Synthetic and Systems Biotechnology最新文献

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Heterologous activation and metabolites identification of the pks7 gene cluster from Saccharopolyspora erythraea 红球藻 pks7 基因簇的异源激活和代谢物鉴定
IF 4.4 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-05-09 DOI: 10.1016/j.synbio.2024.05.004
Hao Tang , Xingchi Yang , Wenzong Wang , Xingjun Cui , Wenping Wei , Jing Wu , Peng Sun , Bang-Ce Ye
{"title":"Heterologous activation and metabolites identification of the pks7 gene cluster from Saccharopolyspora erythraea","authors":"Hao Tang ,&nbsp;Xingchi Yang ,&nbsp;Wenzong Wang ,&nbsp;Xingjun Cui ,&nbsp;Wenping Wei ,&nbsp;Jing Wu ,&nbsp;Peng Sun ,&nbsp;Bang-Ce Ye","doi":"10.1016/j.synbio.2024.05.004","DOIUrl":"10.1016/j.synbio.2024.05.004","url":null,"abstract":"<div><p>The microbial genome remains a huge treasure trove for the discovery of diverse natural products. <em>Saccharopolyspora erythraea</em> NRRL23338, the industry producer of erythromycin, has a dozen of biosynthetic gene clusters whose encoding products are unidentified. Heterologous expression of one of the polyketide clusters <em>pks7</em> in <em>Streptomyces albus</em> B4 chassis resulted in the characterization of its function responsible for synthesizing both 6-methylsalicyclic acid and 6-ethylsalicyclic acid. Meanwhile, two new 6-ethylsalicyclic acid ester derivatives were isolated as shunt metabolites. Their structures were identified by comprehensive analysis of MS and NMR experiments. Putative functions of genes within the <em>pks7</em> BGC were also discussed.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000723/pdfft?md5=af1e81459f9bf129dd7836ed458efcf4&pid=1-s2.0-S2405805X24000723-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141043211","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
Rational construction of genome-minimized Streptomyces host for the expression of secondary metabolite gene clusters 合理构建基因组最小化链霉菌宿主以表达次生代谢物基因簇
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-05-05 DOI: 10.1016/j.synbio.2024.04.017
Hui Li , Sheng Gao , Sanyuan Shi , Xiaomin Zhao , Haoyu Ye , Yunzi Luo
{"title":"Rational construction of genome-minimized Streptomyces host for the expression of secondary metabolite gene clusters","authors":"Hui Li ,&nbsp;Sheng Gao ,&nbsp;Sanyuan Shi ,&nbsp;Xiaomin Zhao ,&nbsp;Haoyu Ye ,&nbsp;Yunzi Luo","doi":"10.1016/j.synbio.2024.04.017","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.017","url":null,"abstract":"<div><p><em>Streptomyces</em> offer a wealth of naturally occurring compounds with diverse structures, many of which possess significant pharmaceutical values. However, new product exploration and increased yield of specific compounds in <em>Streptomyces</em> have been technically challenging due to their slow growth rate, complex culture conditions and intricate genetic backgrounds. In this study, we screened dozens of <em>Streptomyces</em> strains inhabiting in a plant rhizosphere for fast-growing candidates, and further employed CRISPR/Cas-based engineering techniques for stepwise refinement of a particular strain, <em>Streptomyces</em> sp. A-14 that harbors a 7.47 Mb genome. After strategic removal of nonessential genomic regions and most gene clusters, we reduced its genome size to 6.13 Mb, while preserving its growth rate to the greatest extent. We further demonstrated that cleaner metabolic background of this engineered strain was well suited for the expression and characterization of heterologous gene clusters, including the biosynthetic pathways of actinorhodin and polycyclic tetramate macrolactams. Moreover, this streamlined genome is anticipated to facilitate directing the metabolic flux towards the production of desired compounds and increasing their yields.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000681/pdfft?md5=96b78023849d6969617af7fe24b33ed0&pid=1-s2.0-S2405805X24000681-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140901750","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
