کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5906720 1159983 2013 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A metagenomic approach for the identification and cloning of an endoglucanase from rice straw compost
ترجمه فارسی عنوان
یک رویکرد مکتب زیستی برای شناسایی و شبیه سازی اندگلوکاناز از کمپوست نیترات برنج
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی ژنتیک
چکیده انگلیسی

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.

► A novel endoglucanase gene was cloned from DNA isolated from rice straw compost. ► Less than 3000 clones were screened to isolate the RS-EG1 endoglucanase gene. ► RS-EG1 shares about 70% similarity with the closest known bacterial cellulases. ► RS-EG1 is stable over broad temperature and pH ranges. ► RS-EG1 is potentially useful in cellulosic biofuel production.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 519, Issue 2, 1 May 2013, Pages 360-366
نویسندگان
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