Genetic and functional characterization of an extracellular modular GH6 endo-β-1,4-glucanase from an earthworm symbiont, Cellulosimicrobium funkei HY-13

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Title
Genetic and functional characterization of an extracellular modular GH6 endo-β-1,4-glucanase from an earthworm symbiont, Cellulosimicrobium funkei HY-13
Author(s)
Do Young Kim; Min Ji Lee; Han Young Cho; J S Lee; M H Lee; C W Chung; D H Shin; Y H Rhee; Kwang- Hee SonHo Yong Park
Bibliographic Citation
Antonie Van Leeuwenhoek International Journal of General and Molecular Microbiology, vol. 109, pp. 1-12
Publication Year
2016
Abstract
The gene (1608-bp) encoding a GH6 endo-β-1,4-glucanase (CelL) from the earthworm-symbiotic bacterium Cellulosimicrobium funkei HY-13 was cloned from its whole genome sequence, expressed recombinantly, and biochemically characterized. CelL (56.0 kDa) is a modular enzyme consisting of an N-terminal catalytic GH6 domain (from Val57 to Pro396), which is 71 % identical to a GH6 protein (accession no.: WP_034662937) from Cellulomonas sp. KRMCY2, together with a C-terminal CBM 2 domain (from Cys429 to Cys532). The highest catalytic activity of CelL toward carboxymethylcellulose (CMC) was observed at 50 °C and pH 5.0, and was relatively stable at a broad pH range of 4.0?10.0. The enzyme was capable of efficiently hydrolyzing the cellulosic polymers in the order of barley β-1,3-1,4-d-glucan > CMC > lichenan > Avicel > konjac glucomannan. However, cellobiose, cellotriose, p-nitrophenyl derivatives of mono- and disaccharides, or structurally unrelated carbohydrate polymers including β-1,3-d-glucan, β-1,4-d-galactomannan, and β-1,4-d-xylan were not susceptible to CelL. The enzymatic hydrolysis of cellopentaose resulted in the production of a mixture of 68.6 % cellobiose and 31.4 % cellotriose but barley β-1,3-1,4-d-glucan was 100 % degraded to cellotriose by CelL. The enzyme strongly bound to Avicel, ivory nut mannan, and chitin but showed relatively weak binding affinity to lichenan, lignin, or poly(3-hydroxybutyrate) granules.
Keyword
Cellulosimicrobium funkei HY-13Earthworm-symbiotic bacteriumEndo-β-1,4-glucanaseGH family 6
ISSN
0003-6072
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s10482-015-0604-2
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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