Document Detail


Production of bovine-pancreatic-trypsin-inhibitor homologues in Escherichia coli and their characterization.
MedLine Citation:
PMID:  8573289     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Biologically active bovine pancreatic trypsin inhibitor (BPTI) was produced in Escherichia coli using an OmpA leader-peptide fusion-protein system, and BPTI homologues were generated by cassette mutagenesis. Amino acids in the reactive loop of alpha 1-proteinase inhibitor (alpha 1-PI) were incorporated into the reactive loop of BPTI in a stepwise approach such that the contribution of individual amino acids could be assessed. The introduction of mutations into BPTI diminished the yield of heterologous protein relative to wild-type BPTI. However, for three BPTI homologues sufficient material was isolated to allow characterization of the proteins by electrospray MS and N-terminal peptide sequencing.
Authors:
J A Chesshyre; J A Kraunsoe; G Lowe
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and applied biochemistry     Volume:  22 ( Pt 3)     ISSN:  0885-4513     ISO Abbreviation:  Biotechnol. Appl. Biochem.     Publication Date:  1995 Dec 
Date Detail:
Created Date:  1996-03-11     Completed Date:  1996-03-11     Revised Date:  2007-03-21    
Medline Journal Info:
Nlm Unique ID:  8609465     Medline TA:  Biotechnol Appl Biochem     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  269-80     Citation Subset:  IM    
Affiliation:
Dyson Perrins Laboratory, University of Oxford, U.K.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / metabolism
Animals
Bacterial Outer Membrane Proteins / genetics,  metabolism
Base Sequence
Cattle
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
DNA / chemistry,  metabolism
Electrophoresis, Polyacrylamide Gel
Escherichia coli / metabolism*
Escherichia coli Proteins*
Fermentation
Mass Spectrometry
Molecular Sequence Data
Molecular Weight
Mutagenesis, Insertional
Mutagenesis, Site-Directed
Plasmids / metabolism
Protein Precursors / genetics,  metabolism
Trypsin Inhibitor, Kazal Pancreatic / biosynthesis*,  chemistry,  genetics,  metabolism
alpha 1-Antitrypsin / metabolism
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Bacterial Outer Membrane Proteins; 0/Escherichia coli Proteins; 0/Protein Precursors; 0/alpha 1-Antitrypsin; 0/outer membrane protein A precursor (E coli); 50936-63-5/Trypsin Inhibitor, Kazal Pancreatic; 9007-49-2/DNA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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