Secretory expression in Escherichia coli and single-step purification of a heat-stable alkaline protease
作者： Zhibiao Fu , Suhali Bt Ab Hamid , Che Nyonya A Razak , Mahiran Basri , Abu Bakar Salleh
DOI: 10.1016/S1046-5928(02)00637-X
关键词:
摘要: Abstract Bacteriocin release proteins (BRPs) can be used for the of heterologous from Escherichia coli cytoplasm into culture medium. The gene a highly thermostable alkaline protease was cloned Bacillus stearothermophilus F1 by polymerase chain reaction. recombinant efficiently excreted medium using E. XL1-Blue harboring two vectors: pTrcHis bearing and pJL3 containing BRPs. Both vectors contain lac promoter–operator system. In presence 40 μM IPTG, BRP were expressed mature released This opens way large-scale production this in coli. enzyme purified through one-step heat treatment at 70 °C 3 h method to near homogeneity. showed pH optimum 9.0, temperature 80 °C, stable 24 h range 8.0 10.0. exhibited high degree thermostability with half-life 4 h 85 °C, 25 min 90 °C, inhibited serine inhibitor phenylmethylsulfonyl fluoride (PMSF).
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