively inexpensive substrate. To achieve this, cytochrome P450 from Origanum vulgare was expressed in S. cerevisiae for the bioconversion of thymol to thymoquinone, with the co-expression of cytochrome P450 reductase (CPR) from Arabidopsis thaliana, ATR1. Additionally, flexible linkers were used to connect these two enzymes. Furthermore, modifications were performed to expand the endoplasmic reticulum (ER) space, leading to increased thymoquinone production. After integrating the genes into the chromosome and optimizing the media components, a significant improvement in the thymol-to-thymoquinone conversion rate and yield were achieved. This study represents a possibility of the production of thymoquinone, a bioactive ingredient of a plant, using an engineered microbial cell.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"178 ","pages":"Article 110455"},"PeriodicalIF":3.4,"publicationDate":"2024-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140893919","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

Rational design improves both thermostability and activity of a new D-tagatose 3-epimerase from Kroppenstedtia eburnean to produce D-allulose 合理设计提高了 Kroppenstedtia eburnean 中一种新的 D-tagatose 3-epimerase 的热稳定性和活性，以生产 D-allulose

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-04-17 DOI: 10.1016/j.enzmictec.2024.110448

Dingyu Guo , Zhengchao Wang , Wanqing Wei , Wei Song , Jing Wu , Jian Wen , Guipeng Hu , Xiaomin Li , Cong Gao , Xiulai Chen , Liming Liu

{"title":"Rational design improves both thermostability and activity of a new D-tagatose 3-epimerase from Kroppenstedtia eburnean to produce D-allulose","authors":"Dingyu Guo , Zhengchao Wang , Wanqing Wei , Wei Song , Jing Wu , Jian Wen , Guipeng Hu , Xiaomin Li , Cong Gao , Xiulai Chen , Liming Liu","doi":"10.1016/j.enzmictec.2024.110448","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110448","url":null,"abstract":"<div>D-allulose is a naturally occurring rare sugar and beneficial to human health. However, the efficient biosynthesis of D-allulose remains a challenge. Here, we mined a new D-tagatose 3-epimerase from Kroppenstedtia eburnean (KeDt3e) with high catalytic efficiency. Initially, crucial factors contributing to the low conversion of KeDt3e were identified through crystal structure analysis, density functional theory calculations (DFT), and molecular dynamics (MD) simulations. Subsequently, based on the mechanism, combining restructuring the flexible region, proline substitution based onconsensus sequence analysis, introducing disulfide bonds, and grafting properties, and reshaping the active center, the optimal mutant M5 of KeDt3e was obtained with enhanced thermostability and activity. The optimal mutant M5 exhibited an enzyme activity of 130.8 U/mg, representing a 1.2-fold increase; Tm value increased from 52.7 °C to 71.2 °C; and half-life at 55 °C extended to 273.7 min, representing a 58.2-fold improvement, and the detailed mechanism of performance improvement was analyzed. Finally, by screening the ribosome-binding site (RBS) of the optimal mutant M5 recombinant bacterium (G01), the engineered strain G08 with higher expression levels was obtained. The engineered strain G08 catalyzed 500 g/L D-fructose to produce 172.4 g/L D-allulose, with a conversion of 34.4% in 0.5 h and productivity of 344.8 g/L/h on a 1 L scale. This study presents a promising approach for industrial-scale production of D-allulose.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"178 ","pages":"Article 110448"},"PeriodicalIF":3.4,"publicationDate":"2024-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140641040","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

Physiological and biochemical characteristics of the carbon ion beam irradiation-generated mutant strain Clostridium butyricum FZM 240 in vitro and in vivo 碳离子束辐照产生的突变菌株丁酸梭菌 FZM 240 在体外和体内的生理生化特征

IF 3.4 3区生物学

Enzyme and Microbial Technology Pub Date : 2024-04-15 DOI: 10.1016/j.enzmictec.2024.110447

Ya-Juan Wang , Xiang Zhou , Miao-Miao Zhang , Mei-Han Liu , Nan Ding , Qing-Feng Wu , Cai-Rong Lei , Zi-Yi Dong , Jun-Le Ren , Jing-Ru Zhao , Cheng-Lin Jia , Jun Liu , Bo Zhou , Dong Lu

{"title":"Physiological and biochemical characteristics of the carbon ion beam irradiation-generated mutant strain Clostridium butyricum FZM 240 in vitro and in vivo","authors":"Ya-Juan Wang , Xiang Zhou , Miao-Miao Zhang , Mei-Han Liu , Nan Ding , Qing-Feng Wu , Cai-Rong Lei , Zi-Yi Dong , Jun-Le Ren , Jing-Ru Zhao , Cheng-Lin Jia , Jun Liu , Bo Zhou , Dong Lu","doi":"10.1016/j.enzmictec.2024.110447","DOIUrl":"https://doi.org/10.1016/j.enzmictec.2024.110447","url":null,"abstract":"<div>Clostridium butyricum (C. butyricum) represents a new generation of probiotics, which is beneficial because of its good tolerance and ability to produce beneficial metabolites, such as short-chain fatty acids and enzymes; however, its low enzyme activity limits its probiotic efficacy. In this study, a mutant strain, C. butyricum FZM 240 was obtained using carbon ion beam irradiation, which exhibited greatly improved enzyme production and tolerance. The highest filter paper, endoglucanase, and amylase activities produced by C. butyricum FZM 240 were 125.69 U/mL, 225.82 U/ mL, and 252.28 U/mL, which were 2.58, 1.95, and 2.21-fold higher, respectively, than those of the original strain. The survival rate of the strain increased by 11.40 % and 5.60 % after incubation at 90 °C for 5 min and with simulated gastric fluid at pH 2.5 for 2 h, respectively, compared with that of the original strain. Whole-genome resequencing and quantitative real-time PCR(qRT-PCR) analysis showed that the expression of genes related to enzyme synthesis (GE000348, GE001963 and GE003123) and tolerance (GE001114) was significantly up-regulated, while that of genes related to acid metabolism (GE003450) was significantly down-regulated. On this basis, homology modeling and functional prediction of the proteins encoded by the mutated genes were performed. According to the results, the properties related to the efficacy of C. butyricum as a probiotic were significantly enhanced by carbon ion beam irradiation, which is a novel strategy for the application of Clostridium spp. as feed additives.</div>","PeriodicalId":11770,"journal":{"name":"Enzyme and Microbial Technology","volume":"178 ","pages":"Article 110447"},"PeriodicalIF":3.4,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140555621","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}

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