Protein disulfide-isomerase and production thereof
AssigneeTakeda Chemical Industries, Ltd.
InventorsKumao Toyoshima, Ryuya Horiuchi, Kiyoshi Yamauchi, Tadashi Yamamoto, Koichi Igarashi
Life of this patent
Estimating the term…
Abstract
Protein disulfide isomerase, which catalyzes the exchange reaction between sulfhydryl and disulfide in a protein for formation of the most stable natural type disulfide bonds, is useful for formation of natural type disulfide bonds in a protein which is produced in a prokaryotic cell.
Claims24
An isolated DNA encoding the amino acid sequence of FIG. 3.
An isolated DNA encoding the amino acid sequence:
- ______________________________________ MetAlaProGluGluGluAspHisValLeuValLeuArgLysSerAsnPheAlaGluAla LeuAlaAlaHisLysTyrLeuLeuValGluPheTyrAlaProTrpCysGlyHisCysLys AlaLeuAlaProGluTyrAlaLysAlaAlaGlyLysLeuLysAlaGluGlySerGluIle ArgLeuAlaLysValAspAlaThrGluGluSerAspLeuAlaGlnGlnTyrGlyValArg GlyTyrProThrIleLysPhePheArgAsnGlyAspThrAlaSerProLysGluTyrThr AlaGlyArgGluAlaAspAspIleValAsnTrpLeuLysLysArgThrGlyProAlaAla ThrThrLeuProAspGlyAlaAlaAlaGluSerLeuValGluSerSerGluValAlaVal IleGlyPhePheLysAspValGluSerAspSerAlaLysGlnPheLeuGlnAlaAlaGlu AlaIleAspAspIleProPheGlyIleThrSerAsnSerAspValPheSerLysTyrGln LeuAspLysAspGlyValValLeuPheLysLysPheAspGluGlyArgAsnAsnPheGlu GlyGluValThrLysGluAsnLeuLeuAspPheIleLysHisAsnGlnLeuProLeuVal IleGluPheThrGluGlnThrAlaProLysIlePheGlyGlyGluIleLysThrHisIle LeuLeuPheLeuProLysSerValSerAspTyrAspGlyLysLeuSerAsnPheLysThr AlaAlaGluSerPheLysGlyLysIleLeuPheIlePheIleAspSerAspHisThrAsp AsnGlnArgIleLeuGluPhePheGlyLeuLysLysGluGluCysProAlaValArgLeu IleThrLeuGluGluGluMetThrLysTyrLysProGluSerGluGluLeuThrAlaGlu ArgIleThrGluPheCysHisArgPheLeuGluGlyLysIleLysProHisLeuMetSer GlnGluLeuProGluAspTrpAspLysGlnProValLysValLeuValGlyLysAsnPhe GluAspValAlaPheAspGluLysLysAsnValPheValGluPheTyrAlaProTrpCys GlyHisCysLysGlnLeuAlaProIleTrpAspLysLeuGlyGluThrTyrLysAspHis GluAsnIleValIleAlaLysMetAspSerThrAlaAsnGluValGluAlaValLysVal HisSerPheProThrLeuLysPhePheProAlaSerAlaAspArgThrValIleAspTyr AsnGlyGluArgThrLeuAspGlyPheLysLysPheLeuGluSerGlyGlyGlnAspGly AlaGlyAspAspAspAspLeuGluAspLeuGluGluAlaGluGluProAspMetGluGlu AspAspAspGlnLysAlaValLysAspGluLeu. ______________________________________
A recombinant vector comprising a nucleotide sequence which encodes a polypeptide comprising the amino acid sequence of FIG. 3 and which is operably linked to a promoter sequence, wherein said recombinant vector replicates and expresses said polypeptide when transformed into in a microorganism.
transformed microorganism comprising a recombinant vector, wherein said recombinant vector comprises a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of FIG. 3, which nucleotide sequence is operably linked to a promoter sequence, and wherein said recombinant vector replicates and expresses said polypeptide in said transformed microorganism.
A method for producing a polypeptide comprising the amino acid sequence of FIG. 3, which method comprises the steps of:
- (i) cultivating a transformed microorganism comprising a recombinant vector, wherein said recombinant vector comprises a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of FIG. 3, which nucleotide sequence is operably linked to a promoter sequence, and wherein said recombinant vector replicates and expresses said polypeptide in said transformed microorganism;
- (ii) producing and accumulating said polypeptide intracellularly in said transformed microorganism;
- and (iii) recovering said polypeptide from said transformed microorganism.
A method of producing a transformed microorganism comprising a recombinant vector, wherein said recombinant vector comprises a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of FIG. 3, which nucleotide sequence is operably linked to a promoter sequence, and wherein said recombinant vector replicates and expresses said polypeptide in said transformed microorganism;
- which method comprises the steps of:
- (i) transforming a microorganism with said recombinant vector, and (ii) recovering said transformed microorganism.
A method for producing a recombinant vector, wherein said recombinant vector comprises a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of FIG. 3, which nucleotide sequence is operably linked to a promoter sequence, and wherein said recombinant vector replicates and expresses said polypeptide in a microorganism;
- which method comprises the steps of:
- (i) inserting an isolated DNA comprising said nucleic acid sequence into a site down stream from a promoter sequence in a vector that replicates in said microorganism so that said nucleic acid sequence is operably linked to said promoter sequence, and (ii) recovering said recombinant vector.
A method for expressing a biologically active polypeptide, wherein said biologically active polypeptide comprises the amino acid sequence of FIG. 3, which method comprises the step of:
- cultivating a transformed animal cell or a transformed microorganism belonging to genus Escherichia or genus Saccharomyces, wherein said transformed animal cell or said transformed microorganism is transformed with a recombinant vector, wherein said recombinant vector comprises a nucleotide sequence encoding said biologically active polypeptide, which nucleotide sequence is operably linked to a promoter sequence, and wherein said recombinant vector replicates and expresses said biologically active polypeptide in said animal cell or microorganism.
Dependencies are inferred from the claim text.