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固定化植物乳杆菌谷氨酸脱羧酶在米醋和味精体系中生产食品级γ-氨基丁酸。

Food-grade γ-aminobutyric acid production by immobilized glutamate decarboxylase from Lactobacillus plantarum in rice vinegar and monosodium glutamate system.

机构信息

College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, 163319, China.

School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou, 310023, China.

出版信息

Biotechnol Lett. 2021 Oct;43(10):2027-2034. doi: 10.1007/s10529-021-03164-4. Epub 2021 Jul 26.

Abstract

OBJECTIVES

γ-amino butyric acid (GABA) is a non-protein amino acid, considered a potent bioactive compound. This study focused on biosynthesis of food-grade GABA by immobilized glutamate decarboxylase (GAD) from Lactobacillus plantarum in the rice vinegar and monosodium glutamate (MSG) reaction system.

RESULTS

The gene encoding glutamate decarboxylase (GadB) from L. plantarum has been heterologously expressed in Lactococcus lactis and biochemically characterized. Recombinant GadB existed as a homodimer, and displayed maximal activity at 40 °C and pH 5.0. The K value and catalytic efficiency (k/K) of GadB for L-Glu was 22.33 mM and 62.4 mM min, respectively, with a specific activity of 24.97 U/mg protein. Then, purified GadB was encapsulated in gellan gum beads. Compared to the free enzyme, immobilized GadB showed higher operational and storage stability. Finally, 9.82 to 21.48 g/L of GABA have been acquired by regulating the amounts of catalyst microspheres ranging from 0.5 to 0.8 g (wet weight) in 0.8 mL of the designed rice vinegar and MSG reaction system.

CONCLUSIONS

The method of production GABA by immobilized GadB microspheres mixed in the rice vinegar and MSG reaction system is introduced herein for the first time. Especially, the results obtained here meet the increased interest in the harnessing of biocatalyst to synthesize food-grade GABA.

摘要

目的

γ-氨基丁酸(GABA)是一种非蛋白质氨基酸,被认为是一种有效的生物活性化合物。本研究专注于通过固定化谷氨酸脱羧酶(GAD)从植物乳杆菌在米醋和味精(MSG)反应体系中生产食品级 GABA。

结果

已在乳球菌乳球菌中异源表达了来自植物乳杆菌的谷氨酸脱羧酶(GadB)基因,并进行了生化表征。重组 GadB 作为同源二聚体存在,在 40°C 和 pH 5.0 时显示最大活性。GadB 对 L-Glu 的 K 值和催化效率(k/K)分别为 22.33 mM 和 62.4 mM·min,比活为 24.97 U/mg 蛋白。然后,将纯化的 GadB 包埋在结冷胶珠中。与游离酶相比,固定化 GadB 显示出更高的操作稳定性和储存稳定性。最后,通过调节催化剂微球的量(湿重 0.5 至 0.8 g),在设计的米醋和 MSG 反应体系中,可获得 9.82 至 21.48 g/L 的 GABA。

结论

本文首次介绍了通过固定化 GadB 微球在米醋和 MSG 反应体系中混合生产 GABA 的方法。特别是,这里获得的结果满足了人们对利用生物催化剂合成食品级 GABA 的日益增长的兴趣。

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