Bioinformatics Analysis and Gene Cloning of L-Lactate Dehydrogenase from Lactobacillus rhamnosus

被引：0

作者：

Xu L. ^{[1
]}

Jiang G. ^{[2
]}

Wu X. ^{[1
]}

Liao X. ^{[1
]}

Lao X. ^{[1
]}

Ya H. ^{[1
]}

Li X. ^{[1
]}

机构：

[1] College of Medicine and Health, Industrial Technology Engineering Center for Zhuang & Yao Medicinal Organisms, Botanical Garden of Zhuang & Yao Medicinal Plants, Colaborative Innovation Center of Great Health, Guangxi Vocational & Technical Institute of In

[2] College of Forestry, Guangxi Key Laboratory of Forest Ecology and Conservation, State Key Laboratory for Conservation, Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning

来源：

Science and Technology of Food Industry | 2023年 / 44卷 / 11期

关键词：

active site; bioinformatics; enzyme activity; evolutionary analysis; L-lactate dehydrogenase; Lactobacillus rhamnosus;

D O I：

10.13386/j.issn1002-0306.2022080109

中图分类号：

学科分类号：

摘要：

In this study, L-lactate dehydrogenase (Lr-L-LDH) from Lactobacillus rhamnosus was investigated. The well-known Lr-L-LDH1 was taken as the control, differences between Lr-L-LDH1 and Lr-L-LDH2 annotated from the genome of L. rhamnosus were analyzed. The primary structure, basic properties, hydrophobicity, secondary structure of Lr-L-LDH1 and Lr-L-LDH2 were predicted and analyzed using online websites and professional software. Homology modeling of the tertiary structure, molecular docking of enzymes and substrates, phylogenetic analysis, in vitro cloning, expression, and enzyme activity assays were further studied. The results showed that, in comparison to Lr-L-LDH1, although Lr-L-LHD2 had similar molecular characteristics, secondary and tertiary structures, Lr-L-LHD2 exhibited differences: A shorter sequences, low amino acid identities (48.08%), and a different phylogenetic status. Lr-L-LHD2 also contained a catalytically active site, a highly conserved NAD+ binding site sequence (GXGXXG), a three-dimensional active pocket domain, was an NAD+ dependent tetrameric L-lactate dehydrogenase, and be activated by fructose 1,6-diphosphate in the cytoplasm for catalyzing the reduction of pyruvate to L-lactic acid. In vitro enzyme activity was significantly lower than Lr-L-LDH1 (P<0.01). In conclusion, both Lr-L-LDH1 and Lr-L-LHD2 showed enzyme activities of L-lactate dehydrogenase, indicating they may regulate the L-lactate metabolism pathway together. Considerations of both Lr-L-LDH1 and Lr-LLHD2 should be taken for the future molecular modifications of L-LDH in L. rhamnosus, and the study provided the molecular and scientific basis for the gene engineering modifications of lactic acid fermentation industry. © 2023 Science and Technology of Food Industry. All rights reserved.

引用

页码：153 / 162

页数：9

共 41 条

[1]

JU S Y, KIM J H, LEE P C., Long-term adaptive evolution of Leuconostoc mesenteroides for enhancement of lactic acid tolerance and production[J], Biotechnology for Biofuels, 9, 1, (2016)

[2]

ABDEL-RAHMAN M A, TASHIRO Y, SONOMOTO K., Lactic acid production from lignocellulose-derived sugars using lactic acid bacteria: Overview and limits[J], Journal of Biotechnology, 156, 4, (2010)

[3]

PARRA-RAMIREZ D, MARTINEZ A, CARDONA C A., Lactic acid production from glucose and xylose using the lactogenic Escherichia coli strain JU15: Experiments and techno-economic results[J], Bioresource Technology, 273, (2018)

[4]

SUN H X, ZHOU W Q, YANG S, Et al., Progress in microbiological preparation of L-lactic acid strain with high optical purity and its production[J], Food and Fermentation Sciences & Technology, 57, 5, (2021)

[5]

LIU J X, LI G Y, JIN Q., Research progress in production of D-/L-lactic acid by lactic acid bacteria[J], China Food Safety Magazine, 33, (2021)

[6]

WESSELS S, AXELSSON L, HANSEN E B, Et al., The lactic acid bacteria, the food chain, and their regulation[J], Trends in Food Science & Technology, 15, 10, pp. 498-505, (2004)

[7]

BERNARD N, FERAIN T, GARMYN D, Et al., Cloning of the D-lactate dehydrogenase gene from Lactobacillus delbrueckii subsp. bulgaricus by complementation in Escherichia coli[J], FEBS Letters, 290, (1991)

[8]

ABDEL-RAHMAN M A, TASHIRO Y, SONOMOTO K., Recent advances in lactic acid production by microbial fermentation processes[J], Biotechnology Advances, 31, 6, (2013)

[9]

Recombinant strain producing L-lactic acid and its construction method, fermentation method and application

[10]

LI X Q, YUAN F J, LIU Y, Et al., Expression and bioinformatic analysis of an L-Lactate Dehydrogenase (L-LcLDH2) for the asymmetric reduction of phenylpyruvic acid[J], Journal of Food Science and Biotechnology, 38, 12, (2019)

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