Synthetic and Systems Biotechnology最新文献_第7页

Pathway and protein channel engineering of Bacillus subtilis for improved production of desthiobiotin and biotin 枯草芽孢杆菌改善去硫代生物素和生物素生产的途径和蛋白质通道工程

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-11-22 DOI: 10.1016/j.synbio.2024.11.005

Yue Wu , Guang-Qing Du , Dong-Han Ma , Jin-Long Li , Huan Fang , Hui-Na Dong , Zhao-Xia Jin , Da-Wei Zhang

{"title":"Pathway and protein channel engineering of Bacillus subtilis for improved production of desthiobiotin and biotin","authors":"Yue Wu , Guang-Qing Du , Dong-Han Ma , Jin-Long Li , Huan Fang , Hui-Na Dong , Zhao-Xia Jin , Da-Wei Zhang","doi":"10.1016/j.synbio.2024.11.005","DOIUrl":"10.1016/j.synbio.2024.11.005","url":null,"abstract":"<div><div>Biotin (vitamin B<sub>7</sub>) is a crucial cofactor for various metabolic processes and has significant applications in pharmaceuticals, cosmetics, and animal feed. <em>Bacillus subtilis</em>, a well-studied Gram-positive bacterium, presents a promising host for biotin production due to its Generally Recognized as Safe (GRAS) status, robust genetic tractability, and capacity for metabolite secretion. This study focuses on the metabolic engineering of <em>B</em>. <em>subtilis</em> to enhance biotin biosynthesis. Initially, the desthiobiotin (DTB) and biotin synthesis ability of different <em>B</em>. <em>subtilis</em> strains were evaluated to screen for suitable chassis cells. Subsequently, the titers of DTB and biotin were increased to 21.6 mg/L and 2.7 mg/L, respectively, by relieving the feedback repression of biotin synthesis and deleting the biotin uptake protein YhfU. Finally, through engineering the access tunnel to the active site of biotin synthase (BioB) for reactants and modulating its expression, the biotin titer was increased to 11.2 mg/L, marking an 1130-fold improvement compared to the wild-type strain. These findings provide novel strategies for enhancing the production of DTB and improving the conversion efficiency of DTB to biotin.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 307-313"},"PeriodicalIF":4.4,"publicationDate":"2024-11-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142745797","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

Harnessing microbial heterogeneity for improved biosynthesis fueled by synthetic biology 利用微生物异质性促进合成生物学催化的生物合成

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-11-19 DOI: 10.1016/j.synbio.2024.11.004

Yanting Cao , Jianghua Li , Long Liu , Guocheng Du , Yanfeng Liu

{"title":"Harnessing microbial heterogeneity for improved biosynthesis fueled by synthetic biology","authors":"Yanting Cao , Jianghua Li , Long Liu , Guocheng Du , Yanfeng Liu","doi":"10.1016/j.synbio.2024.11.004","DOIUrl":"10.1016/j.synbio.2024.11.004","url":null,"abstract":"<div><div>Metabolic engineering-driven microbial cell factories have made great progress in the efficient bioproduction of biochemical and recombinant proteins. However, the low efficiency and robustness of microbial cell factories limit their industrial applications. Harnessing microbial heterogeneity contributes to solving this. In this review, the origins of microbial heterogeneity and its effects on biosynthesis are first summarized. Synthetic biology-driven tools and strategies that can be used to improve biosynthesis by increasing and reducing microbial heterogeneity are then systematically summarized. Next, novel single-cell technologies available for unraveling microbial heterogeneity and facilitating heterogeneity regulation are discussed. Furthermore, a combined workflow of increasing genetic heterogeneity in the strain-building step to help in screening highly productive strains - reducing heterogeneity in the production process to obtain highly robust strains (IHP-RHR) facilitated by single-cell technologies was proposed to obtain highly productive and robust strains by harnessing microbial heterogeneity. Finally, the prospects and future challenges are discussed.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 281-293"},"PeriodicalIF":4.4,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142745795","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

Recent advances in the biosynthesis and production optimization of gentamicin: A critical review 庆大霉素生物合成和生产优化的最新进展：重要综述

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-11-14 DOI: 10.1016/j.synbio.2024.11.003

Feng Xu, Kaihao Hu, Ali Mohsin, Jie Wu, Lihuan Su, Yuan Wang, Rong Ben, Hao Gao, Xiwei Tian, Ju Chu

