Synthetic and Systems Biotechnology最新文献

{"title":"Minimizing endogenous cryptic plasmids to construct antibiotic-free expression systems for Escherichia coli Nissle 1917","authors":"Siyan Zhou ,&nbsp;Linlin Zhao ,&nbsp;Wenjie Zuo ,&nbsp;Yilin Zheng ,&nbsp;Ping Zhang ,&nbsp;Yanan Sun ,&nbsp;Yang Wang ,&nbsp;Guocheng Du ,&nbsp;Zhen Kang","doi":"10.1016/j.synbio.2024.01.006","DOIUrl":"10.1016/j.synbio.2024.01.006","url":null,"abstract":"<div><p>The probiotic bacterium <em>Escherichia coli</em> Nissle 1917 (EcN) holds significant promise for use in clinical and biological industries. However, the reliance on antibiotics to maintain plasmid-borne genes has overshadowed its benefits. In this study, we addressed this issue by engineering the endogenous cryptic plasmids pMUT1 and pMUT2. The non-essential elements were removed to create more stable derivatives pMUT1NR△ and pMUT2HBC△. Synthetic promoters by integrating binding motifs on sigma factors were further constructed and applied for expression of <em>Bacteroides thetaiotaomicron</em> heparinase III and the biosynthesis of ectoine. Compared to traditional antibiotic-dependent expression systems, our newly constructed antibiotic-free expression systems offer considerable advantages for clinical and synthetic biology applications.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000061/pdfft?md5=b44bddea0817611a5f60c572baea48a2&pid=1-s2.0-S2405805X24000061-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139635250","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":"Nitric oxide synthase-guided genome mining identifies a cytochrome P450 enzyme for olefin nitration in bacterial specialized metabolism","authors":"Hu Li ,&nbsp;Wei Li ,&nbsp;Kaihui Song ,&nbsp;Yu Liu ,&nbsp;Guiyun Zhao ,&nbsp;Yi-Ling Du","doi":"10.1016/j.synbio.2024.01.005","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.01.005","url":null,"abstract":"<div><p>The biological signaling molecule nitric oxide (NO) has recently emerged as a metabolic precursor for the creation of microbial natural products with diversified structures and biological activities. Within the biosynthetic gene clusters (BGCs) of these compounds, genes associated with NO production pathways have been pinpointed. In this study, we employ a nitric oxide synthase (NOS)-guided genome mining strategy for the targeted discovery of NO-derived bacterial natural products and NO-utilizing biocatalysts. We show that a conserved NOS-containing BGC, distributed across several actinobacterial genomes, is responsible for the biosynthesis of lajollamycin, a unique nitro-tetraene-containing antibiotic whose biosynthetic mechanism remains elusive. Through a combination of in vivo and in vitro studies, we unveil the first cytochrome P450 enzyme capable of catalyzing olefin nitration in natural product biosynthesis. These results not only expand the current knowledge about biosynthetic nitration processes but also offer an efficient way for targeted identification of NO-utilizing metabolic pathways and novel nitrating biocatalysts.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X2400005X/pdfft?md5=a6bd2203eec7a9e46938bd177f8fbe54&pid=1-s2.0-S2405805X2400005X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139504246","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":"Coupled strategy based on regulator manipulation and medium optimization empowers the biosynthetic overproduction of lincomycin","authors":"Xinlu Cai ,&nbsp;Wanlian Xu ,&nbsp;Yang Zheng ,&nbsp;Sendi Wu ,&nbsp;Rundong Zhao ,&nbsp;Nian Wang ,&nbsp;Yaqian Tang ,&nbsp;Meilan Ke ,&nbsp;Qianjin Kang ,&nbsp;Linquan Bai ,&nbsp;Buchang Zhang ,&nbsp;Hang Wu","doi":"10.1016/j.synbio.2024.01.004","DOIUrl":"10.1016/j.synbio.2024.01.004","url":null,"abstract":"<div><p>The biosynthesis of bioactive secondary metabolites, specifically antibiotics, is of great scientific and economic importance. The control of antibiotic production typically involves different processes and molecular mechanism. Despite numerous efforts to improve antibiotic yields, joint engineering strategies for combining