NDLI: A novel bifunctional endo-/exo-type cellulase from an anaerobic ruminal bacterium

Content Provider	Springer Nature Link
Author	Ko, Kyong Cheol Han, Yunjon Choi, Jong Hyun Kim, Geun Joong Lee, Seung Goo Song, Jae Jun
Copyright Year	2010
Abstract	An anaerobic microorganism termed AN-C16-KBRB was isolated from the bovine rumen and demonstrated cellulolytic activity on a NB agar plate containing azo-carboxymethyl cellulose. The 16S rRNA gene of the strain was 98% similar to that of Clostridiaceae bacterium SK082 (AB298754) as the highest homology. A novel celEdx16 gene encoding a bifunctional endo-/exocellulase (CelEdx16) was cloned by the shotgun method from AN-C16-KBRB, and the enzyme was characterized. The celEdx16 gene had an open reading frame of 1,104-base pairs, which encoded 367 amino acids to yield a protein of molecular mass 40.4 kDa. The amino acid sequence was 53% identical to that of an endoglucanase from Clostridium thermocellum. CelEdx16 was overexpressed in Escherichia coli and purified using Ni-NTA affinity chromatography. The specific endocellulase and exocellulase activities of CelEdx16 were 15.9 and 3.6 × 10−2 U mg−1, respectively. The Michaelis–Menten constant (K m values) and the maximal reaction velocities (V max values) of CelEdx16 were 47.1 μM and 9.6 × 10−3 μmole min−1 when endocellulase activity was measured and 106.3 μM and 2.1 × 10−5 μmole min−1 when exocellulase activity was assessed. CelEdx16 was optimally active at pH 5.0 and 40°C.
Starting Page	1453
Ending Page	1462
Page Count	10
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	89
Issue Number	5
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2010-11-03
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Screening Korean bovine ruminal bacterium Bifunctional endo-/exo-type cellulase Cloning Expression Microbial Genetics and Genomics Microbiology Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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