Molecular Cloning and Enzymatic Characterization of GH16 beta-agarase from Marine Bacterium

Title
Molecular Cloning and Enzymatic Characterization of GH16 beta-agarase from Marine Bacterium
Author(s)
이영득; 권영경; 김주연; 임송이; 박아름이; 오철홍; 강도형
KIOST Author(s)
Lee, Young Deuk(이영득)Park, Areumi(박아름이)Oh, Chul Hong(오철홍)Kang, Do Hyung(강도형)
Alternative Author(s)
이영득; 권영경; 김주연; 임송이; 박아름이; 오철홍; 강도형
Publication Year
2013-05-06
Abstract
A novel β-agarase gene was identified in a Saccharophagus sp. AG21 isolate from the Jeju Island coastal evironment, designated as agy1. Subsequent cloning and expression of the recombinant β-agarase(rAgy1) was analyzed to detemine its biochemical properties. The predicted sequence has 1908 bp ORF encoding 636 amino acids, and includes a glycosyl hydrolase 16 β-agarase module and two carbohydrate binding modules of family 6. The deduced amino acid seqeunce showed 93.7% similarity to beta-agarase of Saccharophagus degradans. The mature agy1 was cloned and overexpressed as a His-tagged recombinant beta-agarase (rAgy1) in Escherichia coli, and had a predicted molecular weight of 69 kDa and an Isoelectric point of 4.5. rAgy1 showed optimum activity at 55℃ and pH 7.5, and had a specific activity of 85U/mg. The rAgy1 activity was enhanced by FeSO4 (40%), KCl (34%) and NaCl (34%). The newly identified rAgy1 is a β-agarase, which acts to degrade agarose to neoagarotetraose and neoagarohexaose and may be useful for applications in the cosmetics, food and reagent industries. Supported by KIOST and MLTM (PE98931, PM57210, PE98932). biochemical properties. The predicted sequence has 1908 bp ORF encoding 636 amino acids, and includes a glycosyl hydrolase 16 β-agarase module and two carbohydrate binding modules of family 6. The deduced amino acid seqeunce showed 93.7% similarity to beta-agarase of Saccharophagus degradans. The mature agy1 was cloned and overexpressed as a His-tagged recombinant beta-agarase (rAgy1) in Escherichia coli, and had a predicted molecular weight of 69 kDa and an Isoelectric point of 4.5. rAgy1 showed optimum activity at 55℃ and pH 7.5, and had a specific activity of 85U/mg. The rAgy1 activity was enhanced by FeSO4 (40%), KCl (34%) and NaCl (34%). The newly identified rAgy1 is a β-agarase, which acts to degrade agarose to neoagarotetraose and neoagarohexaose and may be useful for applications in the cosmetics, food and reagent industries. Supported by KIOST and MLTM (PE98931, PM57210, PE98932).
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/27092
Bibliographic Citation
2013년도 한국미생물학회 국제학술대회, pp.282, 2013
Publisher
한국미생물학회
Type
Conference
Language
English
Publisher
한국미생물학회
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