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巴氏杆菌属(LMA4)的体外生物活性和候选基因的计算机注释。

Bioactivity of Microbacterium barkeri (LMA4) In Vitro and Candidate Gene Annotation In Silico.

机构信息

Laboratory of Medical Microbiology, Department of Biotechnology and Bioinformatics, School of Life Sciences, Sambalpur University, Jyoti Vihar, Burla, 768019, Odisha, India.

出版信息

Appl Biochem Biotechnol. 2024 May;196(5):2839-2850. doi: 10.1007/s12010-023-04519-5. Epub 2023 May 11.

Abstract

Actinomycetes are considered a never-ending treasure trove of biometabolites, which always fascinated researchers. However, to combat with newly emerging bacterial strains, the search for novel or analogs of existing therapeutic agents is recommended. In this context, this research work was carried out to search for a biopotent Actinomycetal strain grown in untapped soil, near the Hirakud dam. This Gram-positive bacteria was subjected to screening for its bioactivity against the medically important bacteria, isolated from local hospital sample using "co-culture" method, following both qualitative and quantitative assays. Further, the 16 s rRNA sequencing, BLASTn analysis, and GC% calculation were carried out. Based upon its bioactivity, a prediction-based genomics work was pursued, considering the gene sequence deposited in public domain. The reverse translation, elution of protein structural file, and the putative protein were predicted. The strain was identified as Microbacterium barkeri, with 54.1% GC content. From Gene Ontology term annotation, it was predicted that the α/β hydrolase fold of hydrolase protein could have been responsible for antibiotic/biometabolite synthesis, in silico. The in vitro-based sequence (from Whole Genome Sequence data) had inferred that there was elution of alpha/beta hydrolase fold, substantiated with conserved domain analysis, ORF finding more over Gene Ontology (GO) terminology annotations. The GO annotations had suggested that the protein had been produced in response to a bacteria, under the influence of external stimuli more so in stress.

摘要

放线菌被认为是生物代谢物的不竭宝库,一直吸引着研究人员。然而,为了应对新出现的细菌菌株,建议寻找新的或现有治疗剂的类似物。在这种情况下,进行了这项研究工作,以寻找在未开发土壤中生长的具有生物潜力的放线菌菌株,该土壤位于 Hirakud 大坝附近。这种革兰氏阳性细菌通过“共培养”方法,针对从当地医院样本中分离出的具有医学重要性的细菌进行了生物活性筛选,采用定性和定量测定。此外,还进行了 16s rRNA 测序、BLASTn 分析和 GC%计算。根据其生物活性,考虑到公共领域中已存入的基因序列,进行了基于预测的基因组学工作。进行了反向翻译、蛋白结构文件的洗脱和推定蛋白的预测。该菌株被鉴定为 Microbacterium barkeri,GC 含量为 54.1%。从基因本体论术语注释来看,预测水解酶蛋白的α/β水解酶折叠可能负责抗生素/生物代谢物的合成,这是在计算机上完成的。基于体外的序列(来自全基因组序列数据)推断出洗脱了 α/β 水解酶折叠,保守结构域分析、ORF 发现以及基因本体论 (GO) 术语注释进一步证实了这一点。GO 注释表明,该蛋白是在外部刺激(尤其是在压力下)的影响下,针对细菌产生的。

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