کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10756967 | 1050390 | 2013 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A novel multifunctional cellulolytic enzyme screened from metagenomic resources representing ruminal bacteria
ترجمه فارسی عنوان
یک آنزیم سلولولویتیک چند منظوره جدید که از منابع متابولیتی نشان داده شده باکتری های شومینه است
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
چکیده انگلیسی
Metagenomic resources representing ruminal bacteria were screened for novel exocellulases using a robotic, high-throughput screening system, the novel CelEx-BR12 gene was identified and the predicted CelEx-BR12 protein was characterized. The CelEx-BR12 gene had an open reading frame (ORF) of 1140 base pairs that encoded a 380-amino-acid-protein with a predicted molecular mass of 41.8 kDa. The amino acid sequence was 83% identical to that of a family 5 glycosyl hydrolase from Prevotella ruminicola 23. Codon-optimized CelEx-BR12 was overexpressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The Michaelis-Menten constant (Km value) and maximal reaction velocity (Vmax values) for exocellulase activity were 12.92 μM and 1.55 Ã 10â4 μmol minâ1, respectively, and the enzyme was optimally active at pH 5.0 and 37 °C. Multifunctional activities were observed against fluorogenic and natural glycosides, such as 4-methylumbelliferyl-β-d-cellobioside (0.3 U mgâ1), CMC (105.9 U mgâ1), birch wood xylan (132.3 U mgâ1), oat spelt xylan (67.9 U mgâ1), and 2-hydroxyethyl-cellulose (26.3 U mgâ1). Based on these findings, we believe that CelEx-BR12 is an efficient multifunctional enzyme as endocellulase/exocellulase/xylanase activities that may prove useful for biotechnological applications.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 441, Issue 3, 22 November 2013, Pages 567-572
Journal: Biochemical and Biophysical Research Communications - Volume 441, Issue 3, 22 November 2013, Pages 567-572
نویسندگان
Kyong-Cheol Ko, Ju Hee Lee, Yunjon Han, Jong Hyun Choi, Jae Jun Song,