Method of producing microbial transglutaminase
The present invention provides a neutral metalloprotease from actinomycetes which selectively cleaves a pro-structure part of a microbial protransglutaminase and a gene encoding said neutral metalloprotease. An active microbial transglutaminase having the pro-structure part cleaved can be obtained by culturing a microorganism into which a gene encoding the neutral metalloprotease from actinomycetes according to the present invention has been introduced, where by producing the neutral metalloprotease from actinomycetes, and reacting it on a microbial protransglutaminase.
1. A biochemical method of producing an active Streptoverticillium transglutaminase from a Streptoverticillium protransglutaminase comprising:
A) contacting a neutral Streptoverticillium mobaraense IFO 13819 metalloprotease with the protransglutaminase, and
B) recovering the active Streptoverticillium microbial transglutaminase;
wherein said metalloprotease comprises:
a molecular weight of about 71,000,
an optimum pH of 7.0,
an optimum temperature of about 55° C.,
an activity to cleave the peptide of SEQ ID NO: 11; and
wherein said metalloprotease is strongly inhibited by ethylene diamine tetraacetic acid, 1,10-phenanthroline, phosphoramidon, dithiothreitol, and Streptomyces subtilisin inhibitor (SSI) from actinomycetes.
2. The method according to claim 1 , wherein said microorganism is a coryneform bacterium.
3. The method according to claim 1 , wherein the metalloprotease comprises, as an N-terminal amino acid sequence, the sequence of SEQ ID NO: 2.