Journal of biotechnology最新文献_第9页

Fecal microbiota transplantation combined with inulin promotes the development and function of early immune organs in chicks 粪菌群移植联合菊粉可促进雏鸡早期免疫器官的发育和功能。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-16 DOI: 10.1016/j.jbiotec.2025.01.012

Yang Song, Yibo Cui, Yue Zhong, Yumeng Wang, Xin Zheng

{"title":"Fecal microbiota transplantation combined with inulin promotes the development and function of early immune organs in chicks","authors":"Yang Song, Yibo Cui, Yue Zhong, Yumeng Wang, Xin Zheng","doi":"10.1016/j.jbiotec.2025.01.012","DOIUrl":"10.1016/j.jbiotec.2025.01.012","url":null,"abstract":"<div><div>Modern management of chicks hinders the vertical transmission of intestinal microbiota, which is closely related to immunity. Inulin is a substrate that can be utilized by the microbiota. This study aimed to determine whether fecal microbiota transplantation (FMT) combined with inulin played a \"1 + 1 > 2\" role in enhancing the development and function of immune organs. Chicks were treated with 1 % inulin and/or fecal microbiota suspension on days 1–6. The growth performance, immune organ development, and immune indicators were evaluated on days 7, 14, and 21. Results showed that the combination of FMT and inulin significantly increased the immune organ index on day 7 and promoted the morphological structure and the expression of proliferating cell nuclear antigen (PCNA) in immune organs on days 7, 14, and 21. Each treatment increased the gene expression of interferon-gamma (IFN-γ), interleukin-4 (IL-4), interleukin-2 (IL-2), B cell-activating factor receptor (BAFFR), B cell linker (BLNK), C-X-C Motif Chemokine Ligand 12 (CXCL12), C-X-C Motif Chemokine Receptor 4 (CXCR4), and Biotin (Bu-1) to varying degrees. FMT combined with inulin significantly increased the expression of IgA-positive cells on days 7 and 14. In conclusion, the synergistic effect of FMT and inulin had beneficial impacts on the development and function of immune organs.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 81-90"},"PeriodicalIF":4.1,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005981","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Investigating subpopulation dynamics in clonal CHO-K1 cells with single-cell RNA sequencing 单细胞RNA测序研究克隆CHO-K1细胞亚群动态。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-15 DOI: 10.1016/j.jbiotec.2025.01.010

Luke B. Morina , Haoyu Chris Cao , Siqi Chen , Swetha Kumar , Kevin S. McFarland , Natalia I. Majewska , Michael J. Betenbaugh , Winston Timp

{"title":"Investigating subpopulation dynamics in clonal CHO-K1 cells with single-cell RNA sequencing","authors":"Luke B. Morina , Haoyu Chris Cao , Siqi Chen , Swetha Kumar , Kevin S. McFarland , Natalia I. Majewska , Michael J. Betenbaugh , Winston Timp","doi":"10.1016/j.jbiotec.2025.01.010","DOIUrl":"10.1016/j.jbiotec.2025.01.010","url":null,"abstract":"<div><div>Chinese Hamster Ovary (CHO) cells produce monoclonal antibodies and other biotherapeutics at industrial scale. Despite their ubiquitous nature in the biopharmaceutical industry, little is known about the behaviors of individual transfected clonal CHO cells. Most CHO cells are assessed on their stability, their ability to produce the protein of interest over time. But CHO cells have primarily been studied in bulk, instead assuming that these bulk samples are homogenous because of presumed genetic clonality across the sample. This does not address cellular heterogeneity in these ostensibly clonal cells. These variable stability phenotypes may reflect heterogeneity within the clonal samples. In this study, we performed single-cell RNA sequencing on two clonal CHO-K1 cell populations with different stability phenotypes over a 90 day culture period. Our data showed that the instability of one of the clone’s gene expression was due in part to the emergence of a low-producing subpopulation in the aged samples. This low-producing subpopulation did not exhibit markers of cellular stress which were expressed in the higher-producing populations. Further multiomic investigation should be performed to better characterize this heterogeneity.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 91-98"},"PeriodicalIF":4.1,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Interferon inhibitors increase rAAV production in HEK293 cells 干扰素抑制剂增加HEK293细胞中rAAV的产生。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-15 DOI: 10.1016/j.jbiotec.2025.01.009

