Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, 565-0871, Osaka, Japan.
Division of Applied Science, School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, 565-0871 Osaka, Japan.
Biotechnol J. 2018 May;13(5):e1700517. doi: 10.1002/biot.201700517. Epub 2018 Feb 22.
Fermentative production of optically pure lactic acid (LA) has attracted great interest because of the increased demand for plant-based plastics. For cost-effective LA production, an engineered Lactobacillus plantarum NCIMB 8826 strain, which enables the production of optically pure l-LA from raw starch, is constructed. The wild-type strain produces a racemic mixture of d- and l-LA from pyruvate by the action of the respective lactate dehydrogenases (LDHs). Therefore, the gene encoding D-LDH (ldhD) is deleted. Although no decrease in d-LA formation is observed in the ΔldhD mutant, additional disruption of the operon encoding lactate racemase (larA-E), which catalyzes the interconversion between d- and l-LA, completely abolished d-LA production. From 100 g L glucose, the ΔldhD ΔlarA-E mutant produces 87.0 g L of l-LA with an optical purity of 99.4%. Subsequently, a plasmid is introduced into the ΔldhD ΔlarA-E mutant for the secretion of α-amylase from Streptococcus bovis 148. The resulting strain could produce 50.3 g L of l-LA from raw corn starch with a yield of 0.91 (g per g of consumed sugar) and an optical purity of 98.6%. The engineered L. plantarum strain would be useful in the production of l-LA from starchy materials.
由于对植物基塑料的需求增加,发酵生产光学纯乳酸(LA)引起了极大的兴趣。为了实现 LA 的经济高效生产,构建了一株能够从原始淀粉生产光学纯 l-LA 的工程化植物乳杆菌 NCIMB 8826 菌株。野生型菌株通过各自的乳酸脱氢酶(LDHs)作用,从丙酮酸生产 d-和 l-LA 的外消旋混合物。因此,编码 D-LDH(ldhD)的基因被删除。尽管在ΔldhD 突变体中没有观察到 d-LA 形成的减少,但进一步破坏编码乳酸 racemase(larA-E)的操纵子,该酶催化 d-和 l-LA 之间的互变,完全消除了 d-LA 的生产。从 100 g L 的葡萄糖中,ΔldhD ΔlarA-E 突变体产生 87.0 g L 的 l-LA,光学纯度为 99.4%。随后,将质粒引入ΔldhD ΔlarA-E 突变体中,用于从牛链球菌 148 分泌α-淀粉酶。得到的菌株可以从生玉米淀粉中生产 50.3 g L 的 l-LA,产率为 0.91(每消耗糖 g 产生的 g),光学纯度为 98.6%。该工程化的植物乳杆菌菌株可用于从淀粉质材料生产 l-LA。