title: Screening of a glucoside 3-dehydrogenase-producing strain, Sphingobacterium faecium, based on a high-throughput screening method and optimization of the culture conditions for enzyme production.
abstract: The objective of this study was to screen glucoside 3-dehydrogenase (G3DH)-producing strain based on a high-throughput G3DH screening method. Optimization of culture conditions of the isolated strain was also applied in this study. This screening method employed electron transfer reaction in 96-well microtiter plates, α-methyl-D-glucoside, galactose, 2-deoxy-D-glucose, and 3-O-methyl-D-glucose were used as substrates. Using this screening method, one out of 78 strains isolated from different soil samples was obtained with high G3DH activity. The accuracy of the screening method was proved by alkaline treatment analysis of 3-keto sugars. The isolated strain was identified as Sphingobacterium faecium ZJF-D6 by phenotypic characterization and 16S rDNA sequence analysis. The culture conditions of S. faecium for G3DH production were optimized. Sucrose was found as the most suitable carbon source for the G3DH production. The highest G3DH production and cell growth were achieved using the medium at the initial pH of 7.0 at 25 °C for 36 h with activity of 8.03 × 10(-2) U/mL culture. This strain appears promising for potential application in the industry to produce 3-keto sugars. To our knowledge, this is the first report on S. faecium for G3DH production. The method described herein represents a useful tool for the high-throughput isolation of G3DH.
authors.affiliation: College of Biology and Environmental Engineering, Zhejiang Shuren University, Hangzhou, 310015, China, grace_zjf@126.com.
authors.forename: Jianfen
authors.initials: J
authors.lastname: Zhang
authors.forename: Weiqing
authors.initials: W
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Ke
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D001431:Bacteriological Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D005948:Glucose Dehydrogenases / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D045249:Sphingobacterium / Q000201:enzymology* / Q000254:growth & development
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D005948:Glucose Dehydrogenases; C051543:glucoside 3-dehydrogenase