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An agar-degrading bacterium, strain BN3, was isolated from a coastal soil sample collected in Taiwan Strait, China and identified to be a novel species of the genus Microbulbifer. The gene (N3-1) encoding for a novel beta-agarase from the isolate was cloned and sequenced. It encoded a mature protein with 274 amino acids and a calculated molecular mass of 34.3 kDa. The deduced amino acid sequence manifested sequence similarity (61-84% identity) to characterized beta-agarases in the glycoside hydrolase family 16. The recombinant agarase was hyper-produced extracellularly using Pichia pastoris as the host. After induction in a shake flask for 96 h, the yield of recombinant N3-1 protein reached 0.406 mg/mL, and the enzyme activity attained 502.1 U/mL. The enzyme purified by ion exchange chromatography displayed a specific activity of 6447 U/mg at pH 6.0 and 50 degrees C. The optimal pH and temperature for agarase activity were approximately 6 and 50 degrees C, respectively. The pattern of agarose hydrolysis showed that the enzyme was an endo-type beta-agarase, capable of hydrolyzing agarose and Gracilaria lemaneiformis, with neoagarobiose and neoagarotetraose as the final main products. (C) 2018 Elsevier B.V. All rights reserved.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
ISSN: 0141-8130
Year: 2018
Volume: 119
Page: 1164-1170
4 . 7 8 4
JCR@2018
7 . 7 0 0
JCR@2023
ESI Discipline: BIOLOGY & BIOCHEMISTRY;
ESI HC Threshold:212
JCR Journal Grade:1
CAS Journal Grade:2
Cited Count:
SCOPUS Cited Count:
ESI Highly Cited Papers on the List: 0 Unfold All
WanFang Cited Count:
Chinese Cited Count:
30 Days PV: 1
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