{"title":"Recent advances in the biosynthesis and production optimization of gentamicin: A critical review","authors":"Feng Xu, Kaihao Hu, Ali Mohsin, Jie Wu, Lihuan Su, Yuan Wang, Rong Ben, Hao Gao, Xiwei Tian, Ju Chu","doi":"10.1016/j.synbio.2024.11.003","DOIUrl":"10.1016/j.synbio.2024.11.003","url":null,"abstract":"<div><div>Gentamicin, an aminoglycoside antibiotic, is generated by a few species within the genus <em>Micromonospora</em> and has garnered significant attention due to its broad-spectrum efficacy in combating numerous infectious diseases. Comprising a complex array of closely related aminoglycoside compounds, the gentamicin B and C complexes emerge as particularly pertinent in clinical contexts. This review outlines the latest advancements in the biosynthesis and production of gentamicin, commencing with a comprehensive overview of its biosynthetic pathway. Subsequently, the article encapsulates a spectrum of strategies currently deployed to augment gentamicin yields. These strategies include mutation screening, molecular biological techniques, and optimization of the fermentation process. Moreover, numerous methods have been documented for detecting gentamicin across a range of matrices, underscoring the significance of precise quantitative analysis. Finally, the review furnishes an exhaustive market analysis and future outlook, elucidating prevailing trends and challenges within the gentamicin industry. Overall, this article serves as a pivotal resource for researchers and professionals engaged in gentamicin research, furnishing a meticulous introduction to efficient synthesis technologies and diverse applications, alongside presenting innovative concepts and methodologies aimed at increasing gentamicin production.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 247-261"},"PeriodicalIF":4.4,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142699561","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

Coordinated regulation of two LacI family regulators, GvmR and GvmR2, on guvermectin production in Streptomyces caniferus 两种 LacI 家族调控因子 GvmR 和 GvmR2 对罐头链霉菌生产古维菌素的协调调控

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-11-09 DOI: 10.1016/j.synbio.2024.11.001

Haoran Shi , Jiabin Wang , Shanshan Li , Chongxi Liu , Lei Li , Zhuoxu Dong , Lan Ye , Xiangjing Wang , Yanyan Zhang , Wensheng Xiang

{"title":"Coordinated regulation of two LacI family regulators, GvmR and GvmR2, on guvermectin production in Streptomyces caniferus","authors":"Haoran Shi , Jiabin Wang , Shanshan Li , Chongxi Liu , Lei Li , Zhuoxu Dong , Lan Ye , Xiangjing Wang , Yanyan Zhang , Wensheng Xiang","doi":"10.1016/j.synbio.2024.11.001","DOIUrl":"10.1016/j.synbio.2024.11.001","url":null,"abstract":"<div><div>Guvermectin, a purine nucleoside natural product produced by the genus S<em>treptomyces</em>, has recently been registered as a new biopesticide to boost rice yield. Despite its economic and agricultural significance, the regulatory mechanisms of guvermectin biosynthesis remain essentially unknown, hindering industrial production and widespread agricultural application. Here, we examined the roles of two LacI family regulators, <em>gvmR</em> and <em>gvmR2</em>, located within and adjacent to the guvermectin biosynthesis cluster, respectively, in guvermectin production in <em>Streptomyces caniferus</em> NEAU6. GvmR activated the expression of the guvermectin cluster by binding to the promoters of <em>gvmR</em>, <em>gvmA</em>, and <em>O1</em>, while GvmR2 repressed the guvermectin cluster via competitive binding to promoters containing GvmR-binding sites, specifically, a 14-bp palindromic sequences: 5′-RTCATWCGYATGAY-3′ (R = G/A, W = A/T, Y = T/C). Moreover, GvmR indirectly activates the expression of <em>gvmR2</em> while GvmR2 feedback inhibits <em>gvmR</em> transcription, suggesting a functional interaction between the two regulators for coordinating guvermectin production. Overexpression of <em>gvmR</em> via the T7 RNA polymerase-T7 promoter system in the <em>gvmR2</em> mutant significantly elevated guvermectin production by 125 % (from 631 mg L<sup>−1</sup> to 1422 mg L<sup>−1</sup>), compared to the parental strain NEAU6. This suggested that combinatorial manipulation of <em>gvmR</em> and <em>gvmR2</em> is useful for improving guvermectin production. These findings enrich our knowledge of the regulatory network for guvermectin biosynthesis, and offer key targets and effective strategies for high-titer guvermectin production.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 237-246"},"PeriodicalIF":4.4,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142699560","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

