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Proteins, Expression, Isolation and Analysis> | … fragments were purified with the AxyPrep DNA purification kit (Axygen) and cloned into pET24a with BamHI and HindIII sites to construct the plasmid pET-M6A, which was further transformed into the competent cells of E. coli …(genscript) | Get A Quote |
The gene M6A coding a novel cold-adapt endoglucanase was cloned from Microbacterium kitamiense Sa12 isolated from a wasteland in Saga, Qinghai-Tibetan Plateau. The deduced protein sequence encoded a 411-residue polypeptide sharing similar identities with glycosyl hydrolase family 6 enzymes. The recombinant M6A displayed maximum hydrolysis activity of 1.51 U/mg toward soluble cellulose substrate, CMC at 35°C and pH 5.0, and the Km and Vmax value were 2.12 mg/mL and 15.33 µmol/min*mg, respectively. Interestingly, M6A exhibited significant activity even at ice cold condition, showing 30–40% relative activity at 0–5°C, and had good tolerance to Li+, K+, NH4+, Rb+, Ca2+, Mg2+ Zn2+, Mn2+, and Ni2+, expect diva... More