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类LsrR蛋白通过调节植物乳杆菌中的多糖生物合成来响应胁迫耐受性。

LsrR-like protein responds to stress tolerance by regulating polysaccharide biosynthesis in Lactiplantibacillus plantarum.

作者信息

Meng Fanqiang, Lyu Yunbin, Zhao Hongyuan, Lyu Fengxia, Bie Xiaomei, Lu Yingjian, Zhao Mingwen, Chen Yihua, Lu Zhaoxin

机构信息

College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing 210095, China; Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, Microbiology Department, College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China.

College of Food Science and Technology, Nanjing Agricultural University, 1 Weigang Road, Nanjing 210095, China.

出版信息

Int J Biol Macromol. 2023 Jan 15;225:1193-1203. doi: 10.1016/j.ijbiomac.2022.11.180. Epub 2022 Nov 25.

Abstract

In addition to their biological functions, polysaccharides assist Lactiplantibacillus plantarum in resisting harsh conditions. To enhance the polysaccharide biosynthesis and increase the survival of L. plantarum in gut environment. We analyzed the transcriptional regulators that regulated the polysaccharide biosynthesis. A new transcriptional inhibitor, LsrR (UniProtKB: Q88YH7), had been identified, which repressed polysaccharide synthesis by binding to the polysaccharide synthesis promoter cps4A-J (P). The EPSs and CPSs production of L. plantarum 163 was reduced by 42 % and 36 % (p < 0.05), respectively, when lsrR was overexpressed. Furthermore, alkaline shock proteins Asp2 and Asp1, heat shock protein Hsp3, and an autoinducer-2 (AI-2) related quorum-sensing regulator Rrp6 recovered the synthesis of polysaccharides to 50, 33, 55, and 60 %, respectively, by inhibiting the LsrR activity. This suggested that LsrR regulates polysaccharide synthesis in response to external stress signals such as pH, temperature, and AI-2 concentration. Finally, we showed that polysaccharides increased the survival rate of L. plantarum (Lp163-ΔlsrR) by 2.1 times during lyophilization and enhanced its tolerance to pH 2.0 and 0.2 % bile salts by 15.3 and 60 times due to increased capsular thickness and enhanced the autoaggregation. We provide critical data regarding Lactobacillus survival during preservative lyophilization and under gastrointestinal conditions.

摘要

除了其生物学功能外,多糖还能帮助植物乳杆菌抵抗恶劣条件。为了增强多糖生物合成并提高植物乳杆菌在肠道环境中的存活率,我们分析了调控多糖生物合成的转录调节因子。已鉴定出一种新的转录抑制剂LsrR(UniProtKB:Q88YH7),它通过与多糖合成启动子cps4A-J(P)结合来抑制多糖合成。当lsrR过表达时,植物乳杆菌163的胞外多糖(EPSs)和荚膜多糖(CPSs)产量分别降低了42%和36%(p<0.05)。此外,碱性休克蛋白Asp2和Asp1、热休克蛋白Hsp3以及一种与自诱导物-2(AI-2)相关的群体感应调节因子Rrp6通过抑制LsrR活性,分别将多糖合成恢复到50%、33%、55%和60%。这表明LsrR响应pH、温度和AI-2浓度等外部应激信号来调节多糖合成。最后,我们发现多糖在冻干过程中将植物乳杆菌(Lp163-ΔlsrR)的存活率提高了2.1倍,并且由于荚膜厚度增加和自聚集增强,使其对pH 2.0和0.2%胆盐的耐受性分别提高了15.3倍和60倍。我们提供了关于乳酸菌在冻干保存和胃肠道条件下存活的关键数据。

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