Protein Expression and Purification, Vol.146, 23-26, 2018
Kinetic properties analysis of beta-mannanase from Klebsiella oxytoca KUB-CW2-3 expressed in Escherichia coli
Endo-1,4-beta-mannanase is an enzyme that can catalyze the random hydrolysis of beta-1,4-mannosidic linkages in the main chain of mannans, glucomannans and galactomannans and offers many applications in different biotechnology industries. Purification and kinetic properties of the endo-1,4-beta-mannanase from recombinant Escherichia colt strain KMAN-3 were examined. Recombinant beta-mannanase (KMAN-3) was purified 50.5 fold using Ni-NTA Agarose resin and specific activity of 11900 U/mg protein was obtained. Purified KMAN-3 showed a single band on SDS-PAGE with a molecular weight of 43 kDa. K-m and V-max values of KMAN-3 on ivory nut mannan, locust bean gum, defatted copra meal and konjac glucomannan were 243, 3.83 x 10(5) 37 and 2.13 x 10(6) mg ml(-1) and 2940, 61,100, 3930 and 1.56 x 10(10) mg(-1), respectively. Carboxymethyl cellulose was not digested by KMAN-3.