Title: | Direct production of ethanol from neoagarobiose using recombinant yeast that secretes alpha-neoagarooligosaccharide hydrolase |
Author(s): | Syazni; Yanagisawa M; Kasuu M; Ariga O; Nakasaki K; |
Address: | "Department of International Development Engineering, Graduate School of Engineering, Tokyo Institute of Technology, 2-12-1 I4-2 Ookayama, Meguro-ku, Tokyo 152-8550, Japan. Department of Environmental Sciences, School of Food, Agricultural and Environmental Sciences, Miyagi University, 2-2-1 Hatadate, Taihaku-ku, Sendai City, Miyagi 982-0215, Japan. School of Environmental Science and Engineering, Kochi University of Technology, 185 Miyanokuchi, Kami City, Kochi 782-8502, Japan. Department of International Development Engineering, Graduate School of Engineering, Tokyo Institute of Technology, 2-12-1 I4-2 Ookayama, Meguro-ku, Tokyo 152-8550, Japan. Electronic address: nakasaki@ide.titech.ac.jp" |
DOI: | 10.1016/j.enzmictec.2015.08.013 |
ISSN/ISBN: | 1879-0909 (Electronic) 0141-0229 (Linking) |
Abstract: | "An alpha-neoagarooligosaccharide hydrolase, AgaNash, was purified from Cellvibrio sp. OA-2007, which utilizes agarose as a substrate. The agaNash gene, which encodes AgaNash, was obtained by comparing the N-terminal amino acid sequence of AgaNash with that deduced from the nucleotide sequence of the full-length OA-2007 genome. The agaNash gene combined with the Saccharomyces cerevisiae signal peptide alpha-mating factor was transformed into the YPH499 strain of S. cerevisiae to generate YPH499/pTEF-MF-agaNash, and the recombinant yeast was confirmed to produce AgaNash, though it was mainly retained within the recombinant cell. To enhance AgaNash secretion from the cell, the signal peptide was replaced with a combination of the signal peptide and a threonine- and serine-rich tract (T-S region) of the S. diastaticus STA1 gene. The new recombinant yeast, YPH499/pTEF-STA1SP-agaNash, was demonstrated to secrete AgaNash and hydrolyze neoagarobiose with an efficiency of as high as 84%, thereby producing galactose, which is a fermentable sugar for the yeast, and ethanol, at concentrations of up to 1.8 g/L, directly from neoagarobiose" |
Keywords: | "Biofuels Cellvibrio/enzymology/genetics Cloning, Molecular Disaccharides/*metabolism Ethanol/*metabolism Fermentation Galactose/metabolism Genes, Bacterial Glycoside Hydrolases/genetics/*metabolism Mating Factor/genetics Protein Sorting Signals/genetics R;" |
Notes: | "MedlineSyazni Yanagisawa, Mitsunori Kasuu, Masahiro Ariga, Osamu Nakasaki, Kiyohiko eng 2016/02/28 Enzyme Microb Technol. 2016 Apr; 85:82-9. doi: 10.1016/j.enzmictec.2015.08.013. Epub 2015 Aug 28" |