De novo synthesis of 1-phenethylisoquinoline in engineered Escherichia coli 在工程大肠杆菌中从头合成 1-苯乙基异喹啉

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-11-09 DOI: 10.1016/j.synbio.2024.10.007

Yaping Mao , Jiangming Zhu , Qian Zhang , Guangyi Wang , Hongkai Fan , Xiaowei Zhang , Yuwei Sun , Yong Wang

{"title":"De novo synthesis of 1-phenethylisoquinoline in engineered Escherichia coli","authors":"Yaping Mao , Jiangming Zhu , Qian Zhang , Guangyi Wang , Hongkai Fan , Xiaowei Zhang , Yuwei Sun , Yong Wang","doi":"10.1016/j.synbio.2024.10.007","DOIUrl":"10.1016/j.synbio.2024.10.007","url":null,"abstract":"<div><div>Phenylethylisoquinoline alkaloids (PIAs) are medicinally important natural products derived from the 1-phenylethylisoquinoline precursor. Heterologous production of the PIAs remains challenging due to the incomplete elucidation of biosynthetic pathway and the lack of proper microbial cell factory designed for precursor enhancement. In this work, an artificial pathway composed of eight enzymes from different species was established for de novo 1-phenylethylisoquinoline biosynthesis in engineered <em>Escherichia coli</em>. The yield of the intermediate 4-hydroxydihydrocinnamaldehyde was optimized through screening various NADP<sup>+</sup>-dependent 2-alkenal reductases, cofactor regeneration and the site-directed mutagenesis of key residues in ChAER1. Subsequently, incorporation of the modified dopamine pathway into an endogenous reductase-deficient <em>E. coli</em> with high tyrosine yield boosted the production of 1-phenylethylisoquinoline, reaching 402.58 mg/L in a 5L fermenter. Our work lays a foundation for the future large-scale production of high value-added 1-phenylethylisoquinoline-related alkaloids.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 271-280"},"PeriodicalIF":4.4,"publicationDate":"2024-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142699564","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

Harnessing the microbial interactions from Apocynum venetum phyllosphere for natural product discovery 利用芹菜叶球中的微生物相互作用发现天然产品

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-11-08 DOI: 10.1016/j.synbio.2024.11.002

Wei Huang , Xingzhi Jiao , Lingqi Hua , Qianjin Kang , Lili Zhang , Xiaoxia Luo , Linquan Bai

{"title":"Harnessing the microbial interactions from Apocynum venetum phyllosphere for natural product discovery","authors":"Wei Huang , Xingzhi Jiao , Lingqi Hua , Qianjin Kang , Lili Zhang , Xiaoxia Luo , Linquan Bai","doi":"10.1016/j.synbio.2024.11.002","DOIUrl":"10.1016/j.synbio.2024.11.002","url":null,"abstract":"<div><div>Natural products (NPs) afforded by living-beings, especially by microscopic species, represent invaluable and indispensable reservoirs for drug leads in clinical practice. With the rapid advancement in sequencing technology and bioinformatics, the ever-increasing number of microbial biosynthetic gene clusters (BGCs) were decrypted, while a great deal of BGCs remain cryptic or inactive under standard laboratory culture conditions. Addressing this dilemma requires innovative tactics to awaken quiescence of BGCs by releasing the potential of microbial secondary metabolism for mining novel NPs. In this study, a universal strategy was proposed to induce the expression of silent BGCs by leveraging the dynamic interactions among coexisting microbial neighbors within a microbiota. This approach involves the deconstruction/reconstruction of binary interactions among the coexisting neighbors to create a pipeline for BGCs arousing. Coupled with the acquisition of 2760 microbial individuals from the <em>Apocynum venetum</em> (Luobuma, LBM) phyllosphere in a successive dilution procedure, 44 culturable isolates were screened using binary interaction, in which 12.6 % pairs demonstrated potent mutual interacting effects. Furthermore, after selecting the four most promising isolates, a full-scale metabolic inspection was conducted, in which 25.3 % of the interacting pairs showcased significant metabolomic variations with de-cryptic activities. Notably, with the aid of visualization of IMS technology, one of the physiologically functional entities, the bactericidal agent resistomycin, was elucidated from the core interacting pair between the co-culture of the <em>Streptomyces</em> sp. LBM_605 and the <em>Rhodococcus</em> sp. LBM_791. This study highlights the intrinsic interactions among coexisting microorganisms within a phyllosphere microbiota as novel avenues for exploring and harnessing NPs.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 262-270"},"PeriodicalIF":4.4,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142699563","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

