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Ramos, Kristine Rose M.,Valdehuesa, Kris Niñ,o G.,Maza, Perry Ayn Mayson M.,Nisola, Grace M.,Lee, Won-Keun,Chung, Wook-Jin Elsevier Science B.V., Amsterdam. 2017 PROCESS BIOCHEMISTRY Vol.63 No.-
<P><B>Abstract</B></P> <P>An α-neoagarobiose hydrolase (α-NABH) from <I>Cellulophaga</I> sp. W5C, designated as AhgI, was identified, purified, and characterized. Its 1227 base pairs of coded sequence translate into a 408-amino acid protein that belongs to the GH117 family. Multiple sequence alignment of AhgI with other known α-NABHs showed 83% homology with AhgA from <I>Zobellia galactanivorans</I>. AhgI had an apparent molecular weight of 45kDa and was highly active at pH 7.0 and 20°C. The <I>K</I> <SUB>m</SUB> and <I>V</I> <SUB>max</SUB> values for neoagarobiose (NA2) were 1.03mM and 10.22U/mg, respectively. Apart from NA2, the enzyme showed activity against other neoagaro-oligosaccharides such as neoagarotetraose (NA4) and neoagarohexaose (NA6). AhgI was then employed in a prototype process to produce D-galactonate from <I>Gelidium amansii</I>. Agar from <I>G. amansii</I> was hydrothermally extracted and then enzymatically hydrolyzed by sequential addition of β-agarases and AhgI. The final hydrolysate containing D-galactose was then utilized for the microbial production of D-galactonate. This is believed to be the first report on the identification and characterization of an α-NABH derived from <I>Cellulophaga</I> species and its subsequent application in the synthesis of a value-added chemical directly from marine macroalgae.</P> <P><B>Highlights</B></P> <P> <UL> <LI> α-Neoagarobiose hydrolase was identified from <I>Cellulophaga</I> sp. W5C. </LI> <LI> Optimum conditions for enzyme activity are pH 7.0 and 20°C. </LI> <LI> AhgI was active towards neoagarobiose, neoagarotetraose, and neoagarohexaose. </LI> <LI> AhgI was applied for the hydrolysis of agar and subsequent D-galactonate production. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
Isolation and Characterization of a Novel Marine Bacterium Belonging to Cellulophaga Genus
Kristine Rose M. RAMOS,Kris Nino G. VALDEHUESA,Perry Ayn Mayson M. MAZA,Teklebrahan Gebrekrstos WELDEM,Angelo B. BAnARES,Rhudith B. CABULONG,Grace M. NISOLA,Won Keun LEE,Wook Jin CHUNG 한국생물공학회 2016 한국생물공학회 학술대회 Vol.2016 No.10