genetic manipulation with fermentation optimization remain finite. Lincomycin A (Lin-A), a lincosamide antibiotic, is industrially fermented by <em>Streptomyces lincolnensis</em>. Herein, the leucine-responsive regulatory protein (Lrp)-type regulator SLCG_4846 was confirmed to directly inhibit the lincomycin biosynthesis, whereas indirectly controlled the transcription of <em>SLCG_2919</em>, the first reported repressor in <em>S. lincolnensis</em>. Inactivation of <em>SLCG_4846</em> in the high-yield <em>S. lincolnensis</em> LA219X (LA219XΔ<em>4846</em>) increases the Lin-A production and deletion of <em>SLCG_2919</em> in LA219XΔ<em>4846</em> exhibits superimposed yield increment. Given the effect of the double deletion on cellular primary metabolism of <em>S. lincolnensis</em>, Plackett-Burman design, steepest ascent and response surface methodologies were utilized and employed to optimize the seed medium of this double mutant in shake flask, and Lin-A yield using optimal seed medium was significantly increased over the control. Above strategies were performed in a 15-L fermenter. The maximal yield of Lin-A in LA219XΔ<em>4846-2919</em> reached 6.56 g/L at 216 h, 55.1 % higher than that in LA219X at the parental cultivation (4.23 g/L). This study not only showcases the potential of this strategy to boost lincomycin production, but also could empower the development of high-performance actinomycetes for other antibiotics.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000048/pdfft?md5=446e92fc6eb8a9c8e2f2bf1717276f1a&pid=1-s2.0-S2405805X24000048-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139540290","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":"Efficient biosynthesis of creatine by whole-cell catalysis from guanidinoacetic acid in Corynebacterium glutamicum","authors":"Chunjian Li ,&nbsp;Pengdong Sun ,&nbsp;Guoqing Wei ,&nbsp;Yuqi Zhu ,&nbsp;Jingyuan Li ,&nbsp;Yanfeng Liu ,&nbsp;Jian Chen ,&nbsp;Yang Deng","doi":"10.1016/j.synbio.2024.01.003","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.01.003","url":null,"abstract":"<div><p>Creatine is a naturally occurring derivative of an amino acid commonly utilized in functional foods and pharmaceuticals. Nevertheless, the current industrial synthesis of creatine relies on chemical processes, which may hinder its utilization in certain applications. Therefore, a biological approach was devised that employs whole-cell biocatalysis in the bacterium <em>Corynebacterium glutamicum</em>, which is considered safe for use in food production, to produce safe-for-consumption creatine. The objective of this study was to identify a guanidinoacetate N-methyltransferase (GAMT) with superior catalytic activity for creatine production. Through employing whole-cell biocatalysis, a <em>gamt</em> gene from <em>Mus caroli</em> (<em>Mcgamt</em>) was cloned and expressed in <em>C. glutamicum</em> ATCC 13032, resulting in a creatine titer of 3.37 g/L. Additionally, the study employed a promoter screening strategy that utilized nine native strong promoters in <em>C. glutamicum</em> to enhance the expression level of GAMT. The highest titer was achieved using the P₁₆₇₆ promoter, reaching 4.14 g/L. The conditions of whole-cell biocatalysis were further optimized, resulting in a creatine titer of 5.42 g/L. This is the first report of successful secretory creatine expression in <em>C. glutamicum</em>, which provides a safer and eco-friendly approach for the industrial production of creatine.