Engineering transcriptional regulatory networks for improving second-generation fuel ethanol production in Saccharomyces cerevisiae 改造转录调控网络，提高酿酒酵母的第二代燃料乙醇产量

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-10-28 DOI: 10.1016/j.synbio.2024.10.006

Dongming Sun , Longhao Wu , Xiaocong Lu , Chenhao Li , Lili Xu , Hongxing Li , Deyun He , Aiqun Yu , Tao Yu , Jianzhi Zhao , Hongting Tang , Xiaoming Bao

{"title":"Engineering transcriptional regulatory networks for improving second-generation fuel ethanol production in Saccharomyces cerevisiae","authors":"Dongming Sun , Longhao Wu , Xiaocong Lu , Chenhao Li , Lili Xu , Hongxing Li , Deyun He , Aiqun Yu , Tao Yu , Jianzhi Zhao , Hongting Tang , Xiaoming Bao","doi":"10.1016/j.synbio.2024.10.006","DOIUrl":"10.1016/j.synbio.2024.10.006","url":null,"abstract":"<div><div>Presently, <em>Saccharomyces cerevisiae</em> demonstrates proficient co-fermentation of glucose and xylose, marking a significant advancement in second-generation fuel ethanol production. However, the presence of high concentrations of inhibitors in industrial lignocellulose hydrolysates and post-glucose effect caused by glucose consumption hinders severely impedes yeast robustness and xylose utilization for ethanol fermentation. Even worse, the antagonism between xylose utilization ability and strain robustness was observed, which proposes a difficult challenge in the production of second-generation fuel ethanol by <em>S. cerevisiae</em>. This review introduces the effect of engineering transcriptional regulatory networks on enhancing xylose utilization, improving strain robustness, alleviating antagonism between xylose utilization and strain robustness, and reducing post-glucose effect. Additionally, we provide an outlook on the developmental trends in this field, offering insights into future directions for increasing the production of second-generation fuel ethanol in <em>S. cerevisiae</em>.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 207-217"},"PeriodicalIF":4.4,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142586524","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

Biosensors for the detection of flaviviruses: A review 用于检测黄病毒的生物传感器：综述

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-10-26 DOI: 10.1016/j.synbio.2024.10.005

Ana-Belén Blázquez, Nereida Jiménez de Oya

{"title":"Biosensors for the detection of flaviviruses: A review","authors":"Ana-Belén Blázquez, Nereida Jiménez de Oya","doi":"10.1016/j.synbio.2024.10.005","DOIUrl":"10.1016/j.synbio.2024.10.005","url":null,"abstract":"<div><div>Flaviviruses affect the lives of millions of people in endemic regions and also have the potential to impact non-endemic areas. Factors such as climate change, global warming, deforestation, and increased travel and trade are linked to the spread of flaviviruses into new habitats and host species. Given the absence of specific treatments and the limited availability of vaccines, it is imperative to understand the biology of flaviviruses and develop rapid and sensitive diagnostic tests. These measures are essential for preventing the transmission of these potentially life-threatening pathogens. Flavivirus infections are mainly diagnosed using conventional methods. However, these techniques present several drawbacks, including high expenses, time-consuming procedures, and the need for skilled professionals. The search for fast, easy-to-use, and affordable alternative techniques as a feasible solution for developing countries is leading to the search for new methods in the diagnosis of flaviviruses, such as biosensors.</div><div>This review provides a comprehensive overview of different biosensor detection strategies for flaviviruses and describes recent advances in diagnostic technologies. Finally, we explore their future prospects and potential applications in pathogen detection. This review serves as a valuable resource to understand advances in ongoing research into new biosensor-based diagnostic methods for flaviviruses.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 194-206"},"PeriodicalIF":4.4,"publicationDate":"2024-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142561287","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

