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多粘芽孢杆菌中吲哚-3-乙酸合成途径的鉴定与组合工程

Identification and combinatorial engineering of indole-3-acetic acid synthetic pathways in Paenibacillus polymyxa.

作者信息

Sun Huimin, Zhang Jikun, Liu Wenteng, E Wenhui, Wang Xin, Li Hui, Cui Yanru, Zhao Dongying, Liu Kai, Du Binghai, Ding Yanqin, Wang Chengqiang

机构信息

College of Life Sciences, Shandong Key Laboratory of Agricultural Microbiology, State Key Laboratory of Crop Biology, Shandong Engineering Research Center of Plant-Microbia Restoration for Saline-Alkali Land, Shandong Agricultural University, 61 Daizong Street, Tai'an, 271018, China.

出版信息

Biotechnol Biofuels Bioprod. 2022 Aug 11;15(1):81. doi: 10.1186/s13068-022-02181-3.

Abstract

BACKGROUND

Paenibacillus polymyxa is a typical plant growth-promoting rhizobacterium (PGPR), and synthesis of indole-3-acetic acid (IAA) is one of the reasons for its growth-promoting capacity. The synthetic pathways of IAA in P. polymyxa must be identified and modified.

RESULTS

P. polymyxa SC2 and its spontaneous mutant SC2-M1 could promote plant growth by directly secreting IAA. Through metabonomic and genomic analysis, the genes patA, ilvB3, and fusE in the native IPyA pathway of IAA synthesis in strain SC2-M1 were predicted. A novel strong promoter P was rationally selected, synthetically analyzed, and then evaluated on its ability to express IAA synthetic genes. Co-expression of three genes, patA, ilvB3, and fusE, increased IAA yield by 60% in strain SC2-M1. Furthermore, the heterogeneous gene iaam of the IAM pathway and two heterogeneous IPyA pathways of IAA synthesis were selected to improve the IAA yield of strain SC2-M1. The genes ELJP6_14505, ipdC, and ELJP6_00725 of the entire IPyA pathway from Enterobacter ludwigii JP6 were expressed well by promoter P in strain SC2-M1 and increased IAA yield in the engineered strain SC2-M1 from 13 to 31 μg/mL, which was an increase of 138%.

CONCLUSIONS

The results of our study help reveal and enhance the IAA synthesis pathways of P. polymyxa and its future application.

摘要

背景

多粘芽孢杆菌是一种典型的促进植物生长的根际细菌(PGPR),吲哚 - 3 - 乙酸(IAA)的合成是其具有促生长能力的原因之一。必须确定并改造多粘芽孢杆菌中IAA的合成途径。

结果

多粘芽孢杆菌SC2及其自发突变体SC2 - M1可通过直接分泌IAA促进植物生长。通过代谢组学和基因组分析,预测了菌株SC2 - M1中IAA合成的天然IPyA途径中的patA、ilvB3和fusE基因。合理选择了一个新型强启动子P,进行了综合分析,然后评估其表达IAA合成基因的能力。patA、ilvB3和fusE这三个基因共表达使菌株SC2 - M1中的IAA产量提高了60%。此外,选择了IAA合成的IAM途径的异源基因iaam和两条异源IPyA途径来提高菌株SC2 - M1的IAA产量。来自路德维希肠杆菌JP6的整个IPyA途径的基因ELJP6_14505、ipdC和ELJP6_00725在菌株SC2 - M1中通过启动子P良好表达,并使工程菌株SC2 - M1中的IAA产量从13μg/mL提高到31μg/mL,提高了138%。

结论

我们的研究结果有助于揭示和增强多粘芽孢杆菌的IAA合成途径及其未来应用。

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