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EP0077196A2 Expired / Lapsed EP ⏱️ 0.0 years remaining

Aromatic amino acid-producing microorganisms

Assignee: Genex Corp
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Filing: 1982-10-08
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Granted: —
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Expires: 2002-10-08
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Abstract

Abstract An isolated microbial gene that specifies the biosynthesis of a 3-deoxy-2-keto-D-arabinoheptulosonic acid-7-phosphate (DAHP) synthetase, which is resistant to feedback inhibition by aromatic amino acids; plasmids containing such a gene; and microorganisms transformed thereby.

Inventors

  • 👤 Richard A. Synenki

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-09 09:27
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/93; C12N15/00; C12N15/09; C12N15/11; C12N15/52; C12P; C12P13/00; C12P13/04; C12P13/22

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 26 words
A substantially isolated intact microbial gene that specifies the biosynthesis of a 3-deoxy-2-keto-D-arabinoheptulosonic acid-7-phosphate (DAHP) synthetase, which is resistant to feedback inhibition by aromatic amino acids.
Claim 2
Dependent Claim 26 words
A substantially isolated intact microbial gene that specifies the biosynthesis of a 3-deoxy-2-keto-D-arabinoheptulosonic acid-7-phosphate (DAHP) synthetase, which is resistant to feedback inhibition by aromatic amino acids.
Claim 3
Dependent Claim 12 words
The microbial gene of claim 1, comprising the following deoxyribonucleotide sequence: .
Claim 4
Dependent Claim 192 words
wherein, the 5' to 3' strand, beginning with the amine terminus and the amino acids for which each triplet codes are shown, and wherein within each triplet, A is deoxyadenyl T is thymidyl G is deoxyguanyl C is' deoxycytosyl . X is A, T, C or G Y is T or C When Y is C, Z is A or G H is A, T or C Q is T or A When Q is T, R is C and S is A, T, C or G When Q is A, R is G and S is T or C M is A or G L is A or C When L is A, N is A or G When L is C, N is A, T, C or G. Gly is glycine Ala is alanine Val is valine Leu is leucine Ile is isoleucine Ser is serine Thr is threonine Phe is phenylalanine Tyr is tyrosine Trp is tryptophan Cys is cysteine Met is methionine Asp is aspartic acid Glu is gluatmic acid Lys is lysine ARg is arginine His is histidine Pro is proline Gln is glutamine Asn is asparagine
Claim 5
Dependent Claim 192 words
wherein, the 5' to 3' strand, beginning with the amine terminus and the amino acids for which each triplet codes are shown, and wherein within each triplet, A is deoxyadenyl T is thymidyl G is deoxyguanyl C is' deoxycytosyl . X is A, T, C or G Y is T or C When Y is C, Z is A or G H is A, T or C Q is T or A When Q is T, R is C and S is A, T, C or G When Q is A, R is G and S is T or C M is A or G L is A or C When L is A, N is A or G When L is C, N is A, T, C or G. Gly is glycine Ala is alanine Val is valine Leu is leucine Ile is isoleucine Ser is serine Thr is threonine Phe is phenylalanine Tyr is tyrosine Trp is tryptophan Cys is cysteine Met is methionine Asp is aspartic acid Glu is gluatmic acid Lys is lysine ARg is arginine His is histidine Pro is proline Gln is glutamine Asn is asparagine
Claim 6
Dependent Claim 11 words
The microbial gene of claim 1, comprising the following deoxyribonucleotide sequence:
Claim 7
Dependent Claim 20 words
The gene of claim 1, wherein the gene is a feedback resistant aroG, aroF or aroH gene of bacterial origin.
Claim 8
Dependent Claim 19 words
A plasmid capable of replication in a bacterial cell, comprising the gene of claims 1, 2, 3 or 4.
Claim 9
Dependent Claim 12 words
The plasmid of claim S, comprising the bacterial plasmid pMF-3 or pBR322.
Claim 10
Dependent Claim 18 words
A microorganism transformed with the plasmid of claim 5, having the ability to maintain and replicate said plasmid.
Claim 11
Dependent Claim 50 words
The microorganism of claim 7, wherein said plasmid includes signals recognizable by said microorganism which control the expression of the feedback resistant - DAHP synthetase gene, whereby said microorganism has the ability to produce DAHP synthetase which is active in the presence of normally inhibiting amounts of aromatic amino acids.
Claim 12
Independent Claim 77 words
A method for cloning a DNA segment which comprises first, inserting said segment into a single- copy plasmid to produce a first recombinant plasmid, transforming microbial cells with said first recombinant plasmid, isolating plasmid DNA from said microbial cells and excising said DNA segment or a portion thereof from said plasmid DNA and, second, inserting said excised segment into a multi-copy plasmid to form a second recombinant plasmid and transforming microbial cells with said second recombinant plasmid.
Claim 13
Independent Claim 77 words
A method for cloning a DNA segment which comprises first, inserting said segment into a single- copy plasmid to produce a first recombinant plasmid, transforming microbial cells with said first recombinant plasmid, isolating plasmid DNA from said microbial cells and excising said DNA segment or a portion thereof from said plasmid DNA and, second, inserting said excised segment into a multi-copy plasmid to form a second recombinant plasmid and transforming microbial cells with said second recombinant plasmid.
Claim 14
Dependent Claim 19 words
The method of claim 9 wherein the DNA segment to be cloned contains a feedback resistant DAHP synthetase gene.
Claim 15
Dependent Claim 22 words
A method of producing an aromatic amino acid comprising culturing a microorganism as claimed in claim 8 and recovering said amino acid.

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