期刊
GENE
卷 519, 期 2, 页码 360-366出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.gene.2012.07.076
关键词
Metagenomic library; Rice straw composts; Functional screening; Endoglucanase; Industrial application
资金
- National Science Council of the Republic of China [NSC96-3114-P-001-004, NSC97-3114-P-001-001]
Over the years, culturable cellulase-producing microorganisms have been isolated from a variety of sources and genes of cellulolytic enzymes have been cloned. Then again, the great plate count anomaly phenomenon necessitates a culture-independent metagenomic approach for the isolation of cellulolytic genes from microorganisms in their natural environment. We have constructed a metagenomic library derived from rice straw composts. Of 2739 clones screened, a lambda clone carrying a 12 kb genomic fragment was able to yield a clear zone on an agar plate containing carboxymethyl cellulose (CMC). A 4.7 kb subclone, generated by restriction enzyme digestion, was found to harbor a GH12 cellulase gene, RSC-EG1, encoding 464 amino acids along with two other ORFs. The identified cellulolytic gene showed more than 70% similarity on the amino acid level with cellulase from Micromonospora aurantiaca and Thermobispora sp. Interestingly, RSC-EG1 contains a stretch of approximately 86 amino acids not present in either of these close relatives. Our results demonstrated that RSC-EG1, stable over a wide temperature and pH range, is a novel endoglucanase, and provided the first example of metagenomics approach to isolate cellulolytic gene from rice straw composts. (C) 2012 Elsevier B.V. All rights reserved.
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