公开(公告)号：US5856166A

公开(公告)日：1999-01-05

申请号：US265310

申请日：1994-06-24

申请人： Daniel Bartfeld ,  Michael J. Butler ,  Dany Hadary ,  David L. Jenish ,  Timothy J. Krieger ,  Lawrence T. Malek ,  Gisela Soostmeyer ,  Eva Walczyk ,  Phyllis Krygsman ,  Sheila Garven

发明人： Daniel Bartfeld ,  Michael J. Butler ,  Dany Hadary ,  David L. Jenish ,  Timothy J. Krieger ,  Lawrence T. Malek ,  Gisela Soostmeyer ,  Eva Walczyk ,  Phyllis Krygsman ,  Sheila Garven

IPC分类号： C07K14/475 ,  C07K14/505 ,  C07K14/53 ,  C07K14/535 ,  C07K14/54 ,  C12N9/48 ,  C12N9/52 ,  C12N15/57 ,  C12N15/62 ,  C12N15/76 ,  C12N1/20

CPC分类号： C12N15/76 ,  C07K14/475 ,  C07K14/505 ,  C07K14/53 ,  C07K14/535 ,  C07K14/5418 ,  C12N15/62 ,  C12N15/625 ,  C12N9/48 ,  C12N9/52 ,  C07K2319/00 ,  C07K2319/02

摘要： A family of proteases endogenous to Streptomyces cells degrades heterologous proteins secreted from Streptomyces host cells. The previously unidentified proteases include (1) tripeptidyl aminopeptidase--Streptomyces ("Tap"), (2) a Streptomyces protease ("Ssp") which displayed significant amino acid sequence homology to Subtilisin BPN' and showed an ability to remove tripeptides from the amino termini of proteins and peptides, and (3) other proteases derived from Streptomyces which degraded certain substrates under certain conditions. Degradation was alleviated by selective inhibition of secreted proteases or by using hosts with impaired capabilities to produce proteases. An irreversible inhibitor was designed based upon the mechanism and substrate specificity of the target protease. Hosts secreting high amounts of proteases were selected. Impaired hosts were produced by deleting or altering the nucleotide sequence for the proteases.

摘要翻译： 链霉菌细胞内源的蛋白酶家族降解从链霉菌宿主细胞分泌的异源蛋白质。 以前不明确的蛋白酶包括（1）三肽基氨基肽酶 - 链霉菌（“Tap”），（2）与枯草杆菌蛋白酶BPN'显示显着的氨基酸序列同源性的链霉菌蛋白酶（“Ssp”），并显示出从氨基 蛋白质和肽的末端，和（3）衍生自链霉菌的其它蛋白酶，其在某些条件下降解某些底物。 通过选择性抑制分泌的蛋白酶或通过使用能力下降的宿主产生蛋白酶来减轻降解。 基于靶蛋白酶的机制和底物特异性设计不可逆抑制剂。 选择分泌大量蛋白酶的宿主。 通过缺失或改变蛋白酶的核苷酸序列产生受损宿主。

公开(公告)号：US6127144A

公开(公告)日：2000-10-03

申请号：US951742

申请日：1997-10-16

申请人： Daniel Bartfeld ,  Michael J. Butler ,  Dany Hadary ,  David L. Jenish ,  Timothy J. Krieger ,  Lawrence T. Malek ,  Gisela Soostmeyer ,  Eva Walczyk ,  Phyllis Krygsman ,  Sheila Garven

发明人： Daniel Bartfeld ,  Michael J. Butler ,  Dany Hadary ,  David L. Jenish ,  Timothy J. Krieger ,  Lawrence T. Malek ,  Gisela Soostmeyer ,  Eva Walczyk ,  Phyllis Krygsman ,  Sheila Garven

IPC分类号： C12N9/52 ,  C12P21/06 ,  A61K38/06 ,  C12N1/12 ,  C12N9/48

CPC分类号： C12N9/52 ,  C07K2319/02

摘要： An aminopeptidase inhibitor is used when expressing heterologous protein in a bacterial host, such as Streptomyces. Use of such an inhibitor inhibits degradation of the heterologous protein by aminopeptidases. Inhibitors are designed based upon the mechanism and substrate specificity of the target protease and expressed protein.

摘要翻译： 当在细菌宿主例如链霉菌中表达异源蛋白质时，使用氨肽酶抑制剂。 使用这种抑制剂抑制氨基肽酶对异源蛋白质的降解。 基于靶蛋白酶和表达蛋白质的机制和底物特异性设计抑制剂。

公开(公告)号：US5616485A

公开(公告)日：1997-04-01

申请号：US173508

申请日：1993-12-23

申请人： Dany Hadary ,  Daniel Bartfeld ,  Michael J. Butler ,  David Jenish ,  Timothy Krieger ,  Lawrence T. Malek ,  Gisela Soostmeyer ,  Eva Walcyzk

发明人： Dany Hadary ,  Daniel Bartfeld ,  Michael J. Butler ,  David Jenish ,  Timothy Krieger ,  Lawrence T. Malek ,  Gisela Soostmeyer ,  Eva Walcyzk

IPC分类号： C07K14/475 ,  C07K14/505 ,  C07K14/53 ,  C07K14/535 ,  C07K14/54 ,  C12N9/48 ,  C12N9/52 ,  C12N15/57 ,  C12N15/62 ,  C12N15/76 ,  C12N15/31

CPC分类号： C12N15/76 ,  C07K14/475 ,  C07K14/505 ,  C07K14/53 ,  C07K14/535 ,  C07K14/5418 ,  C12N15/62 ,  C12N15/625 ,  C12N9/48 ,  C12N9/52 ,  C07K2319/00 ,  C07K2319/02

摘要： A family of proteases endogenous to Streptomyces cells degrade heterologous proteins secreted from Streptomyces host cells. The previously unidentified proteases include (1) tripeptidyl aminopeptidase--Streptomyces ("Tap"), (2) a Streptomyces protease ("Ssp") which displayed significant amino acid sequence homology to Subtilisin BPN' and showed an ability to remove tripeprides from the amino termini of proteins and peptides, and (3) other proteases derived from Streptomyces which degraded certain substrates under certain conditions. Degradation was alleviated by selective inhibition of secreted proteases or by using hosts with impaired capabilities to produce proteases. An irreversible inhibitor was designed based upon the mechanism and substrate specificity of the target protease. Hosts secreting high amounts of proteases were selected. Impaired hosts were produced by deleting or altering the nucleotide sequence for the proteases.

摘要翻译： 链霉菌细胞内源的蛋白酶家族降解从链霉菌宿主细胞分泌的异源蛋白质。 以前不明确的蛋白酶包括（1）三肽基氨基肽酶 - 链霉菌（“Tap”），（2）与枯草杆菌蛋白酶BPN'显示显着氨基酸序列同源性的链霉菌蛋白酶（“Ssp”），并且显示出从氨基 蛋白质和肽的末端，和（3）衍生自链霉菌的其它蛋白酶，其在某些条件下降解某些底物。 通过选择性抑制分泌的蛋白酶或通过使用能力下降的宿主产生蛋白酶来减轻降解。 基于靶蛋白酶的机制和底物特异性设计不可逆抑制剂。 选择分泌大量蛋白酶的宿主。 通过缺失或改变蛋白酶的核苷酸序列产生受损宿主。