Simultaneous Multiplex Genome Loci Editing of Halomonas bluephagenesis Using an Engineered CRISPR-guided Base Editor 使用工程化 CRISPR 引导的碱基编辑器同时对蓝光单胞菌进行多重基因组位点编辑
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-25 DOI: 10.1016/j.synbio.2024.04.016
Yulin Zhang , Yang Zheng , Qiwen Hu , Zhen Hu , Jiyuan Sun , Ping Cheng , Xiancai Rao , Xiao-Ran Jiang
{"title":"Simultaneous Multiplex Genome Loci Editing of Halomonas bluephagenesis Using an Engineered CRISPR-guided Base Editor","authors":"Yulin Zhang ,&nbsp;Yang Zheng ,&nbsp;Qiwen Hu ,&nbsp;Zhen Hu ,&nbsp;Jiyuan Sun ,&nbsp;Ping Cheng ,&nbsp;Xiancai Rao ,&nbsp;Xiao-Ran Jiang","doi":"10.1016/j.synbio.2024.04.016","DOIUrl":"10.1016/j.synbio.2024.04.016","url":null,"abstract":"<div><p><em>Halomonas bluephagenesis</em> TD serves as an exceptional chassis for next generation industrial biotechnology to produce various products. However, the simultaneous editing of multiple loci in <em>H. bluephagenesis</em> TD remains a significant challenge. Herein, we report the development of a multiple loci genome editing system, named CRISPR-deaminase-assisted base editor (CRISPR-BE) in <em>H. bluephagenesis</em> TD. This system comprises two components: a cytidine (CRISPR-cBE) and an adenosine (CRISPR-aBE) deaminase-based base editor. CRISPR-cBE can introduce a cytidine to thymidine mutation with an efficiency of up to 100% within a 7-nt editing window in <em>H. bluephagenesis</em> TD. Similarly, CRISPR-aBE demonstrates an efficiency of up to 100% in converting adenosine to guanosine mutation within a 7-nt editing window. CRISPR-cBE has been further validated and successfully employed for simultaneous multiplexed editing in <em>H. bluephagenesis</em> TD. Our findings reveal that CRISPR-cBE efficiently inactivated all six copies of the IS1086 gene simultaneously by introducing stop codon. This system achieved an editing efficiency of 100% and 41.67% in inactivating two genes and three genes, respectively. By substituting the P<sub>cas</sub> promoter with the inducible promoter P<sub>Mmp1</sub>, we optimized CRISPR-cBE system and ultimately achieved 100% editing efficiency in inactivating three genes. In conclusion, our research offers a robust and efficient method for concurrently modifying multiple loci in <em>H. bluephagenesis</em> TD, opening up vast possibilities for industrial applications in the future.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X2400067X/pdfft?md5=b15c2dd720390fb769b69b577cbe14b6&pid=1-s2.0-S2405805X2400067X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140785797","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 a non-model yeast Rhodotorula mucilaginosa for terpenoids synthesis 改造非模式酵母 Rhodotorula mucilaginosa 以合成萜类化合物
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-23 DOI: 10.1016/j.synbio.2024.04.015
Qiongqiong Chen , Liting Lyu , Haizhao Xue , Aabid Manzoor Shah , Zongbao Kent Zhao
{"title":"Engineering a non-model yeast Rhodotorula mucilaginosa for terpenoids synthesis","authors":"Qiongqiong Chen ,&nbsp;Liting Lyu ,&nbsp;Haizhao Xue ,&nbsp;Aabid Manzoor Shah ,&nbsp;Zongbao Kent Zhao","doi":"10.1016/j.synbio.2024.04.015","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.015","url":null,"abstract":"<div><p>Terpenoids have tremendous biological activities and are widely employed in food, healthcare and pharmaceutical industries. Using synthetic biology to product terpenoids from microbial cell factories presents a promising alternative route compared to conventional methods such as chemical synthesis or phytoextraction. The red yeast <em>Rhodotorula mucilaginosa</em> has been widely studied due to its natural production capacity of carotenoid and lipids, indicating a strong endogenous isoprene pathway with readily available metabolic intermediates. This study constructed several engineered strains of <em>R. mucilaginosa</em> with the