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Heterologous production of a temperature and pH-stable laccase from Bacillus vallismortis fmb-103 in Escherichia coli and its application.

Process Biochemistry.. 2017-02; 
Jianna Sun, Meixia Zheng, Zhaoxin Lu, Fengxia Lu, Chong Zhang. Laboratory of Enzyme Engineering, College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, PR China.
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摘要

To enhance laccase yield, the laccase gene from Bacillus vallismortis fmb-103 was cloned and heterologously expressed in Escherichia coli BL21 (DE3) cells. The auto-induction strategy was applied during fermentation, and the process was controlled, as follows: Cu2+ was added when the optical density at 600 nm (OD600) was 0.3, the fermentation temperature was adjusted to 16 °C when the OD600 was 0.9, and fermentation was stopped after 50 h. The yield of recombinant laccase was up to 3420 U/L, as assayed by 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid). Recombinant laccase was purified 4.47-fold by heating for 10 min at 70 °C and dialyzing against 50-60% ammonium sulfate, retained more than 50% ac... More

关键词

Bacillus vallismortis fmb-103; Laccase; Heterologous production; Characterization; Malachite green degradation