PROCESS AND MATERIALS FOR PRODUCTION OF GLUCOSAMINE AND N-ACETYLGLUCOSAMINE
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Abstract
A biosynthetic method for producing glucosamine and N-acetylglucosamine is disclosed. Such a method includes the fermentation of a genetically modified microorganism to produce glucosamine and/or N-acetylglucosamine. Also disclosed are genetically modified microorganisms that are useful for producing glucosamine and N-acetylglucosamine. In addition, methods of recovering N-acetylglucosamine that has been produced by a fermentation process, including methods that result in N-acetylglucosamine of high purity, are described. Also disclosed is a method to produce glucosamine from N-acetylglucosamine.
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Citations
279 Claims
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1-212. -212. (canceled)
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213. A method to produce N-acetylglucosamine-6-phosphate or N-acetylglucosamine by fermentation, comprising:
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a) culturing in a fermentation medium a microorganism that is transformed with at least one recombinant nucleic acid molecule comprising a nucleic acid sequence encoding a glucosamine-6-phosphate acetyltransferase that has an amino acid sequence that is at least 95% identical to SEQ ID NO;
32; andb) collecting a product produced from the step of culturing which is selected from the group consisting of N-acetylglucosamine-6-phosphate and N-acetylglucosamine. - View Dependent Claims (214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249)
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250. A method to produce N-acetylglucosamine-6-phosphate, N-acetylglucosamine, glucosamine or glucosamine HCl by fermentation, comprising:
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culturing in a fermentation medium a microorganism that is transformed with at least one recombinant nucleic acid molecule comprising a nucleic acid sequence encoding a glucosamine-6-phosphate acetyltransferase that has an amino acid sequence that is at least 95% identical to SEQ ID NO;
32; andcollecting a product produced from the step of culturing which is selected from the group consisting of N-acetylglucosamine-6-phosphate and N-acetylglucosamine; and wherein the method further comprises a step selected from the group consisting of; a) purifying N-acetylglucosamine from the fermentation medium; b) dephosphorylating N-acetylglucosamine-6-phosphate to produce N-acetylglucosamine; and c) treating N-acetylglucosamine to produce a glucosamine product selected from the group consisting of;
glucosamine and glucosamine HCl. - View Dependent Claims (251)
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252. A method to produce N-acetylglucosamine-6-phosphate or N-acetylglucosamine by fermentation, comprising:
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a) culturing in a fermentation medium a microorganism that expresses; i) a recombinant nucleic acid molecule encoding a glucosamine-6-phosphate acetyltransferase that has an amino acid sequence that is at least 95% identical to SEQ ID NO;
32; andii) a recombinant nucleic acid molecule encoding a glucosamine-6-phosphate synthase that has an amino acid sequence that is at least 95% identical to the amino acid sequence selected from the group consisting of;
SEQ ID NO;
4, SEQ ID NO;
6, SEQ ID NO;
8, SEQ ID NO;
10, SEQ ID NO;
12, SEQ ID NO;
14, SEQ ID NO;
16, SEQ ID NO;
18, and SEQ ID NO;
20; andb) collecting a product produced from the step of culturing which is selected from the group consisting of N-acetylglucosamine-6-phosphate and N-acetylglucosamine. - View Dependent Claims (253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271)
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272. A method to produce N-acetylglucosamine-6-phosphate or N-acetylglucosamine by fermentation, comprising:
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a) culturing in a fermentation medium an E. coli that expresses; i) a recombinant nucleic acid molecule encoding a glucosamine-6-phosphate acetyltransferase that is at least 95% identical to the amino acid sequence of SEQ ID NO;
32; andii) a recombinant nucleic acid molecule encoding a glucosamine-6-phosphate synthase that is at least 95% identical to the amino acid sequence of SEQ ID NO;
6; andb) collecting a product produced from the step of culturing which is selected from the group consisting of N-acetylglucosamine-6-phosphate and N-acetylglucosamine. - View Dependent Claims (273, 274, 275, 276, 277, 278, 279)
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Specification