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贝加尔湖的附石生物膜:异养细菌基因组中聚酮合酶(PKS)和非核糖体肽合成酶(NRPS)基因的筛选与多样性

Epilithic Biofilms in Lake Baikal: Screening and Diversity of PKS and NRPS Genes in the Genomes of Heterotrophic Bacteria.

作者信息

Sukhanova Elena, Zimens Ekaterina, Kaluzhnaya Oksana, Parfenova Valentina, Belykh Olga

机构信息

Limnological Institute of Siberian Branch of Russian Academy of Sciences , Irkutsk , Russia.

出版信息

Pol J Microbiol. 2018;67(4):501-516. doi: 10.21307/pjm-2018-060.

Abstract

A collection of heterotrophic bacteria consisting of 167 strains was obtained from microbial communities of biofilms formed on solid substrates in the littoral zone of Lake Baikal. Based on the analysis of 16S rRNA gene fragments, the isolates were classified to four phyla: Proteobacteria , Firmicutes , Actinobacteria , and Bacteroidetes . To assess their biotechnological potential, bacteria were screened for the presence of PKS (polyketide synthase) and NRPS (non-ribosomal peptide synthetases) genes. PKS genes were detected in 41 strains (25%) and NRPS genes in 73 (43%) strains by PCR analysis. The occurrence of PKS genes in members of the phylum Firmicutes (the genera Bacillus and Paenibacillus ) was 34% and NRPS genes were found in 78%. In Proteobacteria , PKS and NRPS genes were found in 20% and 32%, and in 22% and 22% of Actinobacteria , respectively. For further analysis of PKS and NRPS genes, six Bacillus and Paenibacillus strains with antagonistic activity were selected and underwent phylogenetic analysis of 16S rRNA genes. The identification of PKS and NRPS genes in the strains investigated was demonstrated among the homologues the genes involved in the biosynthesis of antibiotics (bacillaene, difficidine, erythromycin, bacitracin, tridecaptin, and fusaricidin), biosurfactants (iturin, bacillomycin, plipastatin, fengycin, and surfactin) and antitumor agents (epothilone, calyculin, and briostatin). Bacillus spp. 9A and 2A strains showed the highest diversity of PKS and NRPS genes. Bacillus and Paenibacillus strains isolated from epilithic biofilms in Lake Baikal are potential producers of antimicrobial compounds and may be of practical interest for biotechnological purposes. A collection of heterotrophic bacteria consisting of 167 strains was obtained from microbial communities of biofilms formed on solid substrates in the littoral zone of Lake Baikal. Based on the analysis of 16S rRNA gene fragments, the isolates were classified to four phyla: , , , and . To assess their biotechnological potential, bacteria were screened for the presence of PKS (polyketide synthase) and NRPS (non-ribosomal peptide synthetases) genes. PKS genes were detected in 41 strains (25%) and NRPS genes in 73 (43%) strains by PCR analysis. The occurrence of PKS genes in members of the phylum (the genera and ) was 34% and NRPS genes were found in 78%. In , PKS and NRPS genes were found in 20% and 32%, and in 22% and 22% of , respectively. For further analysis of PKS and NRPS genes, six and strains with antagonistic activity were selected and underwent phylogenetic analysis of 16S rRNA genes. The identification of PKS and NRPS genes in the strains investigated was demonstrated among the homologues the genes involved in the biosynthesis of antibiotics (bacillaene, difficidine, erythromycin, bacitracin, tridecaptin, and fusaricidin), biosurfactants (iturin, bacillomycin, plipastatin, fengycin, and surfactin) and antitumor agents (epothilone, calyculin, and briostatin). spp. 9A and 2A strains showed the highest diversity of PKS and NRPS genes. and strains isolated from epilithic biofilms in Lake Baikal are potential producers of antimicrobial compounds and may be of practical interest for biotechnological purposes.

摘要

从贝加尔湖沿岸带固体基质上形成的生物膜微生物群落中获得了一组由167个菌株组成的异养细菌。基于对16S rRNA基因片段的分析,这些分离株被归类为四个门:变形菌门、厚壁菌门、放线菌门和拟杆菌门。为了评估它们的生物技术潜力,对细菌进行了聚酮合酶(PKS)和非核糖体肽合成酶(NRPS)基因存在情况的筛选。通过PCR分析,在41个菌株(25%)中检测到PKS基因,在73个菌株(43%)中检测到NRPS基因。厚壁菌门成员(芽孢杆菌属和类芽孢杆菌属)中PKS基因的出现率为34%,NRPS基因的出现率为78%。在变形菌门中,PKS和NRPS基因的出现率分别为20%和32%,在放线菌门中分别为22%和22%。为了进一步分析PKS和NRPS基因,选择了6个具有拮抗活性的芽孢杆菌和类芽孢杆菌菌株,并对其16S rRNA基因进行了系统发育分析。在所研究菌株中鉴定出的PKS和NRPS基因与参与抗生素(杆菌烯、艰难梭菌素、红霉素、杆菌肽、十三肽菌素和镰刀菌素)、生物表面活性剂(伊枯草菌素、芽孢杆菌霉素、脂肽菌素、丰原素和表面活性素)和抗肿瘤剂(埃坡霉素、花萼海绵诱癌素和苔藓抑素)生物合成的基因同源。芽孢杆菌属的9A和2A菌株显示出PKS和NRPS基因的最高多样性。从贝加尔湖石上生物膜分离出的芽孢杆菌和类芽孢杆菌菌株是抗菌化合物的潜在生产者,可能在生物技术方面具有实际应用价值。从贝加尔湖沿岸带固体基质上形成的生物膜微生物群落中获得了一组由167个菌株组成的异养细菌。基于对16S rRNA基因片段的分析,这些分离株被归类为四个门: 、 、 和 。为了评估它们的生物技术潜力,对细菌进行了聚酮合酶(PKS)和非核糖体肽合成酶(NRPS)基因存在情况的筛选。通过PCR分析,在41个菌株(25%)中检测到PKS基因,在73个菌株(43%)中检测到NRPS基因。 门成员( 属和 属)中PKS基因的出现率为34%,NRPS基因的出现率为78%。在 中,PKS和NRPS基因的出现率分别为20%和32%,在 的22%和22%中分别发现。为了进一步分析PKS和NRPS基因,选择了6个具有拮抗活性的 和 菌株,并对其16S rRNA基因进行了系统发育分析。在所研究菌株中鉴定出的PKS和NRPS基因与参与抗生素(杆菌烯、艰难梭菌素、红霉素、杆菌肽、十三肽菌素和镰刀菌素)、生物表面活性剂(伊枯草菌素、芽孢杆菌霉素、脂肽菌素、丰原素和表面活性素)和抗肿瘤剂(埃坡霉素、花萼海绵诱癌素和苔藓抑素)生物合成的基因同源。 属的9A和2A菌株显示出PKS和NRPS基因的最高多样性。从贝加尔湖石上生物膜分离出的 和 菌株是抗菌化合物的潜在生产者,可能在生物技术方面具有实际应用价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b542/7256756/aaf6ff13b0a0/pjm-67-4-501-g001.jpg

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