Use of Metagenomics and Isolation of Actinobacteria in Brazil's Atlantic Rainforest Soil for Antimicrobial Prospecting.

ISRN biotechnology Pub Date : 2014-03-12 eCollection Date: 2014-01-01 DOI:10.1155/2014/909601
Danyelle Alves Martins Assis, Rachel Passos Rezende, João Carlos Teixeira Dias
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引用次数: 6

Abstract

Modern techniques involving molecular biology, such as metagenomics, have the advantage of exploiting a higher number of microorganisms; however, classic isolation and culture methods used to obtain antimicrobials continue to be promising, especially in the isolation of Actinobacteria, which are responsible for the production of many of these compounds. In this work, two methodologies were used to search for antimicrobial substances-isolation of Actinobacteria and metagenomics of the Atlantic Rainforest soil and of the cultivation of cocoa intercropped with acai berry in the Atlantic Rainforest. The metagenomic libraries were constructed with the CopyControl Fosmid Library kit EPICENTRE, resulting in a total of 2688 clones, 1344 of each soil sample. None of the clones presented antimicrobial activity against the microorganisms tested: S. aureus, Bacillus subtilis, and Salmonella choleraesuis. A total of 46 isolates were obtained from the isolation of soil Actinobacteria: 24 isolates from Atlantic Rainforest soil and 22 isolates from the intercrop cultivation soil. Of these, two Atlantic Rainforest soil isolates inhibited the growth of S. aureus including a clinical isolate of S. aureus MRSA-a promising result, since it is an important multidrug-resistant human pathogen.

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利用宏基因组学和分离巴西大西洋雨林土壤放线菌进行抗菌勘探。
涉及分子生物学的现代技术,如宏基因组学,具有利用更多数量微生物的优势;然而,用于获得抗菌剂的经典分离和培养方法仍然很有前途,特别是在分离放线菌方面,放线菌负责产生许多这些化合物。本研究采用两种方法寻找抗微生物物质:大西洋雨林土壤放线菌分离和宏基因组学,以及大西洋雨林可可间作巴西莓种植。使用CopyControl Fosmid Library kit EPICENTRE构建宏基因组文库,共获得2688个克隆,每个土壤样品1344个。这些克隆对金黄色葡萄球菌、枯草芽孢杆菌和霍乱沙门氏菌均无抗菌活性。从土壤中分离得到46株放线菌,其中大西洋雨林土壤分离得到24株放线菌,间作土壤分离得到22株放线菌。其中,两种大西洋雨林土壤分离物抑制了金黄色葡萄球菌的生长,包括金黄色葡萄球菌mrsa的临床分离物——这是一个很有希望的结果,因为它是一种重要的多重耐药人类病原体。
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