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000036/pdfft?md5=3a367287745216057893cea4bd134981&pid=1-s2.0-S2405805X24000036-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139434640","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":"Construction of an engineered Escherichia coli for effective synthesis of 2′-fucosyllactose via the salvage pathway","authors":"Shanquan Liang ,&nbsp;Zi He ,&nbsp;Dan Liu ,&nbsp;Shaoqing Yang ,&nbsp;Qiaojuan Yan ,&nbsp;Zhengqiang Jiang","doi":"10.1016/j.synbio.2024.01.001","DOIUrl":"10.1016/j.synbio.2024.01.001","url":null,"abstract":"<div><p>2′-Fucosyllactose (2′-FL) is one of the important functional oligosaccharides in breast milk. So far, few attempts on biosynthesis of 2′-FL by the <em>salvage</em> pathway have been reported. Herein, the <em>salvage</em> pathway enzyme genes were introduced into the <em>E. coli</em> BL21star(DE3) for synthesis of 2′-FL. The 2′-FL titer increased from 1.56 to 2.13 g/L by deleting several endogenous genes on competitive pathways. The α-1,2-fucosyltransferase (WbgL) was selected, and improved the 2′-FL titer to 2.88 g/L. Additionally, the expression level of pathway enzyme genes was tuned through optimizing the plasmid copy number. Furthermore, the spatial distribution of WbgL was enhanced by fusing with the MinD C-tag. After optimizing the fermentation conditions, the 2′-FL titer reached to 7.13 g/L. The final strain produced 59.22 g/L of 2′-FL with 95% molar conversion rate of lactose and 92% molar conversion rate of fucose in a 5 L fermenter. These findings will contribute to construct a highly efficient microbial cell factory to produce 2′-FL or other HMOs.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000012/pdfft?md5=116008fdb513f5cdaaadf0d606a1f43c&pid=1-s2.0-S2405805X24000012-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139392720","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":"Regulatory mechanisms of dopamine metabolism in a marine Meyerozyma guilliermondii GXDK6 under NaCl stress as revealed by integrative multi-omics analysis","authors":"Huijie Sun ,&nbsp;Huashan Bai ,&nbsp;Yonghong Hu ,&nbsp;Sheng He ,&nbsp;Ruihang Wei ,&nbsp;Duotao Meng ,&nbsp;Qiong Jiang ,&nbsp;Hongping Pan ,&nbsp;Peihong Shen ,&nbsp;Qian Ou ,&nbsp;Chengjian Jiang","doi":"10.1016/j.synbio.2024.01.002","DOIUrl":"10.1016/j.synbio.2024.01.002","url":null,"abstract":"<div><p>Dopamine can be used to treat depression, myocardial infarction, and other diseases. However, few reports are available on the de novo microbial synthesis of dopamine from low-cost substrate. In this study, integrated omics technology was used to explore the dopamine metabolism of a novel marine multi-stress-tolerant aromatic yeast <em>Meyerozyma guilliermondii</em> GXDK6. GXDK6 was found to have the ability to biosynthesize dopamine when using glucose as the substrate. 14 key genes for the biosynthesis of dopamine were identified by whole genome-wide analysis. Transcriptomic and proteomic data showed that the expression levels of gene <em>AAT2</em> encoding aspartate aminotransferase (regulating dopamine anabolism) were upregulated, while gene <em>AO-I</em> encoding copper amine oxidase (involved in dopamine catabolism) were downregulated under 10 % NaCl stress compared with non-NaCl stress, thereby contributing to biosynthesis of dopamine. Further, the amount of dopamine under 10 % NaCl stress was 2.51-fold higher than that of zero NaCl, which was consistent with the multi-omics results. Real-time fluorescence quantitative PCR (RT-qPCR) and high-performance liquid chromatography (HPLC) results confirmed the metabolic model of dopamine. Furthermore, by overexpressing <em>AAT2</em>, AST enzyme activity was increased by 24.89 %, the expression of genes related to dopamine metabolism was enhanced, and dopamine production was increased by 56.36 % in recombinant GXDK6AAT2. In conclusion, <em>Meyerozyma guilliermondii</em> GXDK6 could utilize low-cost carbon source to synthesize dopamine, and NaCl stress promoted the biosynthesis of dopamine.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000024/pdfft?md5=e6156a9ebf2f86b0bbe13c1ebc68a027&pid=1-s2.0-S2405805X24000024-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139394088","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":"Good and bad dispositions between archaea and bacteria in the human gut: New insights from metagenomic survey and co-occurrence analysis","authors":"Francesco Candeliere ,&nbsp;Laura Sola ,&nbsp;Stefano Raimondi ,&nbsp;Maddalena Rossi ,&nbsp;Alberto Amaretti","doi":"10.1016/j.synbio.2023.12.007","DOIUrl":"https://doi.org/10.1016/j.synbio.2023.12.007","url":null,"abstract":"<div><p>Archaea are an