Gene encoding phosphoglucoisomerase

Details Gene encoding phosphoglucoisomerase Gene encoding phosphoglucoisomerase

2002-08-20

2004-01-20

Patterson, Jr., Charles L. (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Isomerase

Reexamination Certificate

active

06680190

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates, in general, to a method of producing L-amino acids and to a gene encoding phosphoglucoisomerase.
2. Background Information
Bacterial cells are used industrially to produce amino acids by fermentation processes (Ishino, S. et al.,
J. Gen. Appl. Microbiol
. 37:157-165 (1991), Kinoshita, S., Nakayama, K. and Nagasaki, S.,
J. Gen. Appl. Microbiol
. 4:128-129 (1958)). Although numerous research reports and reviews have appeared concerning fermentation processes and the mechanisms of accumulation of amino acids, more progress needs to be made to increase the yields of amino acids from microorganisms (Ishino, S. et al.,
J. Gen. Appl. Microbiol
. 37:157-165 (1991), Aida, K. et al., eds., “Biotechnology of Amino Acid Production,” Kodansha (Tokyo)/Elsevier (New York) (1986) and Marx, A. et al.,
Metabolic Engineering
1:35-48 (1999)).
There has been some success in using metabolic engineering to direct the flux of glucose derived carbons toward aromatic amino acid formation (Flores, N. et al.,
Nature Biotechnol
. 14:620-623 (1996)). However, the successful application in producer strains has not yet been documented (Berry, A.,
TIBTECH
14:250-256 (1996)).
Metabolic engineering relates to manipulation of the flow of carbons of starting materials, such as carbohydrates and organic acids, through the variety of metabolic pathways during fermentation. Studies have been done, for example, on the central metabolism of
Corynebacterium glutamicum
using
13
C NMR studies (Ishino, S. et al.,
J. Gen. Appl. Microbiol
. 37:157-165 (1991), Marx, A. et al.,
Biotechnology and Bioengineering
49:111-129 (1996)). Additionally, also using
13
C NMR, Walker et al. (Walker, T. et al,
J. Biol. Chem
. 257:1189-1195 (1982)) analyzed glutamic acid fermentation by
Microbacterium ammoniaphilum
, and Inbar et al (Inbar, L. et al.,
Eur. J. Biochem
. 149:601-607 (1985)) studied lysine fermentation by
Brevibacterium flavum.
The present invention solves a problem of improving yields of amino acids during fermentation using metabolic engineering.
SUMMARY OF THE INVENTION
The present invention provides a method of producing L-amino acids by culturing altered bacterial cells having increased amounts of NADPH as compared to unaltered bacterial cells, whereby L-amino acid yields from said altered bacterial cells are greater than yields from unaltered bacterial cells.
The present invention also provides a method of producing a bacterial cell with a mutated phosphoglucose isomerase (pgi) gene comprising (a) subcloning an internal region of the pgi gene into a suicide vector; and (b) inserting said suicide vector into a bacterial genome, via homologous recombination, whereby a bacterial cell with an altered pgi gene is produced. The invention further provides an altered bacterial cell produced according to this method.
The invention also provides a vector useful according to this method.
The present invention further provides isolated nucleic acid molecules comprising a polynucleotide encoding the
Corynebacterium glutamicum
phosphoglucose isomerase polypeptide having the amino acid sequence shown in
FIG. 1
(SEQ ID NO:2) or one of the amino acid sequence encoded by the DNA clone deposited in a bacterial host as NRRL Deposit Number B-30174 on Aug. 17, 1999.
The present invention also relates to recombinant vectors, which include the isolated nucleic acid molecules of the present invention, and to host cells containing the recombinant vectors, as well as to methods of making such vectors and host cells and for using them for production of Pgi polypeptides or peptides by recombinant techniques.
The invention further provides an isolated Pgi peptide having an amino acid sequence encoded by a polynucleotide described herein.
Further advantages of the present invention will be clear from the description that follows.