Systematic metabolic engineering of Yarrowia lipolytica for efficient production of phytohormone abscisic acid 利用脂肪分解酵母的系统代谢工程高效生产植物激素脱落酸

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-10-18 DOI: 10.1016/j.synbio.2024.10.004

Mei-Li Sun , Ziyun Zou , Lu Lin , Rodrigo Ledesma-Amaro , Kaifeng Wang , Xiao-Jun Ji

{"title":"Systematic metabolic engineering of Yarrowia lipolytica for efficient production of phytohormone abscisic acid","authors":"Mei-Li Sun , Ziyun Zou , Lu Lin , Rodrigo Ledesma-Amaro , Kaifeng Wang , Xiao-Jun Ji","doi":"10.1016/j.synbio.2024.10.004","DOIUrl":"10.1016/j.synbio.2024.10.004","url":null,"abstract":"<div><div>Abscisic acid (ABA) is an important phytohormone with diverse applications. It currently relies on the fermentation of <em>Botrytis cinerea</em>, which suffers from limited availability of genetic engineering tools. Here, <em>Yarrowia lipolytica</em> was engineered to enable <em>de novo</em> biosynthesis of ABA. To overcome the rate-limiting P450 enzymes, systematic engineering strategies were implemented. Firstly, the dissolved oxygen was increased to boost the activity of P450 enzymes. Secondly, the expansion of endoplasmic reticulum was implemented to improve the functional expression of P450 enzymes. Lastly, rate-limiting enzymes were assembled to facilitate substrate trafficking. Moreover, ABA production was further improved by strengthening the mevalonate pathway. Finally, the engineered strain produced 1221.45 mg/L of ABA in a 5-L bioreactor. The study provides effective approaches for alleviating rate-limiting P450 enzymes to enhance ABA production and achieve competitive industrial-level ABA production in <em>Y. lipolytica</em>.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 165-173"},"PeriodicalIF":4.4,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142553132","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

Enhancing substrate specificity of microbial transglutaminase for precise nanobody labeling 增强微生物转谷氨酰胺酶的底物特异性，实现精确的纳米抗体标记

IF 4.4 2区生物学

Synthetic and Systems Biotechnology Pub Date : 2024-10-16 DOI: 10.1016/j.synbio.2024.10.003

Xinglong Wang , Kangjie Xu , Haoran Fu , Qiming Chen , Beichen Zhao , Xinyi Zhao , Jingwen Zhou

{"title":"Enhancing substrate specificity of microbial transglutaminase for precise nanobody labeling","authors":"Xinglong Wang , Kangjie Xu , Haoran Fu , Qiming Chen , Beichen Zhao , Xinyi Zhao , Jingwen Zhou","doi":"10.1016/j.synbio.2024.10.003","DOIUrl":"10.1016/j.synbio.2024.10.003","url":null,"abstract":"<div><div><em>Streptomyces mobaraenesis</em> transglutaminase (smTG) can be used for site-specific labeling of proteins with chemical groups. Here, we explored the use of modified smTG for the biosynthesis of nanobody-fluorophore conjugates (NFC). smTG catalyzes the conjugation of acyl donors containing glutamine with lysine-containing acceptors, which can lead to non-specific cross-linking. To achieve precise site-specific labeling, we employed molecular docking and virtual mutagenesis to redesign the enzyme's substrate specificity towards the peptide GGGGQR, a non-preferred acyl donor for smTG. Starting with a thermostable and highly active smTG variant (TGm2), we identified that single mutations G250H and Y278E significantly enhanced activity against GGGGQR, increasing it by 41 % and 1.13-fold, respectively. Notably, the Y278E mutation dramatically shifted the enzyme's substrate preference, with the activity ratio against GGGGQR versus the standard substrate CBZ-Gln-Gly rising from 0.05 to 0.93. In case studies, we used nanobodies 1C12 and 7D12 as labeling targets, catalyzing their conjugation with a synthetic fluorophore via smTG variants. Nanobodies fused with GGGGQR were successfully site-specifically labeled by TGm2-Y278E, in contrast to non-specific labeling observed with other variants. These results suggest that engineering smTG for site-specific labeling is a promising approach for the biosynthesis of antibody-drug conjugates.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 185-193"},"PeriodicalIF":4.4,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142553134","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