RRC ID 34859
Author Mizuno T, Shiono Y, Koseki T.
Title Biochemical characterization of Aspergillus oryzae native tannase and the recombinant enzyme expressed in Pichia pastoris.
Journal J. Biosci. Bioeng.
Abstract In this study, the biochemical properties of the recombinant tannase from Aspegillus oryzae were compared with those of the native enzyme. Extracellular native tannase was purified from a commercial enzyme source. Recombinant tannase highly expressed in Pichia pastoris was prepared as an active extracellular protein. Purified native and recombinant tannases produced smeared bands with apparent molecular masses of 45-80 kDa and 45-75 kDa, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After N-deglycosylation, the native enzyme yielded molecular masses of 33 kDa and 30 kDa, whereas the recombinant enzyme yielded molecular masses of 34 kDa and 30 kDa. Purified native and recombinant tannases had an optimum pH of 4.0-5.0 and 5.0, respectively, and were stable up to 40°C. After N-deglycosylation, both enzymes exhibited reduced thermostability. Catalytic efficiencies of both purified enzymes were greater with natural substrates, such as (-)-catechin, (-)-epicatechin, and (-)-epigallocatechin gallates, than those with synthetic substrates, such as methyl, ethyl, and propyl gallates. However, there were no activities against the methyl esters of ferulic, p-coumaric, caffeic, and sinapic acids, which indicate feruloyl esterase activity, or the ethyl, propyl, and butyl esters of 4-hydroxybenzoic acid, which indicate paraben hydrolase activity.
Volume 118(4)
Pages 392-5
Published 2014-10
DOI 10.1016/j.jbiosc.2014.04.003
PII S1389-1723(14)00134-0
PMID 24856589
MeSH Aspergillus oryzae / chemistry* Aspergillus oryzae / enzymology Carboxylic Ester Hydrolases / chemistry* Carboxylic Ester Hydrolases / genetics Carboxylic Ester Hydrolases / metabolism Catechin / analogs & derivatives Catechin / chemistry Catechin / metabolism Electrophoresis, Polyacrylamide Gel Fungal Proteins / chemistry* Fungal Proteins / genetics Fungal Proteins / metabolism Gene Expression Hydrogen-Ion Concentration Kinetics Molecular Weight Pichia / enzymology* Pichia / genetics Protein Subunits / chemistry* Protein Subunits / genetics Protein Subunits / metabolism Recombinant Proteins / chemistry Recombinant Proteins / genetics Recombinant Proteins / metabolism Substrate Specificity
IF 2.015
Times Cited 5
WOS Category FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Resource
DNA material pPICZalphaC_AotanA (RDB13297)