REFERENCES:
patent: 4086138 (1978-04-01), Kojuma et al.
patent: 5631150 (1997-05-01), Harkki et al.
patent: 0 733 712 (1996-09-01), None
patent: 0 780 477 (1997-06-01), None
patent: 1 087 015 (2001-03-01), None
patent: 2 772 788 (1999-06-01), None
patent: WO 94/10325 (1994-05-01), None
Berry, A., “Improving production of aromatic compounds inEscherichia coliby metabolic engineering,”TIBTECH 14:250-256, Elsevier Science Ltd. (1996).
Boles, E., et al., “The role of the NAD-dependent glutamate dehydrogenase in restoring growth on glucose of aSaccharomyces cerevisiaephophoglucose isomerase mutant,”Eur. J. Biochem. 271:469-477, Federation of European Biochemical Societies (1993).
Cocaign-Bousquet, M. and Lindley, N.D., “Pyruvate overflow and carbon flux within the central metabolic pathways ofCorynebacterium glutamicumduring growth on lactate,”Enzyme Microb. Tech. 17:260-267, Elsevier Science Inc. (1995).
Dialog File 351, Accession No. 12615741, Derwent WPI English language abstract for FR 2 772 788 (Document AO1) (1999).
Dominguez, H., et al., “Carbon-flux distribution in the central metabolic pathways ofCorynebacterium glutamicumduring growth on fructose,”Eur. J. Biochem. 254:96-102, Springer (May 1998).
Eikmanns, B.J., et al., “Cloning, Sequence Analysis, Expression, and Inactivation of theCorynebacterium glutamicum icdGene Encoding Isocitrate Dehydrogenase and Biochemical Characterization of the Enzyme,”J. Bacteriol. 177:774-782, American Society for Microbiology (1995).
Fitzpatrick, R., et al., “Construction and characterization of recA mutant strains ofCorynebacterium glutamicumandBrevibacterium lactofermentum,”Appl. Microbiol. Biotechnol. 42:575-580, Springer-Verlag (1994).
Flores, N., et al., “Pathway engineering for the production of aromatic compounds inEscherichia coli,”Nature Biotechnol. 14:620-623, Nature Publishing Co. (1996).
González Siso, M.I. et al., “Reoxidation of the NADPH produced by the pentose phosphate pathway is necessary for the utilization of glucose byKluyveromyces lactis rag2mutants,”FEBS Lett. 387:7-10, Federation of European Biochemical Societies (1996).
Inbar, L., et al., “Natural-abundance13C nuclear magnetic resonance studies of regulation and overproduction of L-lysine byBrevibacterium flavum,”Eur. J. Biochem. 149:601-607, Springer International (1985).
International Search Report for International Application No. PCT/US00/19914, mailed Dec. 28, 2000.
Ishino, S., et al., “13C Nuclear Magnetic Resonance Studies of Glucose Metabolism in L-Glutamic Acid and L-Lysine Fermentation byCorynebacterium Glutamicum,”J. Gen. Appl. Microbiol. 37:157-165, The Microbiology Research Foundation (1991).
Kinoshita, S., et al., “L-Lysine Production Using Microbial Auxotroph ,”J. Gen. Appl. Microbiol. 4:128-129, The Institute of Applied Microbiology (1958).
Marx, A., et al., “Determination of the Fluxes in the Central Metabolism ofCorynebacterium glutamicumby Nuclear Magentic Resonance Spectroscropy Combined with Metabolite Balancing,”Biotechnol. Bioengineering 49:111-129, Wiley (1996).
Marx, A., et al., “Response of the Central Metabolism inCorynebacterium glutamicumto the use of an NADH-Dependent Glutamate Dehydrogenase,”Metabolic Engineering 1:35-48, Academic Press (Jan. 1999).
Moritz, B., et al., “Kinetic properties of the glucose-6-phosphate and 6-phosphogluconate dehyrogenases fromCorynebacterium glutamicumand their application for predicting pentose phosphate pathway flux in vivo,”Eur. J. Biochem. 267:3442-3452, Federation of European Biochemical Societies (Jun. 2000).
O'Gara, J.P. and Dunican, L.K., “Direct Evidence for a Constitutive Internal Promoter in the Tryptophan Operon ofCorynebacterium glutamicum,”Biochem. &Biophys. Res. Comm. 203:820-827, Academic Press (1994).
Patent Abstracts of Japan, Publication No. 09224661, published Sep. 2, 1997.
Patent Abstracts of Japan, Publication No. 09224662, published Sep. 2, 1997.
Sahm, H., et al., “Construction of L-Lysine-, L-Threonine-, or L-Isoleucine-Overproducing Strains ofCorynebacterium glutamicum,”Ann. N. Y. Acad. Sci. 782:25-39, Springer Press (1996).
Shi, H., et al., “Effect of Modifying Metabolic Network on Poly-3-Hydroxybutyrate Biosynthes

Affiliated with

Also associated with

Rating

Gene encoding phosphoglucoisomerase does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Gene encoding phosphoglucoisomerase, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Gene encoding phosphoglucoisomerase will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3194103