Yongdan Wang , Qiang Fu , Sha Sha , Seongkyu Yoon

{"title":"Interferon inhibitors increase rAAV production in HEK293 cells","authors":"Yongdan Wang , Qiang Fu , Sha Sha , Seongkyu Yoon","doi":"10.1016/j.jbiotec.2025.01.009","DOIUrl":"10.1016/j.jbiotec.2025.01.009","url":null,"abstract":"<div><div>Recombinant adeno-associated viruses (rAAVs) comprise a promising viral vector for therapeutic gene delivery to treat disease. However, the current manufacturing capability of rAAVs must be improved to meet commercial demand. Previously published omics studies indicate that rAAV production through transient transfection triggers antiviral responses and endoplasmic reticulum stress responses in the host cell. Both responses negatively regulate viral production. We demonstrate that the modulation of the antiviral immune response (by blocking interferon signaling pathways) can effectively lower the production of interferon and enhance viral genome production. The use of interferon inhibitors before transfection can significantly increase rAAV production in HEK293 cells, with up to a 2-fold increase in productivity and up to a 6-fold increase in specific productivity. Compared to the untreated groups, the addition of these small molecules generally reduced viable cell density but increased vector productivity. The positive candidates were BX795 (a TBK inhibitor), TPCA-1 (an IKK2 inhibitor), Cyt387 (a JAK1 inhibitor), and ruxolitinib (another JAK1 inhibitor). These candidates were identified using deep well screening, and reproducible titer improvement was achieved in a 30 mL shake flask scale. Additionally, genome titer improvement is feasible and scalable in two different media, but the extent of improvement may vary.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 9-18"},"PeriodicalIF":4.1,"publicationDate":"2025-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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 highly efficient mixed strain fermentation strategy to produce 11α-Hydroxyandrost-4-ene-3,17-dione from phytosterols 植物甾醇高效混合发酵生产11α-羟基雄激素-4-烯-3,17-二酮的研究。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-14 DOI: 10.1016/j.jbiotec.2025.01.007

Zhenhua Su, Yanfei Li, Chang Shi, Dantong Liu, Yan Yang, Yanbing Shen, Min Wang

{"title":"A highly efficient mixed strain fermentation strategy to produce 11α-Hydroxyandrost-4-ene-3,17-dione from phytosterols","authors":"Zhenhua Su, Yanfei Li, Chang Shi, Dantong Liu, Yan Yang, Yanbing Shen, Min Wang","doi":"10.1016/j.jbiotec.2025.01.007","DOIUrl":"10.1016/j.jbiotec.2025.01.007","url":null,"abstract":"<div><div>11α-Hydroxyandrost-4-ene-3,17-dione (11α-OH AD) is an essential steroid hormone drug intermediate that exhibits low biotransformation efficiency. In this study, a mixed-strain fermentation strategy was established for the efficient production of 11α-OH AD from phytosterols (PS). Initially, strains were screened for efficient transformation of AD to produce 11α-OH AD. Subsequently, a dual-strain mixed-culture fermentation technique was established, with <em>Mycolicibacterium neoaurum</em> CICC 21097 ΔksdD (MNR) showing highly effective results. Ultimately, a one-step conversion process for the production of 11α-OH AD was achieved at a molar yield of 76.5 % under optimal conditions using PS as a substrate, the highest reported yield to date. Additionally, studies revealed synergistic metabolic interactions between MNR and <em>Aspergillus ochraceus</em> in the mixed-culture system. These findings provide valuable insights for the industrial production of high-value products using mixed-strain fermentation.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 1-8"},"PeriodicalIF":4.1,"publicationDate":"2025-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Structure-guided mining of stereoselective reductive aminases for biocatalytic stereodivergent synthesis of chiral piperidinamine and derivatives 手性哌啶胺及其衍生物生物催化立体发散合成中立体选择性还原氨基酶的结构导向挖掘。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-13 DOI: 10.1016/j.jbiotec.2025.01.004