aim of producing different terpenoids. Monoterpene α-terpineol was produced by expressing the α-terpineol synthase from <em>Vitis vinifera</em>. The titer of α-terpineol was further enhanced to 0.39 mg/L by overexpressing the endogenous rate-limiting gene of the MVA pathway. Overexpression of α-farnesene synthase from <em>Malus domestica,</em> in combination with MVA pathway rate-limiting gene resulted in significant increase in α-farnesene production, reaching a titer of 822 mg/L. The carotenoid degradation product β-ionone was produced at a titer of 0.87 mg/L by expressing the β-ionone synthase from <em>Petunia hybrida</em>. This study demonstrates the potential of <em>R. mucilaginosa</em> as a platform host for the direct biosynthesis of various terpenoids and provides insights for further development of such platforms.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000668/pdfft?md5=59f13b657d4fdba39f6b86f484b19421&pid=1-s2.0-S2405805X24000668-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140644175","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 the activity and succinyl-CoA specificity of 3-ketoacyl-CoA thiolase Tfu_0875 through rational binding pocket engineering 通过合理的结合口袋工程提高 3-酮酰-CoA硫醇酶 Tfu_0875 的活性和琥珀酰-CoA 特异性
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-20 DOI: 10.1016/j.synbio.2024.04.014
Lixia Liu , Shuang Liu , Xiangyang Hu , Shenghu Zhou , Yu Deng
{"title":"Enhancing the activity and succinyl-CoA specificity of 3-ketoacyl-CoA thiolase Tfu_0875 through rational binding pocket engineering","authors":"Lixia Liu ,&nbsp;Shuang Liu ,&nbsp;Xiangyang Hu ,&nbsp;Shenghu Zhou ,&nbsp;Yu Deng","doi":"10.1016/j.synbio.2024.04.014","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.014","url":null,"abstract":"<div><p>The 3-ketoacyl-CoA thiolase is the rate-limiting enzyme for linear dicarboxylic acids production. However, the promiscuous substrate specificity and suboptimal catalytic performance have restricted its application. Here we present both biochemical and structural analyses of a high-efficiency 3-ketoacyl-CoA thiolase Tfu_0875. Notably, Tfu_0875 displayed heightened activity and substrate specificity for succinyl-CoA, a key precursor in adipic acid production. To enhance its performance, a deep learning approach (DLKcat) was employed to identify effective mutants, and a computational strategy, known as the greedy accumulated strategy for protein engineering (GRAPE), was used to accumulate these effective mutants. Among the mutants, Tfu_0875<sup>N249W/L163H/E217L</sup> exhibited the highest specific activity (320% of wild-type Tfu_0875), the greatest catalytic efficiency (<em>k</em><sub><em>cat</em></sub>/<em>K</em><sub><em>M</em></sub> = 1.00 min<sup>−1</sup>mM<sup>−1</sup>), the highest succinyl-CoA specificity (<em>K</em><sub><em>M</em></sub> = 0.59 mM, 28.1% of Tfu_0875) and dramatically reduced substrate binding energy (−30.25 kcal mol<sup>−1</sup> <em>v.s.</em> −15.94 kcal mol<sup>−1</sup>). A structural comparison between Tfu_0875<sup>N249W/L163H/E217L</sup> and the wild type Tfu_0875 revealed that the increased interaction between the enzyme and succinyl-CoA was the primary reason for the enhanced enzyme activity. This interaction facilitated rapid substrate anchoring and stabilization. Furthermore, a reduced binding pocket volume improved substrate specificity by enhancing the complementarity between the binding pocket and the substrate in stereo conformation. Finally, our rationally designed mutant, Tfu_0875<sup>N249W/L163H/E217L</sup>, increased the adipic acid titer by 1.35-fold compared to the wild type Tfu_0875 in shake flask. The demonstrated enzymatic methods provide a promising enzyme variant for the adipic acid production. The above effective substrate binding pocket engineering strategy can be beneficial for the production of other industrially competitive biobased chemicals when be applied to other thiolases.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000656/pdfft?md5=8fc201d89c4bea2c36280d80ceb2cb5d&pid=1-s2.0-S2405805X24000656-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140638706","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
