Xavier A R E O, Lima E R, Oliveira A M E, Cardoso L, Santos J, Cangussu C H C, Leite L N, Quirino M C L, Júnior I G C, Oliveira D A, Xavier M A S
Laboratório de Bioprospecção e Recursos Genéticos, Centro de Ciências Biológicas e da Saúde, Universidade Estadual de Montes Claros, Centro de Ciências Biológicas e da Saúde, Laboratório de Bioprospecção e Recursos Genéticos, , Brasil
Laboratório de Microbiologia, Centro de Ciências Biológicas e da Saúde, , , Brasil
Genet Mol Res. 2017 Sep 27;16(3):gmr-16-03-gmr.16039771. doi: 10.4238/gmr16039771.
The microorganisms are the best source of extracellular enzymes since they allow an economical technology with low-resource consumption compared to animals and plants. The amylases are among the most important enzymes being the genus Bacillus one of the most investigated due to its ability to produce this enzyme. The objective of this study was to isolate and analyze the genetic diversity among bacteria of the genus Bacillus sp producer of amylase originated from the soil. To this end, soil samples were collected and submitted to the condition of extreme temperature. The serial dilution procedure followed by seeding on solid medium containing starch was used for isolation of strains that produce amylase. The microorganisms isolated were subjected to standard morphological methods for presumptive identification of the genus Bacillus. The PCR assay with the universal genetic marker 16S rDNA was used for confirmation of bacterial strain. All the 10 isolates presumptively identified as bacteria amplified a fragment of 370 bp corresponding to the 16S rDNA gene. The enzymatic activity was expressed as an enzymatic index (EI), after 24 h of incubation. All isolate producers of amylase exhibited EI ≥ 2.0. The determination of the genetic profile and the clonal relationship among the isolates were performed by the method of ERIC-PCR polymorphism. The isolates of Bacillus spp were divided into 2 groups (I and II). Through this method, the discriminatory capacity of this analysis of polymorphisms was verified in differing producer strains from those not producing amylase.
微生物是胞外酶的最佳来源,因为与动植物相比,它们能实现资源消耗低的经济技术。淀粉酶是最重要的酶之一,芽孢杆菌属因其产生这种酶的能力而成为研究最多的属之一。本研究的目的是分离和分析源自土壤的淀粉酶产生菌芽孢杆菌属细菌之间的遗传多样性。为此,采集土壤样本并使其处于极端温度条件下。采用连续稀释法,随后在含淀粉的固体培养基上接种,用于分离产生淀粉酶的菌株。对分离出的微生物采用标准形态学方法进行芽孢杆菌属的初步鉴定。使用通用遗传标记16S rDNA的PCR检测来确认细菌菌株。所有初步鉴定为细菌的10个分离株均扩增出一段对应于16S rDNA基因的370 bp片段。孵育24小时后,酶活性以酶指数(EI)表示。所有淀粉酶产生菌分离株的EI均≥2.0。通过ERIC-PCR多态性方法对分离株之间的遗传图谱和克隆关系进行了测定。芽孢杆菌属分离株分为2组(I和II)。通过这种方法,验证了这种多态性分析在区分淀粉酶产生菌株和非产生菌株方面的鉴别能力。