New molecule of nonribosomal peptide synthesis mechanism from Lactiplantibacillus plantarum LMG100 probiotic bacteria

IF 0.7 Q4 PHARMACOLOGY & PHARMACY
Amr Elmasry, Walaa Hussein, Ali Abdelmoteleb
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引用次数: 0

Abstract

Background The huge increasing on gastrointestinal illness by spreading of resistance pathogens requires to develop alternative antimicrobial agents. Nonribosomal peptides are considered one of these alternatives which produced by wild spectrum of bacteria. Objective Detection of nonribosomal peptide synthesis from Lactiplantibacillus plantarum LMG100 probiotic strain isolated from traditional lactic fermenting foods as alternative antimicrobial agent is important to human health and immune system. Materials and methods The identification of isolated strains using 16S rDNA technique was performed and followed by bioinformatics analysis tools; AntiSmash, PKS-NRPS analysis website, LSI based A-domain function predictor, NRPS predictor2, clustering using PhyML 3.0 to detect adenylation domain substrate specificity of NRP synthetases genes cluster of Lactiplantibacillus plantarum LMG100. To prove the presence of the NRP synthetases genes cluster, degenerate primers protocol and three sets of primers covered the five gene cluster were designed based on the original reference strain L. plantarum WCFS1. Antibacterial activity of the isolated strain was detected against bacterial strains from coliform group of the enteric genera of Escherichia, Salmonella and Shigella which formed the ordinary gastrointestinal tract infection. Results and conclusion The isolated L. plantarum LMG100 strain showed 99.96% of identity to 16S rDNA partial gene sequence of Lactiplantibacillus plantarum WCFS1 strain and bioinformatics analysis tools revealed the presence of NRPS gene cluster of five genes; two biosynthetic genes npsA and npsB from the five genes encoded for polypeptide of six amino acids, but six different predictors programs couldn’t assign the specificity of all adenylation domains except A4 serine and A5 glycine. The use of degenerate primers confirmed the presence of the NRPS in the isolated strain L. plantarum LMG100 compared to the standard strain WCFS1. Three sets of primers covering the five gene cluster were designed based on the original reference strain L. plantarum WCFS1 complete genome sequence confirmed the same organization of the putative gene cluster. In general, the approach of degenerated primers proved the presence of polypeptide NRPs gene presence in lactobacilli isolates. The new polypeptide NRP produced from the strain LMG100 showed maximum inhibition zones against G-ve short-rods bacterial strains from the coliform group of the enteric genera of Escherichia, Salmonella and Shigella which formed the ordinary gastrointestinal tract infection, and the minimal inhibitory concentrations (MIC) for G-ve bacterial strains was approximately of 125 mg.ml−1. The obtained results revealed that the selected probiotic Lactiplantibacillus strain is suitable candidate for use as bio-preservative starter or probiotic for human consumption in food and pharmaceutical industries.
植物乳杆菌 LMG100 益生菌的非核糖体肽合成机制新分子
背景 由于抗药性病原体的传播,胃肠道疾病的发病率大幅上升,因此需要开发替代抗菌剂。由野生细菌产生的非核糖体肽被认为是这些替代品之一。从传统乳酸发酵食品中分离出的植物乳杆菌 LMG100 益生菌株合成非核糖体肽作为替代抗菌剂对人类健康和免疫系统非常重要。材料与方法 利用 16S rDNA 技术对分离菌株进行鉴定,然后利用生物信息学分析工具 AntiSmash、PKS-NRPS 分析网站、基于 LSI 的 A-domain 功能预测器、NRPS 预测器 2 和 PhyML 3.0 聚类分析工具检测植物乳杆菌 LMG100 的 NRP 合成酶基因簇的腺苷酸化域底物特异性。为了证明植物乳杆菌 LMG100 的 NRP 合成酶基因簇的存在,以原始参考菌株 L. plantarum WCFS1 为基础,设计了退化引物方案和三组涵盖五个基因簇的引物。检测了分离菌株对普通胃肠道感染的肠道埃希氏菌属、沙门氏菌属和志贺氏菌属大肠菌群细菌菌株的抗菌活性。结果与结论 分离出的植物乳杆菌 LMG100 菌株与植物乳杆菌 WCFS1 菌株的 16S rDNA 部分基因序列的同一性为 99.96%,生物信息学分析工具显示存在由五个基因组成的 NRPS 基因簇;五个基因中的两个生物合成基因 npsA 和 npsB 编码六个氨基酸的多肽,但六个不同的预测程序无法确定除 A4 丝氨酸和 A5 甘氨酸以外的所有腺苷酸化结构域的特异性。与标准菌株 WCFS1 相比,使用退化引物证实了分离菌株 L. plantarum LMG100 中 NRPS 的存在。根据原始参考菌株 L. plantarum WCFS1 的完整基因组序列,设计了涵盖五个基因簇的三组引物,证实了假定基因簇的组织结构相同。总的来说,退化引物的方法证明了乳酸菌分离物中存在多肽 NRPs 基因。从菌株 LMG100 中产生的新多肽 NRP 对形成普通胃肠道感染的肠道埃希氏菌属、沙门氏菌属和志贺氏菌属大肠菌群中的 G-ve 短杆细菌菌株显示出最大抑菌区,对 G-ve 细菌菌株的最小抑菌浓度(MIC)约为 125 毫克/毫升-1。研究结果表明,所选益生菌 Lactiplantibacillus 菌株适合在食品和制药行业中用作生物防腐剂起动剂或供人类食用的益生菌。
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来源期刊
Egyptian Pharmaceutical Journal
Egyptian Pharmaceutical Journal PHARMACOLOGY & PHARMACY-
CiteScore
1.10
自引率
0.00%
发文量
37
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