通过蛋白质组学分析揭示甜菜碱对脱氮假单胞菌维生素 B 生物合成途径的促进作用。

Revealing the Promoting Effect of Betaine on Vitamin B Biosynthetic Pathway of Pseudomonas denitrificans by Using a Proteomics Analysis.

机构信息

College of Food Science and Technology, Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety, Guangdong Ocean University, Zhanjiang, 524088, China | Jiangxi Engineering Laboratory for the Development and Utilization of Agricultural Microbial Resources, Jiangxi Agricultural University, Nanchang, 330045, China.

University of Shanghai for Science and Technology, Shanghai Engineering Research Center of Food Microbiology, Shanghai, 200093, China.

出版信息

Curr Pharm Biotechnol. 2022;23(3):466-475. doi: 10.2174/1389201022666210531120935.

DOI:10.2174/1389201022666210531120935

PMID:34061014

Abstract

BACKGROUND

Our previous comparative metabolomics research revealed that betaine (N,N,Ntrimethylglycine, a typically essential methyl-group donor for vitamin B biosynthesis) had powerful promoting effect on the generation of vitamin B precursors and intermediates in vitamin B-producing Pseudomonas denitrificans. However, the integral effect of betaine on the vitamin B12 biosynthetic pathway is still unclear.

OBJECTIVES

Considering the vitamin B biosynthetic pathway of P. denitrificans as a whole, this work aimed to reveal the biological function of betaine on the vitamin B biosynthetic pathway in P. denitrificans, which would sharpen and expand understanding of betaine as the methyl-group donor for vitamin B biosynthesis.

MATERIALS AND METHODS

By using a proteomics method based on the iTRAQ technique, the present study compared and analyzed the differential expression of proteins involved in vitamin B biosynthetic pathway under 10 g/L betaine in addition to P. denitrificans fermentation medium.

RESULTS

The results showed that betaine could significantly up-regulate the expression of proteins related to the vitamin B biosynthetic pathway, which was mainly reflected in the following three aspects: 1) the δ-aminolevulinic acid (ALA) synthase and porphobilinogen synthase that were responsible for the formation of the committed precursors for tetrapyrrole-derived macrocycle in vitamin B12 molecule; 2) the C-methylation-related enzymes (such as precorrin-4 C(11)-methyltransferase, precorrin-2 C(20)- methyltransferase, precorrin-8X methylmutase, and precorrin-6Y C5,15-methyltransferase) and methionine synthase that were crucial to the C-methylation reactions for vitamin B biosynthesis; 3) the latestage key enzymes (Cobaltochelatase, and Cob(I)yrinic acid a,c-diamide adenosyltransferase) that were related to cobalt chelation of vitamin B molecule.

CONCLUSION

The present study demonstrated clearly that betaine could significantly promote the expression of the integral enzymes involved in the vitamin B biosynthetic pathway of P. denitrificans, thus promoting vitamin B biosynthesis.

摘要

背景

我们之前的比较代谢组学研究表明，甜菜碱（N，N，N-三甲基甘氨酸，维生素 B 生物合成的典型必需甲基供体）对产维生素 B 的假单胞菌中维生素 B 前体和中间体的生成具有强大的促进作用。然而，甜菜碱对维生素 B12 生物合成途径的整体影响尚不清楚。

目的

考虑到假单胞菌的维生素 B 生物合成途径是一个整体，本工作旨在揭示甜菜碱对假单胞菌维生素 B 生物合成途径的生物学功能，这将深化和扩展对甜菜碱作为维生素 B 生物合成甲基供体的认识。

材料和方法

通过基于 iTRAQ 技术的蛋白质组学方法，本研究比较和分析了在 10 g/L 甜菜碱和假单胞菌发酵培养基之外，参与维生素 B 生物合成途径的蛋白质的差异表达。

结果

结果表明，甜菜碱可以显著上调与维生素 B 生物合成途径相关的蛋白质的表达，这主要体现在以下三个方面：1）δ-氨基乙酰丙酸（ALA）合酶和卟胆原合酶，负责四吡咯衍生大环中维生素 B12 分子的形成的关键前体；2）C-甲基化相关酶（如前原啉-4 C（11）-甲基转移酶、前原啉-2 C（20）-甲基转移酶、前原啉-8X 甲基突变酶和前原啉-6Y C5，15-甲基转移酶）和甲硫氨酸合酶，这些酶对维生素 B 生物合成的 C-甲基化反应至关重要；3）后期关键酶（钴酰基辅酶酶和钴（I）yrinic 酸 a，c-二酰胺腺苷转移酶），与维生素 B 分子的钴螯合有关。