β-agarase from Pseudomonas sp. W7 : purification of the recombinant enzyme from Escherichia coli and the effects of salt on its activity

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Title
β-agarase from Pseudomonas sp. W7 : purification of the recombinant enzyme from Escherichia coli and the effects of salt on its activity
Author(s)
Jeong-Chul Ha; Gu-Taek Kim; Sung-Koo Kim; Tae Kwang Oh; Ju-Hyun Yu; In-Soo Kong
Bibliographic Citation
Biotechnology and Applied Biochemistry, vol. 26, no. 1, pp. 1-6
Publication Year
1997
Abstract
The recombinant plasmid (pJAI), harbouring the agarase gene (pjaA) of Pseudomonas sp. W7, was introduced and expressed in Escherichia coli JM83. The agarase was purified using a combination of acetone precipitation and anion-exchange, gel-filtration and affinity chromatographies, with overall yield of 10% from the culture supernatant of E. coli JM83 (pJAI). The purified agarase migrated as a single band (molecular mass 59 kDa) on SDS/PAGE and was found to be β-agarase, which could hydrolyse the β-1,4 linkage of agarose to yield neoagarotetraose as the main product. Optimal enzyme activity was at pH 7.8 and the temperature optimum spanned the broad range 20-40 °C. The recombinant agarase was halophilic, maximum activity being exhibited at 0.9 M NaCl. This halophilic property could improve the production of neoagaro-oligosaccharides available in a marine environment.
Keyword
beta agaraserecombinant enzymeenzyme activityenzyme purificationEscherichia coliPseudomonasrecombinant proteins
ISSN
0885-4513
Publisher
Wiley
Type
Article
Appears in Collections:
Division of Biomedical Research > Metabolic Regulation Research Center > 1. Journal Articles
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