Process and materials for production of glucosamine
AssigneeArkion Life Sciences, LLC
InventorsAlan Berry, Richard P. Burlingame, James R. Millis
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Abstract
The present invention relates to a method and materials for producing glucosamine by fermentation of a genetically modified microorganism. Included in the present invention are genetically modified microorganisms useful in the present method for producing glucsamine, as well as recombinant nucleic acid molecules and the proteins produces by such recombinant nucleic acid molecules.
Claims59
A method to produce glucosamine by fermentation, comprising:
- a. culturing in a fermentation medium comprising assimilable sources of carbon, nitrogen and phosphate, a microorganism which comprises at least one genetic modification that increases the activity of glucosamine-6-phosphate synthase, wherein said genetic modification is selected from the group consisting of:
- i. transformation of said microorganisn with a recombinant nucleic acid molecule encoding glucosamine-6-phosphate synthase from E. coli which has glucosamine-6-phosphate synthase activity;
- and
- ii. genetic modification of a gene encoding glucosamine-6-phosphate synthase from E. coli that increases the activity of said glucosamine-6-phosphate synthase, wherein said genetic modification results in at least one nucleic acid modification selected from the group consisting of deletion, insertion, and substitution of at least one nucleotide of said gene encoding glucosamine-6-phosphate synthase, said at least one nucleotide modification resulting in increased glucosamine-6-phosphate synthase activity;
- wherein said step of culturing produces and accumulates a product selected from the group consistng of glucosamine-6-phosphate and glucosamine from said microorganism;
- and
- b. recovering and purifying said product.
A method to produce glucosamine by fermentation, comprising:
- a. culturing in a fermentation medium comprising assimilable sources of carbon, nitrogen and phosphate, an Escherichia coli transformed with a recombinant nucleic acid molecule encoding glucosamine-6-phosphate synthase from E. coli, wherein said recombinant nucleic acid molecule increases glucosamine-6-phosphate synthase activity in said Escherichia coli, and wherein said recombinant nucleic acid molecule is operatively linked to a transcription control sequence;
- wherein said step of culturing produces and accumulates a product selected from the group consisting of glucosamine-6-phosphate and glucosamine from said Escherichia coli;
- and
- b. recovering and purifing said product.
A microorganism for producing glucosamine by a biosynthetic process, said microorganism being transformed with a recombinant nucleic acid molecule comprising a nucleic acid sequence encoding glucosamine-6-phosphate synthase from E. coli, said nucleic acid sequence being operatively linked to a transcription control sequence and comprising a genetic modification which increases the activity of said glucosamine-6-phosphate synthase;
- wherein expression of said nucleic acid sequence increases production of glucosamine by said microorganism.
A microorganism for producing glucosamine by a biosynthetic process, said microorganism comprising:
- (a) a recombinant nucleic acid molecule encoding glucosamine-6-phosphate synthase from E. coli, said recombinant nucleic acid molecule being operatively linked to a transcription control sequence, wherein expression of said recombinant nucleic acid molecule increases the activity of glucosamine-6-phosphate synthase by said microorganism;
- and,
- (b) at least one genetic modification in a gene encoding a protein selected from the group consisting of N-acetylglucosamine-6-phosphate deacetylase, glucosamine-6-phosphate deaminase, N-acetyl-glucosamine-specific enzyme IINag and EIIM,P/IIIMan of the PEP:mannose PTS, wherein said genetic modification decrease the activity of said protein.
A recombinant nucleic acid molecule comprising a nucleic acid sequence encoding a glucosamine-6-phosphate synthase from E. coli having a genetic modification that results in increased glucosamine-6-phosphate synthase activity.
A method to produce glucosamine by fermentation, comprising:
- a. culturing in a fermentation medium comprising assimilable sources of carbon, nitrogen and phosphate, a microorganism having at least one genetic modification that increases the activity of glucosamine-6-phosphate synthase wherein said genetic modification is selected from the group consisting of:
- i. transformation of said microorganism with a recombinant nucleic acid molecule encoding glucosamine-6-phosphate synthase from E. coli which has glucosamine-6-phosphate synthase activity;
- and
- ii. genetic modification of a gene encoding glucosamine-6-phosphate synthase from E. coli that increases the activity of said glucosamine-6-phosphate synthase, and wherein said genetically modified microorganism is produced by a process comprising the steps of:
- (1) generating modifications in an isolated nucleic acid molecule comprising a nucleic acid sequence encoding glucosamine-6-phosphate synthase from E. coli to create a plurality of modified nucleic acid sequences;
- (2) transforming microorganisms with said modified nucleic acid sequences to produce genetically modified microorganisms;
- (3) screening said genetically modified microorganisms for glucosamine-6-phosphate synthase activity;
- and,
- (4) selecting said genetically modified microorganisms which have increased glucosamine-6-phosphate synthase activity;
- wherein said step of culturing produces a product selected from the group consisting of glucosamine-6-phosphate and glucosamine from said microorganism;
- and,
- b. recovering said product.
The method of clam 7, wherein said step of culturing is performed so that an excess of glucose is maintained.
Dependencies are inferred from the claim text.