(The) expression of streptomyces griseus protease and streptokinase genes and role of the propeptide = 스트렙토마이세스 그리세우스 프로테아제와 스트렙토키나제 유전자의 발현 및 프로펩타이드의 역할

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Streptomyces griseus protease A (SGPA), B (SGPB) and Trypsin (SGT) are the extracellular serine proteases from Streptomyces griseus. SGPA and SGPB are chymotrypsin-like protease and SGT is a trypsin-like protease. It is such an interesting feature that prepro-sequence of SGPA and SGPB quite differ from that of SGT. SGPA, SGPB and SGT are very homologous not only to one another, but also to mammalian serine proteases. Streptokinase (SK) is an extracellular protein from several strains of streptococci that functions a plasminogen activator, a fibrinolytic agent without its protease activity. It has been proposed that these serine family proteases were related to Streptokinase in amino acid homology and molecular evolution. In our previous report, the genes encoding S. griseus protease A (sprA), B (sprB), Trypsin (sprT) were cloned from S. griseus ATCC 10137. In this study, the nucleotide sequence of SGPA and SGPB genes was determined and characterized. And several vectors for expression and secretion were constructed to develop the appropriate expression system, and the wild and mutated types of SGPA, SGPB, SGT and SK genes were expressed in E. coli, Bacillus subtilis and Streptomyces lividans. In addition the role of the propeptide of the microbial serine proteases was investigated. The amino acid sequence deduced from the nucleotide sequence suggests that S. griseus protease A and B are produced as a precursor consisting of three portions. In the case of SGPB, an amino-terminal pre-sequence (38 amino acid residues), a pro-sequence (76 amino acid residues), and mature protease (185 amino acid residues). The first expression vectors for sprA and sprB were constructed by subcloning the gene fragments into multi-cloning site of E. coli-S. lividans shuttle vector. S. lividans 1326 transformed with the recombinant plasmids showed clear zone activity around its colony on the skim milk-YD plate. When the protease activity from culture medium was assayed using artifici...
Advisors
Byun, Si-Myung변시명
Description
한국과학기술원 : 생물과학과,
Publisher
한국과학기술원
Issue Date
1998
Identifier
135081/325007 / 000905802
Language
eng
Description

학위논문(박사) - 한국과학기술원 : 생물과학과, 1998.2, [ xii, 130 p. ]

Keywords

Propeptide; Expression folding; Streptomyces griseus protease; Streptokinase; 폴딩; 발현; 프로펩타이드; 스트렙토키나제; 스트렙토미세스 그리세우스 프로테아제

URI
http://hdl.handle.net/10203/27418
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=135081&flag=dissertation
Appears in Collection
BS-Theses_Ph.D.(박사논문)
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