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Molecular Cloning, Overexpression, and Enzymatic Characterization of Glycosyl Hydrolase Family 16 ${eta}$-Agarase from Marine Bacterium Saccharophagus sp. AG21 in Escherichia coli
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  • Molecular Cloning, Overexpression, and Enzymatic Characterization of Glycosyl Hydrolase Family 16 ${eta}$-Agarase from Marine Bacterium Saccharophagus sp. AG21 in Escherichia coli
  • Molecular Cloning, Overexpression, and Enzymatic Characterization of Glycosyl Hydrolase Family 16 ${eta}$-Agarase from Marine Bacterium Saccharophagus sp. AG21 in Escherichia coli
저자명
Lee. Youngdeuk,Oh. Chulhong,Zoysa. Mahanama De,Kim. Hyowon,Wickramaarachchi. Wickramaarachchige Don Niroshana,Whang. Ilson,Kang.
간행물명
Journal of microbiology and biotechnology
권/호정보
2013년|23권 7호|pp.913-922 (10 pages)
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한국미생물생명공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

An agar-degrading bacterium was isolated from red seaweed (Gelidium amansii) on a natural seawater agar plate, and identified as Saccharophagus sp. AG21. The ${eta}$-agarase gene from Saccharophagus sp. AG21 (agy1) was screened by long and accurate (LA)-PCR. The predicted sequence has a 1,908 bp open reading frame encoding 636 amino acids (aa), and includes a glycosyl hydrolase family 16 (GH16) ${eta}$-agarase module and two carbohydrate binding modules of family 6 (CBM6). The deduced aa sequence showed 93.7% and 84.9% similarity to ${eta}$-agarase of Saccharophagus degradans and Microbulbifer agarilyticus, respectively. The mature agy1 was cloned and overexpressed as a His-tagged recombinant ${eta}$-agarase (rAgy1) in Escherichia coli, and had a predicted molecular mass of 69 kDa and an isoelectric point of 4.5. rAgy1 showed optimum activity at $55^{circ}C$ and pH 7.6, and had a specific activity of 85 U/mg. The rAgy1 activity was enhanced by $FeSO_4$ (40%), KCl (34%), and NaCl (34%), compared with the control. The newly identified rAgy1 is a ${eta}$-agarase, which acts to degrade agarose to neoagarotetraose (NA4) and neoagarohexaose (NA6) and may be useful for applications in the cosmetics, food, bioethanol, and reagent industries.