在枯草芽孢杆菌中生产 2,3-丁二醇的不同脱氢酶的硅学筛选和验证

IF 2.5 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology and Bioprocess Engineering Pub Date : 2024-03-13 DOI:10.1007/s12257-024-00053-1
Sailee Sanjay Asolkar, M. Anju, Ravindra Kumar, Apoorva Deshmukh, Anand Ghosalkar, Pramod Kumbhar
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引用次数: 0

摘要

枯草芽孢杆菌是 2,3-丁二醇(2,3-BDO)的天然生产者,已获得 "公认安全 "地位。据报道,它能从合成糖以及糖蜜等复杂而经济的糖源中生产 2,3-BDO。不过,2,3-BDO 生成的限速步骤是由丁二醇脱氢酶(2,3-BDH)将乙炔醛转化为 2,3-BDO。筛选这类 2,3-BDH 的依据是它们对作为底物的乙炔酐具有较高的亲和力(较低的 Km)。为进一步验证,进行了硅对接研究,结果表明 PpBDH 蛋白与乙炔有较高的相互作用。在工程枯草芽孢杆菌(BS1A1)中对这些基因的异源表达进行了研究。据报道,在这项研究中,来自多粘芽孢杆菌 ZJ-9 的 2,3-BDH 具有更高的酶活性水平,而且在发酵研究中,2,3-BDO 与乙炔的比例提高了 80.25%。这些见解鼓励人们进一步优化生物工艺,以提高 2,3-BDO的发酵产量。我们的研究结果为避免在工程芽孢杆菌系统中将 2,3-BDO反向转化为乙酰丙酮提供了一种潜在的策略。
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In silico screening and validation of different dehydrogenases to produce 2,3-butanediol in Bacillus subtilis

Bacillus subtilis is a natural producer of 2,3-butanediol (2,3-BDO) and has acquired “Generally Regarded as Safe" status. It is reported to produce 2,3-BDO from synthetic sugars as well as complex and economic sugar sources such as molasses. However, the rate-limiting step in the formation of 2,3-BDO is its conversion from acetoin to 2,3-BDO by the enzyme butanediol dehydrogenase (2,3-BDH). Such 2,3-BDHs were screened based on higher affinity (lower Km) towards acetoin as substrate. The in silico docking studies were conducted for further validation, and they showed a high interaction profile for the PpBDH protein towards acetoin. Heterologous expression of these genes was studied in engineered Bacillus subtilis (BS1A1). In this study, it was seen that 2,3-BDH from Paenibacillus polymyxa ZJ-9 was reported to have higher enzyme activity levels, and in the fermentation studies, it was seen that the ratio of 2,3-BDO to acetoin was increased by 80.25%. The insights encourage further bioprocess optimization for increasing the fermentative production of 2,3-BDO. Our results provide a potential strategy to avoid the back conversion of 2,3-BDO to acetoin in an engineered Bacillus system.

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来源期刊
Biotechnology and Bioprocess Engineering
Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物
CiteScore
5.00
自引率
12.50%
发文量
79
审稿时长
3 months
期刊介绍: Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.
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