Wen Zhang , Xiangyu Zheng , Yudong Hu , Ye Ni , Guochao Xu

{"title":"Structure-guided mining of stereoselective reductive aminases for biocatalytic stereodivergent synthesis of chiral piperidinamine and derivatives","authors":"Wen Zhang , Xiangyu Zheng , Yudong Hu , Ye Ni , Guochao Xu","doi":"10.1016/j.jbiotec.2025.01.004","DOIUrl":"10.1016/j.jbiotec.2025.01.004","url":null,"abstract":"<div><div>Chiral azacyclic amine derivatives occupy a vital role of nitrogen-containing compounds, due to serve as foundational motifs in numerous pharmaceuticals and bioactive substances. Novel complementary enantioselective reductive aminases IRED9 and IRED11 were unveiled through comprehensive gene mining from <em>Streptomyces viridochromogenes</em> and <em>Micromonospora echinaurantiaca</em>, respectively, which both demonstrated enantiomeric excess (<em>ee</em>) values and conversion ratios of up to 99 % towards <em>N</em>-Boc-3-pyridinone (NBPO) and cyclopropylamine. IRED9 exhibited the highest activity at pH 8.0 and 45 °C，while IRED11 have optimal conditions at pH 8.0 and 50 °C. A variety of amine donors and ketones could be converted by IRED9 and IRED11 for asymmetric synthesis of piperidinamine and derivatives with complementary enantioselectivity. Through preparative-scale synthesis of (<em>S</em>)- and (<em>R</em>)-3-piperidinamine, IRED9 and IRED11 demonstrate substrate loadings of 120 g·L<sup>–1</sup> and 40 g·L<sup>–1</sup> with 98 % yield and 99 % <em>ee</em>, respectively. The space time yield (STY) reached 142.7 g·L<sup>–1</sup>d<sup>–1</sup> and 47.1 g·L<sup>–1</sup>d<sup>–1</sup> for the <em>S</em> enantiomer and <em>R</em> enantiomer, respectively. Interaction analysis indicated the substrate orientation and strong charge attraction interaction are vital factors for enantioselectivity of IREDs. This study unveils novel enantioselective reductive aminases for stereodivergent synthesis of piperidinamine and derivatives at high substrate loading.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 28-37"},"PeriodicalIF":4.1,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005993","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Ni2+-induced selective precipitation of His-tagged recombinant proteins shortens purification time while maintaining high yield Ni2+诱导的his标记重组蛋白选择性沉淀在保持高产的同时缩短了纯化时间。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-13 DOI: 10.1016/j.jbiotec.2025.01.006

Md. Din Islam , M. Monirul Islam , Yutaka Kuroda

{"title":"Ni2+-induced selective precipitation of His-tagged recombinant proteins shortens purification time while maintaining high yield","authors":"Md. Din Islam , M. Monirul Islam , Yutaka Kuroda","doi":"10.1016/j.jbiotec.2025.01.006","DOIUrl":"10.1016/j.jbiotec.2025.01.006","url":null,"abstract":"<div><div>Nickel-NTA affinity chromatography is the current standard method for purifying His-tagged recombinant proteins. However, this process involves repetitive tasks, can be time-consuming, and reduces protein yield. Here, we present a simple, fast, and handy method for purifying His-tagged proteins using free Ni²⁺. This approach allows the fractional precipitation of His-tagged proteins directly from <em>E. coli</em> cell lysates. We successfully applied this Ni²⁺-based method to purify three His₆-tagged recombinant proteins overexpressed in <em>E. coli</em>. We found that Ni²⁺ at a final concentration of as low as 1 mM precipitates the His-tagged proteins with near-complete specificity as confirmed by SDS-PAGE analysis. The Ni²<sup>+</sup>-precipitated proteins were dissolved by adding 10 % acetic acid and further purified by reverse-phase HPLC. The final yields were between 3.5 and 8.0 mg per 200 mL culture, similar to or even higher than purification using conventional Ni-NTA chromatography. The purified proteins exhibited natively folded characteristics, as assessed by CD, SLS, and DLS, and binding activity, as assessed by ELISA and BLI, demonstrating the method's potential in both small and large-scale settings.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 38-46"},"PeriodicalIF":4.1,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Engineering silica nanocoated whole-cell asymmetric biocatalyst for efficient preparation of a key chiral intermediate of (S)-Rivastigmine 工程二氧化硅纳米包被全细胞不对称生物催化剂高效制备(S)-利瓦斯汀关键手性中间体。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-13 DOI: 10.1016/j.jbiotec.2025.01.005

