Extracting method of cellulose degradation flora metagenome

Abstract

The invention discloses an extracting method of cellulose degradation flora metagenome and provides an extracting method of cellulose degradation flora metagenome DNA. The extracting method comprises the following steps that cellulose degradation florae are sequentially subjected to thallus wall breaking, DNA organic solution extraction, DNA isoamylol sediment and DNA solution so that the metagenome DNA can be obtained. Experiments prove that a high-salt solution is adopted for carrying out eluting before extraction, lysozyme and mechanical homogenate are added to reinforce cell wall breaking, proteinase K is adopted for wall breaking, cell walls of microorganisms can be broken more easily, and DNA molecules can be released more easily. Meanwhile, the process repeatability is effectively ensured, and then integrity and purity of extracted metagenome DNA fragments and flora structure integrity are ensured. The extracting method provides an effective technology support for study and application of the natural cellulose utilizing florae and the microorganism molecular ecology study.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting the metagenomic group of cellulose-degrading bacteria. Background technique [0002] Lignocellulose is the most productive polysaccharide in nature, and it widely exists in forests, grasslands and farmlands. Its output is abundant, its price is low, and it mostly exists in the form of agricultural and industrial waste, so it is an ideal raw material for the production of bioenergy substances. Studies have shown that the bioenergy production process using lignocellulose as raw material can not only realize the reuse of agricultural waste, but also effectively avoid the contradiction between energy production and food resources, and has become the most potential way of industrialized bioenergy production. [0003] However, due to the complex composition and compact structure of lignocellulose, the process of hydrolyzing it into usable sugars requires the synerg...

AI Technical Summary

Problems solved by technology

However, due to the characteristics of the cellulosic flora, it is difficult to extract its genomic DNA. Therefore, a complete macrogene analysis technology for the cellulosic flora has not yet been established.

[0005] The factors affecting the extraction effect of the metagenomics of the natural cellulose flora mainly include the following three points: First, the strains in the cellulose flora are mostly tightly combined with the cellulose microfilaments through the structure of the cellulosus. The cross-linking form of "body" exists in the fermentation broth, which will significantly affect the breaking of the cell wall and the extraction of the genome, resulting in the fact that the obtained DNA cannot truly reflect the composition of the flora and affect the accuracy of the analysis of the flora structure; the second During the process of bacterial fermentation of cellulose, a large number of metabolites will accumulate outside the cells, including organic acids, organic alcohols and surfactants. These substances will interfere with the genome extraction reagents and affect DNA extraction. Effect; third, the flora contains both Gram-negative bacteria and Gram-positive bacteria, and there are a large number of spores, which increases the difficulty of DNA extraction

Images