Abstract:
:The gene encoding a novel glucoamylase (GlucaM) from the Corallococcus sp. strain EGB was cloned and heterologous expressed in Escherichia coli BL21(DE3), and the enzymatic characterization of recombinant GlucaM (rGlucaM) was determined in the study. The glucaM had an open reading frame of 1938 bp encoding GlucaM of 645 amino acids with no signal peptide. GlucaM belongs to glycosyl hydrolase family 15 and shares the highest identity 96% with the GH15 glucoamylase of Corallococcus coralloides DSM 2259. The rGlucaM with His-tag was purified by the Ni2+-NTA resin, with a specific activity from 3.4 U/mg up to 180 U/mg, and the molecular weight of rGlucaM was approximately 73 kDa on SDS-PAGE. The Km and Vmax of rGlucaM for soluble starch were 1.2 mg/mL and 46 U/mg, respectively. rGlucaM was optimally active at pH 7.0 and 50 °C and had highly tolerance to high concentrations of salts, detergents, and various organic solvents. rGlucaM hydrolyzed soluble starch to glucose, and hydrolytic activities were also detected with amylopectin, amylase, glycogen, starch (potato), α-cyclodextrin, starch (corn and potato). The analysis of hydrolysis products shown that rGlucaM with α-(1-4),(1-6)-D-glucan glucohydrolase toward substrates. These characteristics indicated that the GlucaM was a new member of glucoamylase family and a potential candidate for industrial application.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Li Z,Ji K,Dong W,Ye X,Wu J,Zhou J,Wang F,Chen Q,Fu L,Li S,Huang Y,Cui Zdoi
10.1016/j.pep.2015.06.009subject
Has Abstractpub_date
2017-01-01 00:00:00pages
122-127eissn
1046-5928issn
1096-0279pii
S1046-5928(15)00148-5journal_volume
129pub_type
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