Baoling Chen, Hang Yang, Ruixuan Bai, Xiaotong Du, Yue Gao, Liangyu Zheng

{"title":"Engineering silica nanocoated whole-cell asymmetric biocatalyst for efficient preparation of a key chiral intermediate of (S)-Rivastigmine","authors":"Baoling Chen, Hang Yang, Ruixuan Bai, Xiaotong Du, Yue Gao, Liangyu Zheng","doi":"10.1016/j.jbiotec.2025.01.005","DOIUrl":"10.1016/j.jbiotec.2025.01.005","url":null,"abstract":"<div><div>In our previous study, the whole cells containing an aldo–keto reductase (yhdN) and glucose dehydrogenase (GDH) were constructed and applied in a stereoselective carbonyl reduction reaction to prepare (<em>S</em>)-NEMCA-HEPE, being a key chiral intermediate of (<em>S</em>)-Rivastigmine which is widely prescribed for the treatment of Alzheimer’s disease. Although the conversion and enantiomeric excess (<em>e.e.</em>) could reach to 78.2 % and 99 %, respectively, ionic liquid as an additive was required to improve the permeability of cell membrane. To further simplify the reaction, the molecular docking and saturation mutagenesis technology were used here to obtain an activity-improved yhdN variant such as G19A. And then, both excellent conversion and <em>e.e.</em> of 99 % for (<em>S</em>)-NEMCA-HEPE could be achieved within 40 min by using only G19A-GDH whole cell as a catalyst without any additive. However, the use of the whole cells still faces the issues of poor operation stability and adverse application prospect. Subsequently, a hydrophobic \"cell-in-shell\" complex of G19A-GDH@O-Silica was constructed by using a silica nanocoated technology. The obtained G19A-GDH@O-Silica exhibited an excellent conversion towards the asymmetric carbonyl reduction, and a good tolerance in changing thermal, pH, and storage environmental. Giving 76.3 % of reaction conversion even after the 11th cycle of reuse, indicated that G19A-GDH@O-Silica also possessed ideal recyclability. The aim of this study is to provide a rapid, and cost-effective nanocoated whole-cell biocatalyst for efficient preparation of (<em>S</em>)-NEMCA-HEPE. The simplicity and robustness of the immobilization approach may become a powerful tool to utilize whole-cell catalysts towards organic catalysis.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 19-27"},"PeriodicalIF":4.1,"publicationDate":"2025-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143005990","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Immobilization of glycosyltransferase into a hydrophilic metal-organic framework for efficient biosynthesis of chondroitin sulfate 糖基转移酶在亲水金属-有机框架中的固定化用于硫酸软骨素的高效生物合成。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2025-01-10 DOI: 10.1016/j.jbiotec.2025.01.003

Xinyue Zhang , Yanqi Li , Jingjing Bi , Junjie Zhang , Bingzhi Li , Xing Zhang , Jie Zheng , Lei Lin

{"title":"Immobilization of glycosyltransferase into a hydrophilic metal-organic framework for efficient biosynthesis of chondroitin sulfate","authors":"Xinyue Zhang , Yanqi Li , Jingjing Bi , Junjie Zhang , Bingzhi Li , Xing Zhang , Jie Zheng , Lei Lin","doi":"10.1016/j.jbiotec.2025.01.003","DOIUrl":"10.1016/j.jbiotec.2025.01.003","url":null,"abstract":"<div><div>Chondroitin sulfate (CS) is a structurally complex anionic polysaccharide widely used in medical, cosmetic and food applications. Enzymatic catalysis is an important strategy for synthesizing CS with uniform chain lengths and well-defined structures. However, the industrial application of glycosyltransferases is hindered by limitations such as low expression yields, poor stability, and challenges in reuse. We developed a mild and rapid one-step synthetic method for the efficient immobilization of chondroitin synthase (KfoC). The resulting KfoC@ZIF-90 composite exhibits high catalytic activity, thermal stability, and pH adaptability. Notably, KfoC@ZIF-90 exhibited 5-fold enhanced thermal stability at 40°C and retained 86 % relative activity at pH 10, while also maintaining 90 % activity in organic solvents, surpassing the performance of free KfoC. Molecular docking analysis revealed that the binding capability of encapsulated KfoC with substrate was stronger than that of free KfoC, thereby improving catalytic performance. Furthermore, KfoC@ZIF-90 can be easily separated from the reaction solution by centrifugation, simplifying product isolation and purification while enabling enzyme reuse. These attributes significantly enhance operability and reduce processing costs, making enzymatic CS synthesis more feasible for industrial applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"399 ","pages":"Pages 63-71"},"PeriodicalIF":4.1,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142970698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Assessing the role of deep eutectic solvents in Yarrowia lipolytica inhibition 评估深共晶溶剂在抑制多脂耶氏菌中的作用

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.016

Filipe S. Buarque , Bernardo D. Ribeiro , Mara G. Freire , Maria A.Z. Coelho , Matheus M. Pereira

