Abstract: 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.

Abstract: By culturing a methanol-assimilating bacterium which harbors a DNA construct which contains a promoter sequence that functions in the methanol-assimilating bacterium and a nucleotide sequence that encodes a polypeptide containing a signal sequence and an objective protein which is functionally connected to the promoter sequence, in a liquid medium containing methanol as a major carbon source, the bacterium is allowed to secrete the objective protein, and the secreted objective protein is recovered.

Abstract: The present invention provides a method for efficiently producing an industrially useful protein in coryneform bacteria, and more particularly, a method for efficiently producing a protein for which secretion was difficult using conventional protein secretion pathways. In particular, the present invention provides a method for efficiently producing heterologous proteins comprising culturing coryneform bacteria containing an genetic construction containing a promoter sequence which functions in coryneform bacteria, a nucleic acid sequence encoding a Tat system-dependent signal peptide region, and a nucleic acid sequence encoding a heterologous protein, in the direction from 5?-end to 3?-end, and secretory producing the heterologous protein by coryneform bacteria.

Abstract: The object of the present invention is to provide a method of producing a heterologous protein by making a coryneform bacterium to produce and efficiently extracellularly secrete (secreto-production) an industrially useful heterologous protein. According to the present invention, a genetic construct is used where a gene sequence encoding an intended protein which is ligated to the downstream of a sequence encoding the signal peptide derived from a coryneform bacterium, the gene construct is introduced into a mutant coryneform bacterium which has a capacity of secreting the heterologous protein at least 2-fold higher than the wild type Corynebacterium glutamicum ATCC 13869, the mutant coryneform bacterium is cultured and the extracellularly released heterologous protein is recovered.

Abstract: 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.

Abstract: In gene expression frequency analysis, a vector primer to which each cDNA is ligated is formed by synthesizing the cDNA from each mRNA using by a vector primer having a poly(T) sequence; each cDNA sequence is converted to a tag on the vector primer; a concatemer is formed by ligating the obtained tags via a sequence that enables recognition of ends of the tags; and the nucleotide sequence of the concatemer is analyzed.

Abstract: The object of the present invention is to provide a method of producing a heterologous protein by making a coryneform bacterium to produce and efficiently extracellularly secrete (secreto-production) an industrially useful heterologous protein. According to the present invention, a genetic construct is used where a gene sequence encoding an intended protein which is ligated to the downstream of a sequence encoding the signal peptide derived from a coryneform bacterium, the gene construct is introduced into a mutant coryneform bacterium which has a capacity of secreting the heterologous protein at least 2-fold higher than the wild type Corynebacterium glutamicum ATCC 13869, the mutant coryneform bacterium is cultured and the extracellularly released heterologous protein is recovered.

Abstract: The present invention relates to a process for secretory production of a foreign protein, in particular, transglutaminase by a coryneform bacterium.

Abstract: There are disclosed (i) a purification process for obtaining a human BCDF having the intramolecular disulfide linkage and the stereostructure of natural type human BCDF which comprises subjecting to an oxidation reaction and a refolding treatment a reduced type human BCDF obtained by culturing a microorganism having a human BCDF gene integrated therein and solubilized with guanidine hydrochloride, characterized in that after the oxidation reaction, a gel filtration chromatographic treatment is conducted under the conditions of the guanidine hydrochloride concentration adjusted to 4-7M; (ii) a purification process for obtaining a natural type human BCDF monomer by removing the organic solvent from an organic solvent-containing solution of human BCDF, characterized in that the solution is passed through a gel filtration chromatographic column equilibrated with an organic solvent, followed by eluting according to a stepwise or linear gradient program; and (iii) a human BCDF purification process comprising an ion