Riboflavin-biosynthesis in fungi

Details Riboflavin-biosynthesis in fungi Riboflavin-biosynthesis in fungi

1996-09-24

1998-10-13

Wax, Robert A.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 66, 4352541, 43525411, 4352542, 43525421, 43525422, 4353201, 435325, 536 232, 935 22, C07H 2104, C12N 1500, C12N 114, C12P 1940

Patent

active

058210903

DESCRIPTION:

BRIEF SUMMARY
This application is national stage application of PCT/EP95/00958, filed on Mar. 15, 1995.
The present invention relates to the genes for riboflavin biosynthesis in fungi, to the proteins encoded thereby and to genetic engineering processes for preparing riboflavin using these genes and gene products.
The preparation of riboflavin by fermentation of fungi such as Eremothecium ashbyii or Ashbya gossypii has been disclosed (The Merck Index, Windholz et al., eds. Merck & Co., page 1183 (1983)).
EP 405370 describes riboflavin-overproducing bacterial strains which have been obtained by transformation of the riboflavin bio- synthesis genes from Bacillus subtilis.
Since the genetics of riboflavin biosynthesis in bacteria and eu- karyotes differ, the abovementioned genes from Bacillus subtilis are unsuitable for a recombinant process for preparing riboflavin using eukaryotic producer organisms such as Ashbya gossypii.
The cloning of the riboflavin biosynthesis genes of the yeast Saccharomyces cerevisiae was described in a patent application filed at the German Patent Office on Nov. 19, 1992.
However, it was not possible to clone the Ashbya gossypii ribo- flavin biosynthesis genes using the S. cerevisiae rib genes by conventional hybridization methods; evidently the homology of the rib genes from S. cerevisiae and A. gossypii was not great enough for hybridization.
It is an object of the present invention to isolate the ribo- flavin biosynthesis genes from a eukaryote in order in this way to provide a recombinant process for preparing riboflavin in a eukaryotic producer organism.
We have found that this object is achieved by isolation of six genes (rib genes) found in the ascomycete Ashbya gossypii which code for enzymes of riboflavin biosynthesis starting from GTP.
The invention relates to the following DNA sequences:
DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 2, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 2, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.
DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 4, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 4, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.
DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 6, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 6, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.
DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 8, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 8, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.
DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 10, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 10, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.
DNA sequences which code for a polypeptide with the amino-acid sequence depicted in SEQ ID NO: 12, or for an analog or derivative of the polypeptide shown in SEQ ID NO: 12, in which one or more amino acids have been deleted, added or replaced by other amino acids, without essentially reducing the enzymatic action of the polypeptide.
The genes and their gene products (polypeptides) are shown in the sequence listing with their primary structure and are assigned as follows:
Guanosine triphosphate (GTP) is converted by GTP cyclohydrolase II (rib 1 gene product) into 2,5-

REFERENCES:
Ngo et al. (1994) Computational Complexity, Protein Stucture Prediction, and the Levinthal Paradox. In: The Protein Folding Problem and Tertiary Structure Prediction. Eds. Merz et al., Birkhauser, Boston, MA, pp. 491-495, Jan. 1994.
Buitrago et al., Yeast, vol. 9, No. 10, Oct. 1993, pp. 1099-1102.
Diognon et al., Yeast, vol. 9, No. 2, 1993, pp. 189-199.
The Merck Index, pp. 1183 and 1983.
Watson et al., Molcular Biology of the Gene, 4.sup.th Ed., vol. 1, 1987, pp. 86-87 and 437-440.
Moore et al., Nature Biotech., vol. 14, 1996, pp. 458-467).
SPEE et al., Nucleic Acids Research, 1993, vol. 21, No. 3, 1993, pp. 777-778.
Rellos et al., Prot. Expres. and Purif., vol. 5, 1994, pp. 270-277.
Mehta et al., Indian Journal of Experimental Biology, vol. 18, Mar. 1980, pp. 243-244.
Hollander et al., Biochem. and Biophys. Res. Comm., vol. 89, 1979, pp. 759-763.
Richter et al., J. og Bacteriology, vol. 174, 1992, pp. 4050-4056.
Winestock et al., J. of Biol. Chem., vol. 238, No. 8, Aug. 1963, pp. 2866-2874.

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