Claims for Patent: 7,897,366

Summary for Patent: 7,897,366

Title:	Method for simultaneous display of target protein and cofactor regenerating enzyme on cell surface
Abstract:	The present invention relates to a method for simultaneously surface expressing a target protein using a cofactor and an enzyme regenerating the cofactor on the cell surface. According to the present invention, it is possible to provide a microorganism capable of simultaneously surface expressing a target protein using a cofactor to transform a biochemical material at a high efficiency and an enzyme generating the cofactor without adding an expensive cofactor in a large amount.
Inventor(s):	Lee; Sang Yup (Daejeon, KR), Lee; Seung-Hwan (Daejeon, KR)
Assignee:	Korea Advanced Institute of Science and Technology (Daejeon, KR)
Application Number:	11/740,390
Patent Claims:	1. A method for simultaneously expressing a cofactor regenerating enzyme and a target protein using the cofactor on the cell surface, the method comprising the steps of: (a) preparing a recombinant vector for regenerating the cofactor comprising DNA having a gene encoding an enzyme for regenerating the cofactor linked to a gene encoding outer membrane protein C (OmpC) so that the enzyme regenerating the cofactor is expressed in the form fused with OmpC; (b) preparing a recombinant vector for expressing the target protein comprising DNA having a gene encoding the target protein linked to a gene encoding OmpC so that the target protein using the cofactor is expressed in the form fused with OmpC; (c) transforming the recombinant vector for expressing the target protein and the recombinant vector for regenerating the cofactor into E. coli; and (d) surface expressing the enzyme regenerating the cofactor and the target protein by culturing the transformed cell. 2. The method according to claim 1, wherein the recombinant vector for expression of the target protein and the recombinant vector for regeneration of the cofactor are compatible. 3. The method according to claim 1, wherein the cofactor is selected from the group consisting of dehydrogenase coenzyme, transferase coenzyme and isomerase and lyase coenzyme. 4. The method according to claim 3, wherein said dehydrogenase coenzyme is selected from the group consisting of nicotinamide-adenine-dinucleotide (NAD), nicotinamide-adenine-dinucleotide phosphate (NADP), flavin-mononucleotide (FMN), flavin-adenine-dinucleotide (FAD), cellular hemin and lipoic acid. 5. The method according to claim 3, wherein said transferase coenzyme is selected from the group consisting of adenosine triphosphate (ATP), phosphoadenyl sulfate (PAPS), uridine diphosphate (UDP), cytidine diphosphate (CDP), guanosine triphosphate (GTP), inosine triphosphate (ITP), coenzyme A (CoA), tetrahydrofol acid (CoF, FH.sub.4), biotin-thiamine pyrophosphate (TPP) and pyridoxal phosphate (PAL). 6. The method according to claim 3, wherein said isomerase and lyase coenzyme is selected from the group consisting of uridine diphosphate (UDP), pyridoxal phosphate (PAL), thiamine pyrophosphate (TPP) and vitamin B.sub.12. 7. The method according to claim 1, wherein the target protein is selected from the group consisting of hormones, hormone analogues, enzymes, enzyme inhibitors, signal transduction proteins or a part thereof, antibodies or a part thereof, single-chain antibodies, binding proteins, binding domains, peptides, antigens, adhesion proteins, structural proteins, regulatory proteins, toxic proteins, cytokine, transcription regulatory factors, blood coagulation factors and plant biodefense-inducing proteins. 8. The method according to claim 7, wherein the enzyme is selected from the group consisting of dehydrogenase, transferase, isomerase and lyase. 9. The method according to claim 8, wherein the cofactor is nicotinamide-adenine-dinucleotide (NAD) and the target protein is lactate dehydrogenase. 10. The method according to claim 9, wherein the cofactor regenerating enzyme is formate dehydrogenase. 11. The method according to claim 8, wherein the target protein is lactate dehydrogenase. 12. The method according to claim 11, wherein the cofactor regenerating enzyme is pyruvic acid dehydrogenase.

Details for Patent 7,897,366

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Recordati Rare Diseases, Inc.	PANHEMATIN	hemin for injection	For Injection	101246	07/20/1983		2026-04-27