Metabolic engineering for the production of acetoin and 2,3-butanediol at elevated temperature in Parageobacillus thermoglucosidasius NCIMB 11955.
Metabolic engineering for the production of acetoin and 2,3-butanediol at elevated temperature in Parageobacillus thermoglucosidasius NCIMB 11955.
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热糖sidasiparageobacillus NCIMB 11955在高温下生产乙酰和2,3-丁二醇的代谢工程。
DOI：
10.3389/fbioe.2023.1191079
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发表时间：
2023
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
--
作者：
当前的气候危机强调了到2050年实现全球净零排放的必要性，并敦促各国到2030年制定大量减排目标。利用使用嗜热底盘的发酵过程可以代表一种通过更环保的路线制造化学品和燃料的方式，同时净减少温室气体排放。在这项研究中，工业上相关的嗜热副芽孢杆菌热葡糖苷酶NCIMB 11955被工程化以生产3-羟基丁酮（乙偶姻）和2，3-丁二醇（2，3-BDO），这是具有商业应用的有机化合物。使用异源乙酰乳酸合酶（ALS）和乙酰乳酸脱羧酶（ALD），构建功能性2，3-BDO生物合成途径。通过删除丙酮酸节点周围的竞争途径，使副产物的形成最小化。氧化还原不平衡通过自主过表达的丁二醇脱氢酶，并通过调查适当的通风水平。通过这一点，我们能够产生2，3-BDO作为主要的发酵代谢物，其中高达6.6 g/L 2，3-BDO（0.33 g/g葡萄糖）代表50°C下理论最大值的66%。此外，以前未报道的嗜热乙偶姻降解基因（acoB 1）的鉴定和随后的删除导致在有氧条件下乙偶姻生产增强，产生7.6 g/L（0.38 g/g葡萄糖），代表理论最大值的78%。此外，通过产生Δ acoB 1突变体并通过测试葡萄糖浓度对2，3-BDO产生的影响，我们能够在补充有5%葡萄糖的培养基中产生15.6 g/L的2，3-BDO，这是迄今为止副芽孢杆菌和土芽孢杆菌属物种中产生的2，3-BDO的最高滴度。
The current climate crisis has emphasised the need to achieve global net-zero by 2050, with countries being urged to set considerable emission reduction targets by 2030. Exploitation of a fermentative process that uses a thermophilic chassis can represent a way to manufacture chemicals and fuels through more environmentally friendly routes with a net reduction in greenhouse gas emissions. In this study, the industrially relevant thermophile Parageobacillus thermoglucosidasius NCIMB 11955 was engineered to produce 3-hydroxybutanone (acetoin) and 2,3-butanediol (2,3-BDO), organic compounds with commercial applications. Using heterologous acetolactate synthase (ALS) and acetolactate decarboxylase (ALD) enzymes, a functional 2,3-BDO biosynthetic pathway was constructed. The formation of by-products was minimized by the deletion of competing pathways surrounding the pyruvate node. Redox imbalance was addressed through autonomous overexpression of the butanediol dehydrogenase and by investigating appropriate aeration levels. Through this, we were able to produce 2,3-BDO as the predominant fermentation metabolite, with up to 6.6 g/L 2,3-BDO (0.33 g/g glucose) representing 66% of the theoretical maximum at 50°C. In addition, the identification and subsequent deletion of a previously unreported thermophilic acetoin degradation gene (acoB1) resulted in enhanced acetoin production under aerobic conditions, producing 7.6 g/L (0.38 g/g glucose) representing 78% of the theoretical maximum. Furthermore, through the generation of a ΔacoB1 mutant and by testing the effect of glucose concentration on 2,3-BDO production, we were able to produce 15.6 g/L of 2,3-BDO in media supplemented with 5% glucose, the highest titre of 2,3-BDO produced in Parageobacillus and Geobacillus species to date.
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影响因子：
6.3
作者：
Li C;Gai Z;Wang K;Jin L
通讯作者：
Jin L
影响因子：
8
作者：
Extance, Jonathan;Danson, Michael J.;Crennell, Susan J.
通讯作者：
Crennell, Susan J.
影响因子：
10.4
作者：
Allen JG;Flanigan SS;LeBlanc M;Vallarino J;MacNaughton P;Stewart JH;Christiani DC
通讯作者：
Christiani DC
影响因子：
3.3
作者：
Coleman, Gareth A.;Pancost, Richard D.;Williams, Tom A.
通讯作者：
Williams, Tom A.
DOI：
10.1107/s0907444913020349
发表时间：
2013-10-01
影响因子：
2.2
作者：
Extance, Jonathan;Crennell, Susan J.;Danson, Michael J.
通讯作者：
Danson, Michael J.