Rational engineering of homospermidine synthase for enhanced catalytic efficiency toward spermidine synthesis 对同胚乳苷合成酶进行合理工程改造,提高其对精胺合成的催化效率
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-19 DOI: 10.1016/j.synbio.2024.04.012
Wenjing Liu, Xiaoxiang Hu, Yi Yan, Yujie Cai
{"title":"Rational engineering of homospermidine synthase for enhanced catalytic efficiency toward spermidine synthesis","authors":"Wenjing Liu,&nbsp;Xiaoxiang Hu,&nbsp;Yi Yan,&nbsp;Yujie Cai","doi":"10.1016/j.synbio.2024.04.012","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.012","url":null,"abstract":"<div><p>Spermidine is a naturally occurring polyamine widely utilized in the prevention and treatment of various diseases. Current spermidine biosynthetic methods have problems such as low efficiency and complex multi-enzyme catalysis. Based on sequence-structure-function relationships, we engineered the widely studied homospermidine synthase from <em>Blastochloris viridis</em> (<em>Bv</em>HSS) and obtained mutants that could catalyze the production of spermidine from 1,3-diaminopropane and putrescine. The specific activities of <em>Bv</em>HSS and the mutants D361E and E232D + D361E (E232D-D) were 8.72, 46.04 and 48.30 U/mg, respectively. The optimal pH for both mutants was 9.0, and the optimal temperature was 50 °C. Molecular docking and dynamics simulations revealed that mutating aspartic acid at position 361 to glutamic acid narrowed the substrate binding pocket, promoting stable spermidine production. Conversely, mutating glutamic acid at position 232 to aspartic acid enlarged the substrate channel entrance, facilitating substrate entry into the active pocket and enhancing spermidine generation. In whole-cell catalysis lasting 6 h, D361E and E232D-D synthesized 725.3 and 933.5 mg/L of spermidine, respectively. This study offers a practical approach for single-enzyme catalyzed spermidine synthesis and sheds light on the crucial residues influencing homospermidine synthase catalytic activity in spermidine production.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000632/pdfft?md5=6a16965eec979bf54901fb3e492828c8&pid=1-s2.0-S2405805X24000632-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140632556","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
SemiSynBio: A new era for neuromorphic computing SemiSynBio:神经形态计算的新时代
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-18 DOI: 10.1016/j.synbio.2024.04.013
Ruicun Liu , Tuoyu Liu , Wuge Liu , Boyu Luo , Yuchen Li , Xinyue Fan , Xianchao Zhang , Wei Cui , Yue Teng
{"title":"SemiSynBio: A new era for neuromorphic computing","authors":"Ruicun Liu ,&nbsp;Tuoyu Liu ,&nbsp;Wuge Liu ,&nbsp;Boyu Luo ,&nbsp;Yuchen Li ,&nbsp;Xinyue Fan ,&nbsp;Xianchao Zhang ,&nbsp;Wei Cui ,&nbsp;Yue Teng","doi":"10.1016/j.synbio.2024.04.013","DOIUrl":"10.1016/j.synbio.2024.04.013","url":null,"abstract":"<div><p>Neuromorphic computing has the potential to achieve the requirements of the next-generation artificial intelligence (AI) systems, due to its advantages of adaptive learning and parallel computing. Meanwhile, biocomputing has seen ongoing development with the rise of synthetic biology, becoming the driving force for new generation semiconductor synthetic biology (SemiSynBio) technologies. DNA-based biomolecules could potentially perform the functions of Boolean operators as logic gates and be used to construct artificial neural networks (ANNs), providing the possibility of executing neuromorphic computing at the molecular level. Herein, we briefly outline the principles of neuromorphic computing, describe the advances in DNA computing with a focus on synthetic neuromorphic computing, and summarize the major challenges and prospects for synthetic neuromorphic computing. We believe that constructing such synthetic neuromorphic circuits will be an important step toward realizing neuromorphic computing, which would be of widespread use in biocomputing, DNA storage, information security, and national defense.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000644/pdfft?md5=1e4605b2d76880602fd20da7183a1007&pid=1-s2.0-S2405805X24000644-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140791740","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
