A Comparative Study of Recombinant Expression of Three Aminopeptidases in Escherichia coli

doi:10.7506/spkx1002-6630-201323042

Abstract

Abstract:

The genes encoding aminopeptidase GSAP, BSAP and BLAP were respectively amplified from Geobacillus
stearothermophilus IFO12589, Bacillus subtilis 168 and Bacillus licheniformis 14580. These amplified DNA fragments
were individually inserted into the expression vector E. coli pET-28a to construct plasmids pET28a-BLAP, pET28a-BSAP
and pET28a-GSAP, respectively. The expression of these genes was confirmed by activity analysis, and then the expressed
enzymes were characterized. The activities of both recombinant BSAP and GSAP were 1500 U/L, which were higher than
that of BLAP. The optimum temperatures were 50, 75 ℃ and 60 ℃ for BSAP, GSAP and BLAP, respectively, and the
optimum pH values were all 9.0. They were stable at pH 8.5—9.0. BSAP was obviously activated by 0.l mmol/L Co2+,
reaching a maximum of 195.6% of its original activity, but inhibited to different extents by other divalent metal ions, with
Zn2+ being the strongest inhibitor. In addition, the recombinant plasmids pET28a-BSAP and pET28a-GSAP were more stable
than pET28a-BLAP in E. coli.

Key words: aminopeptidase, recombinant expression, enzymatic reaction

CLC Number:

Q786

ZHANG Jie1，ZHANG Liang1，HUANG Wu-ning2，SHI Gui-yang1,*. A Comparative Study of Recombinant Expression of Three Aminopeptidases in Escherichia coli[J]. FOOD SCIENCE, 2013, 34(23): 200-205.

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