Sheng wu gong cheng xue bao = Chinese journal of biotechnology最新文献_第7页

[Efficient synthesis of polydatin by a two-enzyme coupled with one-pot method].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240398

Jingli Dai, Zixu Yan, Kexue Zhao, Xiaoli Li, Yongjun Zang, Qilin Xu, Fucheng Zhu

{"title":"[Efficient synthesis of polydatin by a two-enzyme coupled with one-pot method].","authors":"Jingli Dai, Zixu Yan, Kexue Zhao, Xiaoli Li, Yongjun Zang, Qilin Xu, Fucheng Zhu","doi":"10.13345/j.cjb.240398","DOIUrl":"https://doi.org/10.13345/j.cjb.240398","url":null,"abstract":"Traditional Chinese medicine of Polygonum cuspidatum has been utilized in China for thousands of years. Its primary active compound, polydatin, exhibits a variety of pharmacological effects including the regulation of glucose and lipid metabolism, suppression of cough and asthma, as well as antibacterial and anti-inflammatory properties. However, conventional methods for polydatin production are inadequate to satisfy the market demand. This study aims to explore the green and efficient preparation of polydatin. With resveratrol as the substrate, we efficiently synthesized polydatin by using the triple mutant IGW (Y14I/I62G/M315W) of the glycosyltransferase UGTBS based on a strategy of two-enzyme coupled with one-pot and realized the recycling of uridine diphosphate-glucose (UDPG). The conditions of the two-enzyme reaction were optimized. Under the conditions of 35 ℃, pH 8.0, IGW: AtSuSy1 activity ratio of 3:4, dimethyl sulfoxide (DMSO) volume fraction of 5%, uridine diphosphate (UDP) concentration of 0.10 mmol/L, and sucrose concentration of 0.6 mol/L, the conversion of 2 mmol/L resveratrol reached 80.6% within 1 h, and the proportion of polydatin was over 90%. This study achieved the recycling of UDPG via a two-enzyme coupling system and shortened the reaction time. At the same time, the fed-batch strategy was adopted, and the yield of polydatin reached 6.28 g/L after 24 h in the one-pot coupling reaction, which provided a new strategy for green and efficient preparation of polydatin.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"461-473"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[A novel carbonyl reductase for the synthesis of (R)-tolvaptan].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240238

Yahui Liu, Xuming Wang, Shuo Ma, Keyu Liu, Wei Li, Lulu Zhang, Jie DU, Honglei Zhang

{"title":"[A novel carbonyl reductase for the synthesis of (R)-tolvaptan].","authors":"Yahui Liu, Xuming Wang, Shuo Ma, Keyu Liu, Wei Li, Lulu Zhang, Jie DU, Honglei Zhang","doi":"10.13345/j.cjb.240238","DOIUrl":"https://doi.org/10.13345/j.cjb.240238","url":null,"abstract":"Screening carbonyl reductases with the ability to catalyze the reduction of complex carbonyl compounds is of great significance for the biosynthesis of R-tolvaptan(R-TVP). In this study, the target carbonyl reductase in the crude enzyme extract of rabbit liver was separated, purified, and identified by ammonium sulfate precipitation, gel-filtration chromatography, ion exchange chromatography, affinity chromatography, and protein mass spectrometry. With the rabbit liver genome as the template, the gene encoding the carbonyl reductase rlsr5 was amplified by PCR and the recombinant strain was successfully constructed. After RLSR5 was purified by affinity chromatography, its enzymatic properties were characterized. The results indicated that the gene sequence of rlsr5 was 972 bp, encoding a protein with a molecular weight of 40 kDa. RLSR5 was a dimeric protein, and each monomer was composed of a (α/β)8-barrel structure. RLSR5 could asymmetrically reduce 7-chloro-1-[2-methyl-4-[(2- methylbenzoyl)amino]benzoyl]-5-oxo-2,3,4,5-tetrahydro-1H-1-benzazepine (prochiral ketone, PK) to synthesize R-TVP. The specific activity of the enzyme was 36.64 U/mg, and the optical purity of the product was 99%. This enzyme showcased the optimal performance at pH 6.0 and 30 °C. It was independent of metal ions, with the activity enhanced by Mn2+. This study lays a foundation for the biosynthesis of tolvaptan of optical grade.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"321-332"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041813","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Application of biomanufacturing in polymer flooding].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240257

