Overexpression and characterization of a novel α-neoagarobiose hydrolase and its application in the production of D-galactonate from Gelidium amansii
| Autor: | Grace M. Nisola, Won-Keun Lee, Perry Ayn Mayson M. Maza, Wook-Jin Chung, Kris Niño G. Valdehuesa, Kristine Rose M. Ramos |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine chemistry.chemical_classification Multiple sequence alignment food.ingredient Gelidium amansii Cellulophaga Bioengineering Biology biology.organism_classification 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Hydrolysate 03 medical and health sciences Hydrolysis 030104 developmental biology Enzyme food chemistry 010608 biotechnology Hydrolase Agar |
| Zdroj: | Process Biochemistry. 63:105-112 |
| ISSN: | 1359-5113 |
| DOI: | 10.1016/j.procbio.2017.08.014 |
| Popis: | An α-neoagarobiose hydrolase (α-NABH) from Cellulophaga 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 Zobellia galactanivorans. AhgI had an apparent molecular weight of 45 kDa and was highly active at pH 7.0 and 20 °C. The Km and Vmax values for neoagarobiose (NA2) were 1.03 mM and 10.22 U/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 Gelidium amansii. Agar from G. amansii 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 Cellulophaga species and its subsequent application in the synthesis of a value-added chemical directly from marine macroalgae. |
| Databáze: | OpenAIRE |
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