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利用代谢工程改造的植物乳杆菌从糙米中生产光学纯D-乳酸。

Production of optically pure D-lactic acid from brown rice using metabolically engineered Lactobacillus plantarum.

作者信息

Okano Kenji, Hama Shinji, Kihara Maki, Noda Hideo, Tanaka Tsutomu, Kondo Akihiko

机构信息

Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka, 565-0871, Japan.

Bio-energy Corporation, Research & Development Laboratory, 2-9-7 Minaminanamatsu, Amagasaki, 660-0053, Japan.

出版信息

Appl Microbiol Biotechnol. 2017 Mar;101(5):1869-1875. doi: 10.1007/s00253-016-7976-8. Epub 2016 Nov 10.

Abstract

Simultaneous saccharification and fermentation (SSF) of D-lactic acid was performed using brown rice as both a substrate and a nutrient source. An engineered Lactobacillus plantarum NCIMB 8826 strain, in which the ʟ-lactate dehydrogenase gene was disrupted, produced 97.7 g/L D-lactic acid from 20% (w/v) brown rice without any nutrient supplementation. However, a significant amount of glucose remained unconsumed and the yield of lactic acid was as low as 0.75 (g/g-glucose contained in brown rice). Interestingly, the glucose consumption was significantly improved by adapting L. plantarum cells to the low-pH condition during the early stage of SSF (8-17 h). As a result, 117.1 g/L D-lactic acid was produced with a high yield of 0.93 and an optical purity of 99.6% after 144 h of fermentation. SSF experiments were repeatedly performed for ten times and D-lactic acid was stably produced using recycled cells (118.4-129.8 g/L). On average, D-lactic acid was produced with a volumetric productivity of 2.18 g/L/h over 48 h.

摘要

以糙米作为底物和营养源进行了D-乳酸的同步糖化发酵(SSF)。一株工程化的植物乳杆菌NCIMB 8826菌株,其L-乳酸脱氢酶基因被破坏,在不添加任何营养物质的情况下,从20%(w/v)的糙米中产生了97.7 g/L的D-乳酸。然而,大量葡萄糖仍未被消耗,乳酸产量低至0.75(g/g-糙米中所含葡萄糖)。有趣的是,在SSF早期阶段(8-17小时)使植物乳杆菌细胞适应低pH条件可显著改善葡萄糖消耗。结果,发酵144小时后产生了117.1 g/L的D-乳酸,产率高达到0.93,光学纯度为99.6%。SSF实验重复进行了十次,使用回收细胞稳定产生了D-乳酸(118.4-129.8 g/L)。平均而言,在48小时内D-乳酸的体积产率为2.18 g/L/h。

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