Ramakrishnan Gopi Gopal, Nehru Ganesh, Suppuram Pandiaraj, Balasubramaniyam Sowmiya, Gulab Brajesh Raman, Subramanian Ramalingam
Centre for Biotechnology, Anna University, Chennai, Tamil Nadu, 600 025, India.
Curr Microbiol. 2015 Oct;71(4):517-23. doi: 10.1007/s00284-015-0878-7. Epub 2015 Jul 24.
Lactobacillus reuteri grown in MRS broth containing 20 mM glycerol exhibits 3.7-fold up-regulation of 3-hydroxypropionic acid (3-HP) pathway genes during the stationary phase. Concomitantly, the resting cells prepared from stationary phase show enhancement in bio-conversion of glycerol, and the maximum specific productivity (q p) is found to be 0.17 g 3-HP per g CDW per hour. The regulatory elements such as catabolite repression site in the up-stream of 3-HP pathway genes are presumed for the augmentation of glycerol bio-conversion selectively in stationary phase. However, in the repression mutant, the maximum q p of 3-HP persisted in the stationary phase-derived resting cells indicating the role of further regulatory features. In the production stage, the external 3-HP concentration of 35 mM inhibits 3-HP synthesis. In addition, it has also moderated 1,3-propanediol formation, as it is a redox bio-catalysis involving NAD(+)/NADH ratio of 6.5. Repeated batch bio-transformation has been used to overcome product inhibition, and the total yield (Ypx) of 3-HP from the stationary phase-derived biomass is 3.3 times higher than that from the non-repeated mode. With the use of appropriate gene expression condition and repeated transfer of biomass, 3-HP produced in this study can be used for low-volume, high-value applications.
在含有20 mM甘油的MRS肉汤中生长的罗伊氏乳杆菌在稳定期3-羟基丙酸(3-HP)途径基因上调3.7倍。与此同时,从稳定期制备的静息细胞显示甘油生物转化增强,最大比生产率(qp)为每克细胞干重每小时0.17克3-HP。推测3-HP途径基因上游的分解代谢物阻遏位点等调控元件可在稳定期选择性增强甘油生物转化。然而,在阻遏突变体中,稳定期来源的静息细胞中3-HP的最大qp持续存在,表明存在进一步的调控特征。在生产阶段,35 mM的外部3-HP浓度会抑制3-HP合成。此外,它还减缓了1,3-丙二醇的形成,因为这是一种涉及NAD(+)/NADH比率为6.5的氧化还原生物催化过程。已采用重复批次生物转化来克服产物抑制,稳定期来源的生物质中3-HP的总产率(Ypx)比非重复模式高3.3倍。通过使用合适的基因表达条件和重复转移生物质,本研究中产生的3-HP可用于小批量、高价值应用。