Grosse-Honebrink Alexander, Little Gareth T, Bean Zak, Heldt Dana, Cornock Ruth H M, Winzer Klaus, Minton Nigel P, Green Edward, Zhang Ying
Clostridia Research Group, BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), Biodiscovery Institute, School of Life Sciences, University of Nottingham, Nottingham, United Kingdom.
CHAIN Biotechnology Ltd., MediCity, Nottingham, United Kingdom.
Front Bioeng Biotechnol. 2021 May 13;9:659895. doi: 10.3389/fbioe.2021.659895. eCollection 2021.
Chirally pure ()-1,3-butanediol (()-1,3-BDO) is a valuable intermediate for the production of fragrances, pheromones, insecticides and antibiotics. Biotechnological production results in superior enantiomeric excess over chemical production and is therefore the preferred production route. In this study ()-1,3-BDO was produced in the industrially important whole cell biocatalyst through expression of the enantio-specific gene from . The heterologous pathway was optimised in three ways: at the transcriptional level choosing strongly expressed promoters and comparing plasmid borne with chromosomal gene expression, at the translational level by optimising the codon usage of the gene to fit the inherent codon adaptation index of , and at the enzyme level by introducing point mutations which led to increased enzymatic activity. The resulting whole cell catalyst produced up to 20 mM (1.8 g/l) ()-1,3-BDO in non-optimised batch fermentation which is a promising starting position for economical production of this chiral chemical.
手性纯的()-1,3-丁二醇(()-1,3-BDO)是生产香料、信息素、杀虫剂和抗生素的重要中间体。生物技术生产比化学合成能产生更高的对映体过量,因此是首选的生产路线。在本研究中,通过表达来自的对映体特异性基因,在具有工业重要性的全细胞生物催化剂中生产()-1,3-BDO。异源途径通过三种方式进行优化:在转录水平,选择强表达启动子并比较质粒携带基因与染色体基因的表达;在翻译水平,通过优化基因的密码子使用以适应的固有密码子适应指数;在酶水平,通过引入导致酶活性增加的点突变。在非优化的分批发酵中,所得全细胞催化剂可产生高达20 mM(1.8 g/l)的()-1,3-BDO,这是该手性化学品经济生产的一个有前景的起始点。