Extracellular Production and Purification of the ?-glucanase in Pichia pastoris Expression System
نویسندگان
چکیده
In this study, Rhizomucor miehei ?-1,3-1,4-glucanase gene was expressed under the regulation of AOX1 promoter in Pichia pastoris expression system, and clone providing highest production level determined. The codon-optimized ligated into vector pPICZ?A transferred competent P. KM71H cells. Thirty transformants were selected from plates containing different concentrations zeocin cultured test tubes for protein production. Glucanase enzyme activities supernatant samples measured ten clones showing glucanase activity proteins also analyzed by SDS-PAGE observed 2 bands, approximately 34 38 kDa. Protein performed 72 hours 200 mL induction medium (BMMY) with recombinant purified Ni-NTA affinity chromatography. After purification, it determined that reached 796 mg/L level. analysis each step purification procedure showed two bands represent enzyme.
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ژورنال
عنوان ژورنال: Erzincan University Journal of Science and Technology
سال: 2021
ISSN: ['1307-9085', '2149-4584']
DOI: https://doi.org/10.18185/erzifbed.972627