通过多级代谢工程提高淀粉样芽孢杆菌合成 S-腺苷蛋氨酸的能力。

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Letters Pub Date : 2024-08-20 DOI:10.1007/s10529-024-03523-x

Cong Jiang, Dian Zou, Liying Ruan, Wenyuan Han, Xuetuan Wei

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摘要

目的：结果：在淀粉芽孢杆菌 HSAM6 中，缺失 argG 基因阻止天冬氨酸分支降解，使 SAM 滴度增加到 254.78 ± 15.15：结果：在淀粉芽孢杆菌 HSAM6 中，删除阻断天冬氨酸分支降解的 argG 基因可将 SAM 滴度提高到 254.78 ± 15.91 mg/L，比 HSAM6 提高了 18%。随后，删除 moaA 基因以增加 5-甲基四氢叶酸的供应，导致生长受阻，SAM 产量急剧下降。通过过表达 citA 基因，菌株的生长得到了进一步改善，但 SAM 的合成并没有显著提高。最后，使用多拷贝质粒过表达 SAM2 基因获得了最高的 SAM 滴度（452.89 ± 13.42 mg/L）：结论：删除 argG 基因和过表达 SAM2 基因可显著提高淀粉芽孢杆菌的 SAM 合成能力。

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Multilevel metabolic engineering for enhanced synthesis of S-adenosylmethionine by Bacillus amyloliquefaciens.

Objectives: To enhance the de novo synthesis of SAM, the effects of several key genes on SAM synthesis were examined based on modular strategy, and the key genes were manipulated to obtain an engineered strain with high SAM production.

Results: In Bacillus amyloliquefaciens HSAM6, the deletion of argG gene to block aspartic acid branching degradation increased SAM titer to 254.78 ± 15.91 mg/L, up 18% from HSAM6. Subsequently, deleting the moaA gene to boost the supply of 5-methyltetrahydrofolate led to the stunted growth and the plummeting yield of SAM. Further improvement of strain growth by overexpression of the citA gene, while SAM synthesis was not significantly enhanced. Finally, the maximum SAM titer (452.89 ± 13.42 mg/L) was obtained by overexpression SAM2 gene using the multicopy plasmid.

Conclusions: The deletion of argG gene and the overexpression of SAM2 gene significantly improved SAM synthesis in B. amyloliquefaciens.

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.