High-level Expression and Secretion of Serratia marcescens Metalloprotease Inhibitor in Bacillus subtilit by Aid of Subtilisin Promotor and Signal Sequence

다국어 초록 (Multilingual Abstract)

Metalloprotease inhibitor(SmaPI) of a gram-negetive bacterium, Serratia marcescens, was expressed and secreted in a heterologous host, bacillus subtilis DB431, by aid of a subtillisin promotor and signal sequence. The DNA fragment containing the coding sequence for SmaPI was amplified by polymerase chain reaction, and the amplified-DNA product was inserted into the downstream region of a subtillisin signal sequence. The recombinant SmaPI expressed in bacillus subtilis DB431 was sereted into the culture medium in a large amount. After cultivation for 32 h, the amount of SmaPI secreted into the culture medium reached about 100㎎/liter when estimated by measuring inhibitory activities toward Serratia marcescens metalloprotease(SMP). The NH_2terminal amino acid sequencing analysis confirmed that authentic SmaPI and the recombinant SmaPI have the same NH_2 -terminal amino acid sequences. The inhibitory activity of the purified recombinant SmaPI was found to be nearly equivalent to that of authentic SmaPI.