A new l-serine binding orphan SerBP affects indole synthesis in Pantoea ananatis

文献类型： 外文期刊

第一作者： Zhang, Lei

作者： Zhang, Lei;Yu, Jiajia;Zheng, Jing;Zhou, Xinyi;Ban, Yali;Sun, Yuanhao;Zhang, Haotian;Feng, Yongjun;Wu, Liqing;Feng, Yongjun

作者机构：

关键词： amino acid transport; indole; Pantoea ananatis YJ76; substrate-binding protein

期刊名称：JOURNAL OF BASIC MICROBIOLOGY （ 影响因子：3.1； 五年影响因子：3.0 ）

ISSN： 0233-111X

年卷期： 2023 年

页码：

收录情况： SCI

摘要： Indole is traditionally known as a metabolite of l-tryptophan and now as an important signaling molecule in bacteria, however, the understanding of its upstream synthesis regulation is very limited. Pantoea ananatis YJ76, a predominant diazotrophic endophyte isolated from rice (Oryza sativa), can produce indole to regulate various physiological and biochemical behaviors. We constructed a mutant library of YJ76 using the mTn5 transposon insertion mutation method, from which an indole-deficient mutant was screened out. Via high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR), the transposon was determined to be inserted in a gene (RefSeq: WP014605468.1) of unknown function that is highly conserved at the intraspecific level. Bioinformatics analysis implied that the protein (Protein ID: WP089517194.1) encoded by the mutant gene is most likely to be a new orphan substrate-binding protein (SBP) for amino acid ABC transporters. Amino acid supplement cultivation experiments and surface plasmon resonance revealed that the protein could bind to l-serine (K-D = 6.149 x 10(-5) M). Therefore, the SBP was named as SerBP. This is the first case that a SBP responds to l-serine ABC transports. As a precursor of indole synthesis, the transmembrane transported l-serine was directly correlated with indole signal production and the mutation of serBP gene weakened the resistance of YJ76 to antibiotics, alkali, heavy metals, and starvation. This study provided a new paradigm for exploring the upstream regulatory pathway for indole synthesis of bacteria.

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