Isolation of the biosynthesis genes for pseudo-oligosaccharides from streptomyces glaucescens GLA.O, and their use
First Claim
1. An isolated polynucleotide comprising a nucleotide sequence which encodes a protein for biosynthesizing acarbose, wherein said nucleotide sequence is capable of hybridizing under stringent conditions with the sequence of SEQ ID No:
- 7.
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Accused Products
Abstract
The invention relates to a recombinant DNA molecule which comprises genes for biosynthesizing acarbose and homologous pseudo-oligosaccharides; to oligonucleotide primers for the PCR amplification of the molecule; to proteins which can be obtained by expressing the genes located on a molecule; to vectors and host cells which comprise the above-mentioned DNA molecule; to proteins which are encoded by the DNA molecule; to proteins which are expressed by means of said vectors in said host cells; to processes for preparing acarbose by introducing the characterized genes into appropriate host organisms and/or eliminating these genes from the host organisms; to processes for completing the gene cluster of genes for biosynthesizing acarbose, to processes for isolating analogous gene clusters in organisms other than Streptomyces glaucescens GLA.O, to processes for mutating promoters of endogenous acarbose biosynthesis genes for the purpose of increasing the yield of acarbose, to the use of Streptomyces glaucescens GLA.O for preparing acarbose and for preparing mutants of Streptomyces glaucescens GLA.O which are optimized with regard to the acarbose yield.
18 Citations
26 Claims
- 1. An isolated polynucleotide comprising a nucleotide sequence which encodes a protein for biosynthesizing acarbose, wherein said nucleotide sequence is capable of hybridizing under stringent conditions with the sequence of SEQ ID No:
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5. An isolated polynucleotide comprising a nucleotide sequence encoding at least one protein for biosynthesizing acarbose, wherein said nucleotide sequence is selected from the group consisting of:
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(a) a nucleotide sequence of SEQ ID No;
7;
(b) a nucleotide sequence which is capable of hybridizing, under stringent conditions, with the sequence of SEQ ID No;
7;
(c) a nucleotide sequence consisting of nucleotide 1 to 720 of SEQ ID No;
7;
(d) a nucleotide sequence consisting of nucleotides 720 to 2006 of SEQ ID No;
7;
(e) a nucleotide sequence consisting of nucleotides 2268 to 3332 of SEQ ID No;
7;
(f) a nucleotide sequence consisting of nucleotides 3332 to 4306 of SEQ ID No;
7;
(g) a nucleotide sequence consisting of nucleotides 4380 to 5414 of SEQ ID No;
7;
(h) a nucleotide sequence consisting of nucleotides 5676 to 6854 of SEQ ID No;
7;
(i) a nucleotide sequence which is capable of hybridizing, under stringent conditions, with the sequence according to (a), (b), (c), (d) (e), (f), (g) or (h); and
(j) a nucleotide sequence which, because of the degeneracy of the genetic code, differs from the nucleotide sequence according to (a), (b), (c), (d), (e), (f), (g) (h) and (i), but which permits the expression of any protein which can be expressed using the nucleotide sequence of (a), (b), (c), (d), (e), (f), (g), (h) and (i), respectively. - View Dependent Claims (6, 7, 8, 9, 10, 11, 12, 13)
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25. An isolated polynucleotide comprising a nucleotide sequence encoding a protein for biosynthesizing acarbose, wherein said nucleotide sequence is capable of hybridizing, under stringent conditions, with a nucleotide sequence selected from the group consisting of a nucleotide sequence consisting of nucleotides 1 to 720 of SEQ ID NO:
- 7, a nucleotide sequence consisting of nucleotides 720 to 2006 of SEQ ID NO;
7, a nucleotide sequence consisting of nucleotides 2268 to 3332 of SEQ ID NO;
7, a nucleotide sequence consisting of nucleotides 3332 to 4306 of SEQ ID NO;
7, a nucleotide sequence consisting of nucleotides 4380 to 5414 of SEQ ID NO;
7, and a nucleotide sequence consisting of nucleotides 5676 to 6854 of SEQ ID NO;
7.
- 7, a nucleotide sequence consisting of nucleotides 720 to 2006 of SEQ ID NO;
Specification