understudied component of the human microbiome. In this study, the gut archaeome and bacteriome of 60 healthy adults from different region were analyzed by whole-genome shotgun sequencing. Archaea were ubiquitously found in a wide range of abundances, reaching up to 7.2 %. The dominant archaeal phylum was Methanobacteriota, specifically the family Methanobacteriaceae, encompassing more than 50 % of Archaea in 50 samples. The previously underestimated Thermoplasmatota, mostly composed of Methanomassiliicoccaceae, dominated in 10 subjects (&gt;50 %) and was present in all others except one. Halobacteriota, the sole other archaeal phylum, occurred in negligible concentration, except for two samples (4.6–4.8 %). This finding confirmed that the human gut archaeome is primarily composed of methanogenic organisms and among the known methanogenic pathway: i) hydrogenotrophic reduction of CO₂ is the predominant, being the genus <em>Methanobrevibacter</em> and the species <em>Methanobrevibacter smithii</em> the most abundant in the majority of the samples; ii) the second pathway, that involved Methanomassiliicoccales, was the hydrogenotrophic reduction of methyl-compounds; iii) dismutation of acetate or methyl-compounds seemed to be absent. Co-occurrence analysis allowed to unravel correlations between Archaea and Bacteria that shapes the overall structure of the microbial community, allowing to depict a clearer picture of the human gut archaeome.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X2300114X/pdfft?md5=3d1b661b9cc4a1dad608f79e335bb9f8&pid=1-s2.0-S2405805X2300114X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139398957","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":"A toolbox for genetic manipulation in intestinal Clostridium symbiosum","authors":"Pengjie Yang ,&nbsp;Jinzhong Tian ,&nbsp;Lu Zhang ,&nbsp;Hui Zhang ,&nbsp;Gaohua Yang ,&nbsp;Yimeng Ren ,&nbsp;Jingyuan Fang ,&nbsp;Yang Gu ,&nbsp;Weihong Jiang","doi":"10.1016/j.synbio.2023.12.005","DOIUrl":"https://doi.org/10.1016/j.synbio.2023.12.005","url":null,"abstract":"<div><p>Gut microbes are closely related with human health, but remain much to learn. <em>Clostridium symbiosum</em> is a conditionally pathogenic human gut bacterium and regarded as a potential biomarker for early diagnosis of intestinal tumors. However, the absence of an efficient toolbox that allows diverse genetic manipulations of this bacterium limits its in-depth studies. Here, we obtained the complete genome sequence of <em>C. symbiosum</em> ATCC 14940, a representative strain of <em>C. symbiosum</em>. On this basis, we further developed a series of genetic manipulation methods for this bacterium. Firstly, following the identification of a functional replicon pBP1 in <em>C. symbiosum</em> ATCC 14940, a highly efficient conjugative DNA transfer method was established, enabling the rapid introduction of exogenous plasmids into cells. Next, we constructed a dual-plasmid CRISPR/Cas12a system for genome editing in this bacterium, reaching over 60 % repression for most of the chosen genes as well as efficient deletion (&gt;90 %) of three target genes. Finally, this toolbox was used for the identification of crucial functional genes, involving growth, synthesis of important metabolites, and virulence of <em>C. symbiosum</em> ATCC 14940. Our work has effectively established and optimized genome editing methods in intestinal <em>C. symbiosum</em>, thereby providing strong support for further basic and application research in this bacterium.