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}
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}
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}
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}
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}
{"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}
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}
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}
Boyu Luo , Shanshan Wu , Wei Liu , Dongdong Zhang , Ruicun Liu , Tuoyu Liu , Zhi Sun , Ziqun Wei , Mingyu Liu , Zhiyuan Shi , Niu Huang , Yue Teng
{"title":"Mechanistic insights into the orthogonal functionality of an AHL-mediated quorum-sensing circuit in Yersinia pseudotuberculosis","authors":"Boyu Luo , Shanshan Wu , Wei Liu , Dongdong Zhang , Ruicun Liu , Tuoyu Liu , Zhi Sun , Ziqun Wei , Mingyu Liu , Zhiyuan Shi , Niu Huang , Yue Teng","doi":"10.1016/j.synbio.2024.10.002","DOIUrl":"10.1016/j.synbio.2024.10.002","url":null,"abstract":"<div><div>YpsR, a pivotal regulatory protein in the quorum-sensing (QS) of <em>Yersinia pseudotuberculosis</em>(<em>Y. pstb</em>), is essential for molecular signaling, yet its molecular mechanisms remain poorly understood. Herein, this study systematically investigates the interactions between YpsR and acyl-homoserine lactones (AHLs), shedding light on the selective mechanism of YpsR to various AHL molecules. Using molecular docking and surface plasmon resonance (SPR) analysis, we confirmed YpsR's binding affinities, with the strongest observed for 3OC6-HSL, which notably inhibited <em>Y. pstb</em> growth. Additionally, we engineered a whole-cell biosensor based on YpsR-AHL interaction, which exhibited sensitivity to the signal molecule 3OC6-HSL produced by <em>Y. pstb</em>. Furthermore, key YpsR residues (S32, Y50, W54, D67) involved in AHL binding were identified and validated. Overall, this research elucidates the mechanisms of QS signal recognition in <em>Y. pstb</em>, providing valuable insights that support the development of diagnostic tools for detecting <em>Y. pstb</em> infections.</div></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":"10 1","pages":"Pages 174-184"},"PeriodicalIF":4.4,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142553133","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}