Efficient secretory expression of an alkaline pectate lyase gene from Bacillus subtilis in E. coli and the purification and characterization of the protein
作者： Bin Zhuge , Guo-Cheng Du , Wei Shen , Jian Zhuge , Jian Chen
DOI: 10.1007/S10529-006-9249-6
关键词:
摘要: The gene encoding pectate lyase (PL) from Bacillus subtilis WSHB04-02 was amplified by PCR, fused with a periplasmic secretion signal peptide sequence, pelB, pET22b(+), cloned and expressed in Escherichia coli cells using temperature control vector, pHsh. recombinant E. coil grown 5 l fermentor. PL secreted broth at 22 U l−1 after 20 h when increased 30°C to 42°C. enzyme purified homogeneity as judged SDS-PAGE. It optimally active pH 9.4 50°C over 30 min. Analysis of polygalacturonic acid (PGA) degradation products electrospray ionization (ESI)-mass spectrometry (MS) indicated that produced mixture unsaturated oligo-galacturonides including tri-galacturonic bi-galacturonic but not mono-galacturonic acid.
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