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Cloning of a 6.6-kb BamHI digested chromosomalDNA from S. griseus IFO1350 revealed the presence of anadditional gene encoding a novel trypsin-like enzyme, namedSprU. The SprU protein shows a high homology (79%been reported as a bacterial trypsin in the same strain. Theamino acid sequence deduced from the nucleotide sequenceof the sprU gene suggests that SprU is produced as aprecursor consisting of an amino-terminal presequence (29amino acid residues), prosequence (4 residues), and maturetrypsin consisting of 222 amino acids with a molecular weightof 22.94 kDa and a calculated pI of 4.13. The serine, histidine,and aspartic acid residues composing the catalytic triad oftypical serine proteases are also wel conserved. When theby the enzymatic hydrolysis of the artificial chromogenicsubstrate, N-α-benzoyl-DL-arginine-ρ-nitroanilide, the S. lividanstransformant with pWHM3-U gave 3 times higher activitythan that of control. When the same recombinant plasmidwas introduced into S. griseus, however, the gene dosageeffect was not so significant, as in the cases of other genesencoding serine proteases, such as sprA, sprB, and sprD.Although two trypsins, SprU and SGT, have a high degree ofhomology, the pI values, the gene dosage efect in S. griseus,and the gene arrangement adjacent to the two genes arevery diferent, suggesting that the biochemical and biologicalfunction of the SprU might be quite diferent from that ofthe SGT.

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