Cloning of Laccase Gene from Coriolus Versicolor and Optimization of Culture Conditions for Lcc1 Expression in Pichia Pastoris
A laccase cDNA lcc1 (GenBank accession number HM137002), without native signal peptide, was cloned by RT-PCR from total RNA of Coriolus versicolor. Recombination expression vector pPICZαA-lcc1 was constructed and transformed into Pichia pastoris KM71H after lineared. Recombination laccase was expressed at a higher level. Single factors of fermentation conditions of Pichia pastoris KM71H for laccase production were optimized. The results showed optimal culture conditions were as follows: medium initial pH 7.5, Cu2+ concentration 0.5mmol/L, methanol additive amount 1.0% and shaker rotate speed 210r/min. Furthermore, induction at low temperature was more suitable for lcc1 secretion. And addition of appropriate amount peptone and tyrosine in culture medium could enhanced lcc1 yields and reduce its degradation.
Zhong Cao, Yinghe He, Lixian Sun and Xueqiang Cao
L. Y. He et al., "Cloning of Laccase Gene from Coriolus Versicolor and Optimization of Culture Conditions for Lcc1 Expression in Pichia Pastoris", Advanced Materials Research, Vols. 236-238, pp. 1039-1044, 2011