Site-directed mutagenesis of bifunctional riboflavin kinase/FMN adenylyltransferase via CRISPR/Cas9 to enhance riboflavin production 通过 CRISPR/Cas9 对双功能核黄素激酶/FMN 腺苷酸转移酶进行定点诱变,提高核黄素产量
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-16 DOI: 10.1016/j.synbio.2024.04.011
Bing Fu , Meng Chen , Xianfeng Bao , Jiajie Lu , Zhiwen Zhu , Fuyao Guan , Chuyang Yan , Peize Wang , Linglin Fu , Ping Yu
{"title":"Site-directed mutagenesis of bifunctional riboflavin kinase/FMN adenylyltransferase via CRISPR/Cas9 to enhance riboflavin production","authors":"Bing Fu ,&nbsp;Meng Chen ,&nbsp;Xianfeng Bao ,&nbsp;Jiajie Lu ,&nbsp;Zhiwen Zhu ,&nbsp;Fuyao Guan ,&nbsp;Chuyang Yan ,&nbsp;Peize Wang ,&nbsp;Linglin Fu ,&nbsp;Ping Yu","doi":"10.1016/j.synbio.2024.04.011","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.011","url":null,"abstract":"<div><p>Vitamin B<sub>2</sub> is an essential water-soluble vitamin. For most prokaryotes, a bifunctional enzyme called FAD synthase catalyzes the successive conversion of riboflavin to FMN and FAD. In this study, the plasmid pNEW-AZ containing six key genes for the riboflavin synthesis was transformed into strain R2 with the deleted FMN riboswitch, yielding strain R5. The R5 strain could produce 540.23 ± 5.40 mg/L riboflavin, which was 10.61 % higher than the R4 strain containing plasmids pET-AE and pAC-Z harboring six key genes. To further enhance the production of riboflavin, homology matching and molecular docking were performed to identify key amino acid residues of FAD synthase. Nine point mutation sites were identified. By comparing riboflavin kinase activity, mutations of T203D and N210D, which respectively decreased by 29.90 % and 89.32 % compared to wild-type FAD synthase, were selected for CRISPR/Cas9 gene editing of the genome, generating engineered strains R203 and R210. pNEW-AZ was transformed into R203, generating R6. R6 produced 657.38 ± 47.48 mg/L riboflavin, a 21.69 % increase compared to R5. This study contributes to the high production of riboflavin in recombinant <em>E. coli</em> BL21.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000620/pdfft?md5=a81bfa53af322180bb704e3bb42e123e&pid=1-s2.0-S2405805X24000620-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140558895","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
Facilitating stable gene integration expression and copy number amplification in Bacillus subtilis through a reversible homologous recombination switch 通过可逆同源重组开关促进枯草芽孢杆菌中稳定的基因整合表达和拷贝数扩增
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-16 DOI: 10.1016/j.synbio.2024.04.010
Haoyu Guo , Rongzhen Tian , Yaokang Wu , Xueqin Lv , Jianghua Li , Long Liu , Guocheng Du , Jian Chen , Yanfeng Liu
{"title":"Facilitating stable gene integration expression and copy number amplification in Bacillus subtilis through a reversible homologous recombination switch","authors":"Haoyu Guo ,&nbsp;Rongzhen Tian ,&nbsp;Yaokang Wu ,&nbsp;Xueqin Lv ,&nbsp;Jianghua Li ,&nbsp;Long Liu ,&nbsp;Guocheng Du ,&nbsp;Jian Chen ,&nbsp;Yanfeng Liu","doi":"10.1016/j.synbio.2024.04.010","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.010","url":null,"abstract":"<div><p>Strengthening the expression level of integrated genes on the genome is crucial for consistently expressing key enzymes in microbial cell factories for efficient bioproduction in synthetic biology. In comparison to plasmid-based multi-copy expression, the utilization of chromosomal multi-copy genes