Junping Zhou, Qilu Pan, Lianggang Huang, Kan Zhan, Heng Tang, Liqun Jin, Yuguo Zheng

{"title":"[Application of biomanufacturing in polymer flooding].","authors":"Junping Zhou, Qilu Pan, Lianggang Huang, Kan Zhan, Heng Tang, Liqun Jin, Yuguo Zheng","doi":"10.13345/j.cjb.240257","DOIUrl":"https://doi.org/10.13345/j.cjb.240257","url":null,"abstract":"In China, the crude oil supply is highly dependent on overseas countries, and thus strengthening crude oil self-sufficiency has become an important issue of the national energy security. Tertiary oil recovery, especially polymer flooding, has been widely applied in large oil fields in China, which can increase the recovery rate by 15%-20% compared with water flooding. However, the widely used oil flooding polymers show poor thermal stability and salinity tolerance, complicated synthesis ways of monomers, and environmental unfriendliness. Moreover, the polymer flooding induces problems including pore plugging, heterogeneity intensification, high dispersion of remaining oil resources, pressure rise in injection wells, and low efficiency circulation of injection medium, which restrict the subsequent recovery of old oil fields. Here, we systematically review the developing and current situations of polymer flooding, introduce the innovative biomanufacturing of oil flooding polymers and their monomers or precursors as well as low-cost bio-based chemical raw materials for multiple compound flooding. The comprehensive study of the relationships between microbial fermentation metabolites and polymer flooding will reveal the green and low-carbon paths for polymer flooding. Such study will enable the application of enzymes produced by microorganisms in polymer production and polymer plugging removal after polymer flooding as well as the application of microbial metabolites such as biosurfactants, organic acids, alcohols, biogas, and amino acids in enhancing oil recovery. This review suggests that incorporating biomanufacturing into polymer flooding will ensure the high productivity and stability for crude oil production in China.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"148-172"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[An efficient assembly method for a viral genome based on T7 endonuclease Ⅰ-mediated error correction].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240302

Xuwei Zhang, Bin Wen, Fei Wang, Xuejun Wang, Liyan Liu, Shumei Wang, Shengqi Wang

{"title":"[An efficient assembly method for a viral genome based on T7 endonuclease Ⅰ-mediated error correction].","authors":"Xuwei Zhang, Bin Wen, Fei Wang, Xuejun Wang, Liyan Liu, Shumei Wang, Shengqi Wang","doi":"10.13345/j.cjb.240302","DOIUrl":"https://doi.org/10.13345/j.cjb.240302","url":null,"abstract":"Gene synthesis is an enabling technology that supports the development of synthetic biology. The existing approaches for de novo gene synthesis generally have tedious operation, low efficiency, high error rates, and limited product lengths, being difficult to support the huge demand of synthetic biology. The assembly and error correction are the keys in gene synthesis. This study first designed the oligonucleotide sequences by reasonably splitting the virus genome of approximately 10 kb by balancing the parameters of sequence design software ability, PCR amplification ability, and assembly enzyme assembly ability. Then, two-step PCR was performed with high-fidelity polymerase to complete the de novo synthesis of 3.0 kb DNA fragments, and error correction reactions were performed with T7 endonuclease Ⅰ for the products from different stages of PCR. Finally, the virus genome was assembled by 3.0 kb DNA fragments from de novo synthesis and error correction and then sequenced. The experimental results showed that the proposed method successfully produced the DNA fragment of about 10 kb and reduced the probability of large fragment mutations during the assembly process, with the lowest error rate reaching 0.36 errors/kb. In summary, this study developed an efficient de novo method for synthesizing a viral genome of about 10 kb with T7 endonuclease Ⅰ-mediated error correction. This method enabled the synthesis of a 10 kb viral genome in one day and the correct plasmid of the viral genome in five days. This study optimized the de novo gene synthesis process, reduced the error rate, simplified the synthesis and assembly steps, and reduced the cost of viral genome assembly.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"385-396"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Enzymatic MBH reaction catalyzed by an artificial enzyme designed with the introduction of an unnatural tertiary amine cofactor].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240223

Ya Wei, Chongwen Chen, Yingjia Tong, Zhi Zhou

引用次数: 0

[Mining, characterization, and expression of a fructan sucrase for efficient conversion of soybean oligosaccharides].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240362

Bin Wang, Jingru Ying, Yuanyuan Chen, Zemin Fang, Yazhong Xiao, Wei Fang, Dongbang Yao