{"title":"Assessing the role of deep eutectic solvents in Yarrowia lipolytica inhibition","authors":"Filipe S. Buarque , Bernardo D. Ribeiro , Mara G. Freire , Maria A.Z. Coelho , Matheus M. Pereira","doi":"10.1016/j.jbiotec.2024.11.016","DOIUrl":"10.1016/j.jbiotec.2024.11.016","url":null,"abstract":"<div><div><em>Yarrowia lipolytica</em> has gained recognition as a microorganism with biological relevance and extensive biotechnological applications. Some of its features include a high enzyme secretion capacity and a high cell-density fermentation mode. Hexokinase (YlHxk) is a vital enzyme in <em>Y. lipolytica</em> growth since it catalyzes glucose metabolism through phosphorylation in the glycolytic pathway. Given the potential application of deep eutectic solvents (DES) as novel solvents in biotechnological processes, this study evaluated the influence of eighteen DES on the growth of <em>Y. lipolytica</em>. Furthermore, this work examined the effects of individual ions on the YlHxk enzyme by analyzing its enzymatic tunnel structure, molecule transport, and molecular docking. The results revealed a significant reduction in yeast growth in the presence of most DES compared to the control (medium without DES), with the exception of the [N<sub>8881</sub>]Cl: hexanoic acid (1:1) DES. The growth varied between 11.95 ± 0.60 and 0.68 ± 0.17 g dry cell weight L<sup>−1</sup>. According to the enzymatic tunnel analysis, DES components associated with the lowest microbial growth values were transported through tunnel 1. On the other hand, DES components had their pathway facilitated through tunnel 2 ([N<sub>8881</sub>]<sup>+</sup> and hexanoic acid) and showed growth values close to the control. Molecular docking analysis identified a similarity between all the ligands in this tunnel (including substrate and product), presenting binding interactions with the ASN273 amino acid of the YlHxk active site. Combining experimental results with computational tools provided promising insights at the molecular level, while also potentially reducing analysis costs and time, paving the way for similar approaches in broad biocatalytic reactions.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"398 ","pages":"Pages 1-10"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142748017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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

Creation of catalytic activity-improved hyperthermophilic PQQ-dependent aldose sugar dehydrogenase and its efficient use for high performance electro-device 催化活性改进的超耐热pqq依赖性醛糖脱氢酶的建立及其在高性能电器件中的高效应用。

IF 4.1 2区生物学

Journal of biotechnology Pub Date : 2024-11-28 DOI: 10.1016/j.jbiotec.2024.11.017

Miku Maeno , Yusuke Miki , Kazuki Ito , Haruhiko Sakuraba , Toshihisa Ohshima , Shin-ichiro Suye , Takenori Satomura

{"title":"Creation of catalytic activity-improved hyperthermophilic PQQ-dependent aldose sugar dehydrogenase and its efficient use for high performance electro-device","authors":"Miku Maeno , Yusuke Miki , Kazuki Ito , Haruhiko Sakuraba , Toshihisa Ohshima , Shin-ichiro Suye , Takenori Satomura","doi":"10.1016/j.jbiotec.2024.11.017","DOIUrl":"10.1016/j.jbiotec.2024.11.017","url":null,"abstract":"<div><div>PQQ-dependent aldose sugar dehydrogenase (PQQ-ASD) from the hyperthermophilic archaeon <em>Pyrobaculum aerophilum</em> (PaeASD) has great potential as an element for durable bioelectrodevices owing to its exceptional stability against high temperatures and across a broad pH spectrum. However, its application is constrained by low electric current output of the enzyme-immobilized electrodes, which is attributable to its low catalytic activity. A directed evolutionary approach was performed on PaeASD to improve enzyme activity, resulting in the identification of a PaeASD s24 mutant containing six amino acid substitutions, which exhibited a 16-fold higher specific activity than that of wild type. Although each single amino acid mutant among these substitutions exhibited lower enzyme activity than PaeASD s24, the double mutant R64Q/D350N showed enzyme activity comparable to that of PaeASD s24. These amino acids located in the vicinity of coenzyme PQQ within the PaeASD molecule are also highly conserved with those of PQQ-ASDs reported to date. Thus, these amino acids play crucial roles in the catalytic activity of PQQ-ASD. Furthermore, the <em>K</em><sub>m</sub> value for <span>d</span>-glucose of PaeASD s24-immobilized electrode decreased to approximately 1/3 that of the wild-type-immobilized electrode. These results indicate that the PaeASD s24 mutant is an excellent catalyst for potential bioelectrodevice applications.</div></div>","PeriodicalId":15153,"journal":{"name":"Journal of biotechnology","volume":"398 ","pages":"Pages 11-17"},"PeriodicalIF":4.1,"publicationDate":"2024-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142755169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"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