2013 ; 23(7):
|Author||Youngdeuk Lee, Chulhong Oh, Mahanama De Zoysa, Hyowon Kim, Wickramaarachchige Don Niroshana Wickramaarachchi, Ilson Whang, Do-Hyung Kang, Jehee Lee|
|Affiliation||Korea Institute of Ocean Science Technology, Ansan 426-744, Republic of Korea ,Department of Marine Life Sciences, Jeju National University, 66 Jejudaehakno, Ara-Dong, Jeju 690-756, Republic of Korea|
|Title||Molecular Cloning, Overexpression, and Enzymatic Characterization of Glycosyl Hydrolase Family 16 β-Agarase from Marine Bacterium Saccharophagus sp. AG21 in Escherichia coli|
J. Microbiol. Biotechnol.2013 ; 23(7):
|Abstract||An agar-degrading bacterium was isolated from red seaweed (Gelidium amansii) on a natural
seawater agar plate, and identified as Saccharophagus sp. AG21. The β-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) β-agarase module and two carbohydrate binding
modules of family 6 (CBM6). The deduced aa sequence showed 93.7% and 84.9% similarity to
β-agarase of Saccharophagus degradans and Microbulbifer agarilyticus, respectively. The mature
agy1 was cloned and overexpressed as a His-tagged recombinant β-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 55oC and pH 7.6, and had a specific activity of 85 U/mg.
The rAgy1 activity was enhanced by FeSO4 (40%), KCl (34%), and NaCl (34%), compared with
the control. The newly identified rAgy1 is a β-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.|
|Keywords||Saccharophagus sp.AG21, beta-agarase, GH16, neoagaro-oligosaccharide|
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