offers increased stability of expression level, diminishes the metabolic burden on host cells, and enhances overall genetic stability. In this study, we developed the “<strong>BacAmp</strong>”, a stabilized gene integration expression and copy number amplification system for high-level expression in <em>Bacillus subtilis</em>, which was achieved by employing a combination of repressor and non-natural amino acids (ncAA)-dependent expression system to create a reversible switch to control the key gene <em>recA</em> for homologous recombination. When the reversible switch is turned on, genome editing and gene amplification can be achieved. Subsequently, the reversible switch was turned off therefore stabilizing the gene copy number. The stabilized gene amplification system marked by green fluorescent protein, achieved a 3-fold increase in gene expression by gene amplification and maintained the average gene copy number at 10 after 110 generations. When we implemented the gene amplification system for the regulation of <em>N</em>-acetylneuraminic acid (NeuAc) synthesis, the copy number of the critical gene increased to an average of 7.7, which yielded a 1.3-fold NeuAc titer. Our research provides a new avenue for gene expression in synthetic biology and can be applied in metabolic engineering in <em>B. subtilis</em>.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000619/pdfft?md5=3dfe16dd56984009fb82ba6dd2d3bbcc&pid=1-s2.0-S2405805X24000619-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140650099","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
Filamentous morphology engineering of bacteria by iron metabolism modulation through MagR expression 通过表达 MagR 调节铁代谢,实现细菌的丝状形态工程
IF 4.8 2区 生物学
Synthetic and Systems Biotechnology Pub Date : 2024-04-15 DOI: 10.1016/j.synbio.2024.04.009
Mengke Wei , Chenyang Han , Xiujuan Zhou , Tianyang Tong , Jing Zhang , Xinmiao Ji , Peng Zhang , Yanqi Zhang , Yan Liu , Xin Zhang , Tiantian Cai , Can Xie
{"title":"Filamentous morphology engineering of bacteria by iron metabolism modulation through MagR expression","authors":"Mengke Wei ,&nbsp;Chenyang Han ,&nbsp;Xiujuan Zhou ,&nbsp;Tianyang Tong ,&nbsp;Jing Zhang ,&nbsp;Xinmiao Ji ,&nbsp;Peng Zhang ,&nbsp;Yanqi Zhang ,&nbsp;Yan Liu ,&nbsp;Xin Zhang ,&nbsp;Tiantian Cai ,&nbsp;Can Xie","doi":"10.1016/j.synbio.2024.04.009","DOIUrl":"https://doi.org/10.1016/j.synbio.2024.04.009","url":null,"abstract":"<div><p>The morphology is the consequence of evolution and adaptation. <em>Escherichia coli</em> is rod-shaped bacillus with regular dimension of about 1.5 μm long and 0.5 μm wide. Many shape-related genes have been identified and used in morphology engineering of this bacteria. However, little is known about if specific metabolism and metal irons could modulate bacteria morphology. Here in this study, we discovered filamentous shape change of <em>E. coli</em> cells overexpressing pigeon MagR, a putative magnetoreceptor and extremely conserved iron-sulfur protein. Comparative transcriptomic analysis strongly suggested that the iron metabolism change and iron accumulation due to the overproduction of MagR was the key to the morphological change. This model was further validated, and filamentous morphological change was also achieved by supplement <em>E. coli</em> cells with iron in culture medium or by increase the iron uptake genes such as entB and fepA. Our study extended our understanding of morphology regulation of bacteria, and may also serves as a prototype of morphology engineering by modulating the iron metabolism.</p></div>","PeriodicalId":22148,"journal":{"name":"Synthetic and Systems Biotechnology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405805X24000607/pdfft?md5=fbf82cb73481e05cbbd6201f98ba4cb2&pid=1-s2.0-S2405805X24000607-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140605106","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}
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