DIRECT EXPRESSION OF PEPTIDE INTO CULTURE MEDIUM

PROBLEM TO BE SOLVED: To produce peptide in high yield with an efficient expression vector, and to provide high yield culturing techniques and other improvements which permit high yield recovery of excreted peptide of interest from the culture media, without overly disrupting the integrity of a cell...

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Bibliographische Detailangaben
Hauptverfasser:	MEENAN CHRISTOPHER P, RAY MARTHA V L, MEHTA NOZAR M, CONSALVO ANGELO P
Format:	Patent
Sprache:	eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS THEREOF CULTURE MEDIA ENZYMOLOGY FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE HUMAN NECESSITIES HYGIENE MEDICAL OR VETERINARY SCIENCE METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING ORGANIC CHEMISTRY PEPTIDES PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE
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Beschreibung
Zusammenfassung:	PROBLEM TO BE SOLVED: To produce peptide in high yield with an efficient expression vector, and to provide high yield culturing techniques and other improvements which permit high yield recovery of excreted peptide of interest from the culture media, without overly disrupting the integrity of a cell membrane.SOLUTION: The expression vector includes two transcription cassettes in tandem, each cassette comprising: (a) a coding region with nucleic acids encoding a peptide product coupled in the reading frame 3' of nucleic acids encoding a signal peptide selected from OmpA, pelB, OmpC, OmpF, OmpT, β-la, PhoA, PhoS and StaphA; and (b) a control region linked operably with the coding region, where the control region comprises two promoters in tandem, that are a lac promoter and a tac promoter existing upstream of the lac promoter, and at least one ribosome binding site. The expression vector further includes at least one secretion enhancing peptide selected from SecE, SecY, and prlA-4.