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Cloning and Expression Analysis of Xylanase Gene Cluster from Yak Rumen

Author: WangMin
Tutor: TianRuiHua；DaiXin
School: Inner Mongolia Agricultural University
Course: Fermentation Engineering
Keywords: Rumen Metagenome Xylan degrading Gene / gene cluster Synergy
CLC: Q78
Type: Master's thesis
Year: 2011
Downloads: 55
Quote: 0
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Abstract


The rumen excellent biological systems is the nature Celluloytic strongest one. The microbes in this environment provides a great resource for us to develop new genes and enzymes, therefore, the development and utilization of biomass energy research hotspot. The traditional means of research is dependent culture lignocellulose degrading bacteria isolated from the rumen. According to the study of microbial count, 75% of prokaryotes uncultured microorganisms in the rumen. Therefore, plays a very important role in the process of uncultured microorganisms may xylan degradation in the rumen. Genome / per genome technology is one kind of independent in pure culture microbial populations gene group learning the analysis tools, by constructing a yak rumen uncultured bacteria of metagenomic library, directly from the found a large number of wood cellulose degradation enzyme genes / gene clusters, from molecules level to study its mechanism of degradation of lignocellulose to provide strong protection for the development of rumen microbial resources. This study from our laboratory pre build good yak rumen genome BAC library of 9600 clones were screened lignocellulose degrading enzyme gene / gene cluster, 121 (1.26%) xylanase-positive clones, in combination with other enzymes live a total of 293 (3.05%) positive clones. Analysis of these clones using the 454 sequencing, wherein the xylanase and xylosidase enzymes xylan degradation related enzymes, and the GenBank protein database, KEGG database and COG database than the combined Pfam analysis results, and carbon-water compounds the active enzyme analyze site CAZy (http://www.cazy.org) information, screening related gene encoding xylanase enzyme. From screening to uncultured microbial xylanase cloning and expression of a possible xylan degradation gene cluster of related genes. Were analyzed for their enzymatic properties of these enzymes, the optimum reaction temperatures are 40 ° C, the optimum pH is 6, the enzymatic reaction conditions of the rumen environment with which they are consistent. Vitro study of the synergistic effect in the degradation of xylan, the results show the α-glucuronic acid role of the enzyme on the xylanase is very obvious, and also has a role in promoting xylosidase xylanase The lignocellulosic laid the foundation for further study of the the xylan degradation process mechanism and development.

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CLC: > Biological Sciences > Molecular Biology > Genetic engineering (genetic engineering)
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