Gene Cloning, Expression, and Characterization of an Exo-inulinase from Paenibacillus polymyxa ZJ-9.

Gene Cloning, Expression, and Characterization of an Exo-inulinase from Paenibacillus polymyxa ZJ-9. Appl Biochem Biotechnol. 2014 May 8; Authors: Gao J, Xu YY, Yang HM, Xu H, Xue F, Li S, Feng XH Abstract An inulinase-producing strain, Paenibacillus polymyxa ZJ-9, was isolated from natural sources to produce R,R-2,3-butanediol via one-step fermentation of raw inulin extracted from Jerusalem artichoke tubers. The inulinase gene from P. polymyxa ZJ-9 was cloned and overexpressed in Escherichia coli BL21 (DE3), and the purified recombinant inulinase was estimated to be approximately 56 kDa by both sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and gel filtration chromatography. This result suggests that the active form of the inulinase is probably a monomer. Terminal hydrolysis fructose units from the inulin indicate that enzymes are exo-inulinase. The purified recombinant enzyme showed maximum activity at 25 °C and pH 6.0, which indicate its extreme suitability for industrial applications. Zn(2+), Fe(2+), and Mg(2+) stimulated the activity of the purified enzyme, whereas Co(2+), Cu(2+), and Ni(2+) inhibited enzyme activity. The K m and V max values for inulin hydrolysis were 1.72 mM and 21.69 μmol min(-1) mg(-1) protein, respectively. The same parameters toward sucrose were 41.09 mM and 78.7 μmol min(-1) mg(-1) protein, respectively. Considering its substrate specificity and other enzymatic characte...
Source: Applied Biochemistry and Biotechnology - Category: Biochemistry Authors: Tags: Appl Biochem Biotechnol Source Type: research