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Purification and biochemical characterization of extracellular glucoamylase from Paenibacillus amylolyticus strain
Author(s) -
Lincoln Lynette,
More Veena S.,
More Sunil S.
Publication year - 2019
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201800540
Subject(s) - hydrolysis , size exclusion chromatography , chemistry , chromatography , starch , paenibacillus , extracellular , fermentation , enzyme , biochemistry , substrate (aquarium) , biology , ecology , 16s ribosomal rna , gene
In the present study, glucoamylase produced from a soil bacterium Paenibacillus amylolyticus NEO03 was cultured under submerged fermentation conditions. The extracellular enzyme was purified by starch adsorption chromatography and further by gel filtration, with 2.73‐fold and recovery of 40.02%. The protein exhibited molecular mass of ∼66,000 Da as estimated by SDS–PAGE and depicted to be a monomer. The enzyme demonstrated optimum activity at pH range 6.0–7.0 and temperature range 30–40 °C. Glucoamylase was mostly activated by Mn 2+ metal ions and depicted no dependency on Ca 2+ ions. The enzyme preferentially hydrolyzed all the starch substrates. High substrate specificity was demonstrated towards soluble starch and kinetic values K m and V max were 2.84 mg/ml and 239.2 U/ml, respectively. The products of hydrolysis of soluble starch were detected by thin layer chromatography which showed only D ‐glucose, indicating a true glucoamylase. The secreted glucoamylase from P. amylolyticus strain possesses properties suitable for saccharification processes such as biofuel production.