{"title":"[Mining, characterization, and expression of a fructan sucrase for efficient conversion of soybean oligosaccharides].","authors":"Bin Wang, Jingru Ying, Yuanyuan Chen, Zemin Fang, Yazhong Xiao, Wei Fang, Dongbang Yao","doi":"10.13345/j.cjb.240362","DOIUrl":"https://doi.org/10.13345/j.cjb.240362","url":null,"abstract":"The high content of sucrose and raffinose reduces the prebiotic value of soybean oligosaccharides. Fructan sucrases can catalyze the conversion of sucrose and raffinose to high-value products such as fructooligosaccharides and melibiose. To obtain a fructan sucrase that can efficiently convert soybean oligosaccharides, we first mined the fructan sucrase gene from microorganisms in the coastal areas of Xisha Islands and Bohai Bay and then characterized the enzymatic and catalytic properties of the enzyme. Finally, recombinant extracellular expression of this gene was carried out in Bacillus subtilis. The results showed that a novel fructan sucrase, BhLS 39, was mined from Bacillus halotolerans. With sucrose and raffinose as substrates, BhLS 39 showed the optimal temperatures of 50 ℃ and 55 ℃, optimal pH 5.5 for both, and Kcat/Km ratio of 3.4 and 6.6 L/(mmol·s), respectively. When 400 g/L raffinose was used as the substrate, the melibiose conversion rate was 84.6% after 30 min treatment with 5 U BhLS 39. Furthermore, BhLS 39 catalyzed the conversion of sucrose to produce levan-type-fructooligosaccharide and levan. Then, the recombinant extracellular expression of BhLS 39 in B. subtilis was achieved. The co-expression of the intracellular chaperone DnaK and the extracellular chaperone PrsA increased the extracellular activity of the recombinant BhLS 39 by 5.2 folds to 17 U/mL compared with that of the control strain. BhLS 39 obtained in this study is conducive to improving the quality and economic benefits of soybean oligosaccharides. At the same time, the strategy used here to enhance the extracellular expression of BhLS 39 will also promote the efficient recombinant expression of other proteins in B. subtilis.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"333-351"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041877","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Directed evolution improves the catalytic activity of laccase in papermaking].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240387

Hong Ni, Fan Yang, Lei Wang, Bianxia Li, Huanan Li, Jiashu Liu, Zhengbing Jiang, Wanli Cheng

{"title":"[Directed evolution improves the catalytic activity of laccase in papermaking].","authors":"Hong Ni, Fan Yang, Lei Wang, Bianxia Li, Huanan Li, Jiashu Liu, Zhengbing Jiang, Wanli Cheng","doi":"10.13345/j.cjb.240387","DOIUrl":"https://doi.org/10.13345/j.cjb.240387","url":null,"abstract":"As a biocatalyst, laccase has been widely studied and applied in the papermaking industry. However, the low catalytic efficiency and poor stability of natural laccase limit its application in the pulping process. To develop the laccase with high activity and strong tolerance, we carried out directed evolution for modification of the laccase derived from Bacillus pumilus and screened out the mutants F282L/F306L and Q275P from the random mutant library by high-throughput screening. The specific activities of F282L/F306L and Q275P were 280.87 U/mg and 453.94 U/mg, respectively, which were 1.42 times and 2.30 times that of the wild-type laccase. Q275P demonstrated significantly improved thermal stability, with the relative activity 20% higher than that of the wild-type laccase after incubation at 40 ℃, 50 ℃, and 70 ℃ for 4 h. F282L/F306L and Q275P showed greater tolerance to metal ions and organic solvents than the wild-type laccase. The Km value of the wild-type laccase was 374.97 μmo/L, and those of F282L/F306L and Q275P were reduced to 318.96 μmo/L and 360.71 μmo/L, respectively, which suggested that the substrate affinity of laccase was improved after mutation. The kcat values of F282L/F306L and Q275P for the substrate ABTS were 574.00 s-1 and 898.03 s-1, respectively, which were 1.1 times and 1.7 times that of the wild-type laccase, indicating the improved catalytic efficiency. Q275P demonstrated better performance than the wild-type laccase in pulping, as manifested by the reduction of 0.82 in the Kappa number and the increases of 2.00% ISO, 7.8%, and 7.2% in whiteness, tensile index, and breaking length, respectively. This work lays a foundation for improving the adaptation of laccase to the environment of the papermaking industry.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"308-320"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Metabolic engineering of Escherichia coli for efficient biosynthesis of L-citrulline].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240190