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2023-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X23001126/pdfft?md5=4bbc1e472ee3d742a0ea74d0155338f8&pid=1-s2.0-S2405805X23001126-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139090289","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":"Towards a hybrid model-driven platform based on flux balance analysis and a machine learning pipeline for biosystem design","authors":"Debiao Wu ,&nbsp;Feng Xu ,&nbsp;Yaying Xu,&nbsp;Mingzhi Huang,&nbsp;Zhimin Li,&nbsp;Ju Chu","doi":"10.1016/j.synbio.2023.12.004","DOIUrl":"https://doi.org/10.1016/j.synbio.2023.12.004","url":null,"abstract":"<div><p>Metabolic modeling and machine learning (ML) are crucial components of the evolving next-generation tools in systems and synthetic biology, aiming to unravel the intricate relationship between genotype, phenotype, and the environment. Nonetheless, the comprehensive exploration of integrating these two frameworks, and fully harnessing the potential of fluxomic data, remains an unexplored territory. In this study, we present, rigorously evaluate, and compare ML-based techniques for data integration. The hybrid model revealed that the overexpression of six target genes and the knockout of seven target genes contribute to enhanced ethanol production. Specifically, we investigated the influence of succinate dehydrogenase (SDH) on ethanol biosynthesis in <em>Saccharomyces cerevisiae</em> through shake flask experiments. The findings indicate a noticeable increase in ethanol yield, ranging from 6 % to 10 %, in SDH subunit gene knockout strains compared to the wild-type strain. Moreover, in pursuit of a high-yielding strain for ethanol production, dual-gene deletion experiments were conducted targeting glycerol-3-phosphate dehydrogenase (GPD) and SDH. The results unequivocally demonstrate significant enhancements in ethanol production for the engineered strains <em>Δsdh</em>4<em>Δgpd</em>1, <em>Δsdh</em>5<em>Δgpd</em>1, <em>Δsdh</em>6<em>Δgpd</em>1, <em>Δsdh</em>4<em>Δgpd</em>2, <em>Δsdh</em>5<em>Δgpd</em>2, and <em>Δsdh</em>6<em>Δgpd</em>2, with improvements of 21.6 %, 27.9 %, and 22.7 %, respectively. Overall, the results highlighted that integrating mechanistic flux features substantially improves the prediction of gene knockout strains not accounted for in metabolic reconstructions. In addition, the finding in this study delivers valuable tools for comprehending and manipulating intricate phenotypes, thereby enhancing prediction accuracy and facilitating deeper insights into mechanistic aspects within the field of synthetic biology.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2023-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X23001114/pdfft?md5=90a292087c9fa4adcf87d4ad13c22daf&pid=1-s2.0-S2405805X23001114-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139090288","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":"A flexible, modular and versatile functional part assembly toolkit for gene cluster engineering in Streptomyces","authors":"Xuejin Zhao ,&nbsp;Yeqing Zong ,&nbsp;Qiuli Lou ,&nbsp;Chenrui Qin ,&nbsp;Chunbo Lou","doi":"10.1016/j.synbio.2023.12.003","DOIUrl":"https://doi.org/10.1016/j.synbio.2023.12.003","url":null,"abstract":"<div><p><em>Streptomyces</em> has enormous potential to produce novel natural products (NPs) as it harbors a huge reservoir of uncharacterized and silent natural product biosynthetic gene clusters (BGCs). However, the lack of efficient gene cluster engineering strategies has hampered the pace of new drug discovery. Here, we developed an easy-to-use, highly flexible DNA assembly toolkit for gene cluster engineering. The DNA assembly toolkit is compatible with various DNA assembling approaches including Biobrick, Golden Gate, CATCH, yeast homologous recombination-based DNA assembly and homing endonuclease-mediated assembly. This compatibility offers great flexibility in handling multiple genetic parts or refactoring large gene clusters. To demonstrate the utility of this toolkit, we quantified a library of modular regulatory parts, and engineered a gene cluster (<em>act</em>) using characterized promoters that led to increased production. Overall, this work provides a powerful part assembly toolkit that can be used for natural product discovery and optimization in <em>Streptomyces</em>.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2023-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X23001102/pdfft?md5=c4cdd2cc2eada7bc6af8f878d3c1d487&pid=1-s2.0-S2405805X23001102-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139107543","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}