Linfeng Xu, Wenwen Yu, Xuewen Zhu, Quanwei Zhang, Yaokang Wu, Jianghua Li, Guocheng DU, Xueqin Lv, Jian Chen, Long Liu

{"title":"[Metabolic engineering of Escherichia coli for efficient biosynthesis of L-citrulline].","authors":"Linfeng Xu, Wenwen Yu, Xuewen Zhu, Quanwei Zhang, Yaokang Wu, Jianghua Li, Guocheng DU, Xueqin Lv, Jian Chen, Long Liu","doi":"10.13345/j.cjb.240190","DOIUrl":"https://doi.org/10.13345/j.cjb.240190","url":null,"abstract":"L-citrulline is a nonprotein amino acid that plays an important role in human health and has great market demand. Although microbial cell factories have been widely used for biosynthesis, there are still challenges such as genetic instability and low efficiency in the biosynthesis of L-citrulline. In this study, an efficient, plasmid-free, non-inducible L-citrulline-producing strain of Escherichia coli BL21(DE3) was engineered by combined strategies. Firstly, a chassis strain capable of synthesizing L-citrulline was constructed by block of L-citrulline degradation and removal of feedback inhibition, with the L-citrulline titer of 0.43 g/L. Secondly, a push-pull-restrain strategy was employed to enhance the L-citrulline biosynthesis, which realized the L-citrulline titer of 6.0 g/L. Thirdly, the NADPH synthesis and L-citrulline transport were strengthened to promote the synthesis efficiency, which achieved the L-citrulline titer of 11.6 g/L. Finally, fed-batch fermentation was performed with the engineered strain in a 3 L fermenter, in which the L-citrulline titer reached 44.9 g/L. This study lays the foundation for the industrial production of L-citrulline and provides insights for the modification of other amino acid metabolic networks.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"242-255"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Metabolic engineering of Escherichia coli for the biosynthesis of O-acetyl-L-homoserine].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240205

Lianggang Huang, Feng Gao, Nuoran Xu, Junping Zhou, Kun Niu, Bo Zhang, Zhiqiang Liu, Yuguo Zheng

{"title":"[Metabolic engineering of Escherichia coli for the biosynthesis of O-acetyl-L-homoserine].","authors":"Lianggang Huang, Feng Gao, Nuoran Xu, Junping Zhou, Kun Niu, Bo Zhang, Zhiqiang Liu, Yuguo Zheng","doi":"10.13345/j.cjb.240205","DOIUrl":"https://doi.org/10.13345/j.cjb.240205","url":null,"abstract":"O-acetyl-L-homoserine (OAH) is a promising platform compound for the production of L-methionine and other valuable compounds, while its low yield and low conversion rate limit the industrial application. To solve these problems, we constructed a strain for high OAH production with the previously constructed L-homoserine producer Escherichia coli HS33 as the chassis by systematic metabolic engineering. Firstly, PEP accumulation, pyruvate utilization, and OAH synthesis pathway (overexpressing aspB, aspA, and thrAC1034T) were enhanced to obtain an initial strain accumulating 13.37 g/L OAH. Subsequently, the co-factor synthesis genes were integrated to supply reducing power and energy, which increased the yield to 15.79 g/L. The OAH yield of the engineered strain OAH28 was further increased to 17.49 g/L by strengthening the acetic acid reuse pathway, improving the supply of acetyl-CoA, and regulating the expression of MetX from different sources. Finally, in a 5 L fermenter, OAH28 achieved an OAH titer of 47.12 g/L, with a glucose conversion rate of 32% and productivity of 0.59 g/(L·h). The results lay a foundation for increasing the OAH production by metabolic engineering and give insights into the industrial production of OAH.","PeriodicalId":21778,"journal":{"name":"Sheng wu gong cheng xue bao = Chinese journal of biotechnology","volume":"41 1","pages":"256-270"},"PeriodicalIF":0.0,"publicationDate":"2025-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143041871","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

[Research progress in tolerance of petroleum hydrocarbon pollutant-degrading strains].

Sheng wu gong cheng xue bao = Chinese journal of biotechnology Pub Date : 2025-01-25 DOI: 10.13345/j.cjb.240350

Shanshan Wang, Xiaoqian Zhu, Zhibei Cao, Lu Wang, Mingzhu Ding

引用次数: 0