Process for producing 4-hydroxy-L-proline

Details Process for producing 4-hydroxy-L-proline Process for producing 4-hydroxy-L-proline

1993-04-26

1994-12-20

Wax, Robert A.

Chemistry: molecular biology and microbiology

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

Preparing alpha or beta amino acid or substituted amino acid...

4351723, 43525233, 4353201, C12P 1324, C12N 1509, C12N 1570

Patent

active

053745426

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

The present invention relates to a process for producing 4-hydroxy-L-proline by use of a microorganism capable of biosynthesizing trans-4-hydroxy-L-proline and/or cis-4-hydroxy-L-proline (hereinafter referred to as 4-hydroxy-L-proline) from D- or L-4-hydroxy-2-oxoglutaric acid (hereinafter referred to as 4-hydroxy-2-oxoglutaric acid). 4-Hydroxy-L-proline is an amino acid which is useful as an intermediate for the synthesis of pharmaceuticals, etc.
1. Background of the Invention
As the hitherto known methods for the production of 4-hydroxy-L-proline, mention may be made of a method comprising hydrolyzing collagen and recovering, from the hydrolyzed collagen-constituting amino acids, trans-4-hydroxy-L-proline; a method for synthesizing trans-4-hydroxy-L-proline from D-glutamic acid (Bulletin of the Chemical Society of Japan, 47., 1704, 1974); and a method for synthesizing trans-4-hydroxy-L-proline from glyoxal and oxaloacetic acid (Journal of Organic Chemistry, 42, 3440, 1977) are known.
The prior art methods are not satisfactory for the industrial production because of (1) high cost of raw materials, (2) many reaction steps involved and (3) problems in the recovering and purifying steps. Thus, an improved method is desired for the industrially inexpensive production of 4-hydroxy-L-proline.
2. Disclosure of the Invention
According to the present invention, 4-hydroxy-L-proline is efficiently produced by converting 4-hydroxy-2-oxoglutaric acid into 4-hydroxy-L-proline in an aqueous solution in the presence of an amino group donor and an enzyme source capable of converting 4-hydroxy-2-oxoglutaric acid into 4-hydroxy-L-proline.
Specifically, the conversion reaction of 4-hydroxy2-oxoglutaric acid into 4-hydroxy-L-proline is carried out by bringing the above-mentioned enzyme source into contact with 4-hydroxy-2-oxoglutaric acid and an amino group donor in an aqueous solution having a pH suitable for the enzyme reaction, such as a buffer and a physiological saline.
As the enzyme source to be used in the present invention, either of a purified enzyme preparation or a crude enzyme preparation may be used. Preferably, a microorganism having an enzymatic activity of converting 4-hydroxy-2oxoglutaric acid into 4-hydroxy-L-proline may be used. More specifically, the microorganism having such an activity may be used in a form of a culture, cells or processed cells.
When a microorganism is used as the enzyme source, 4-hydroxy-2-oxoglutaric acid is brought into contact with the culture obtained by culturing the microorganism, during the culturing, in order to form 4-hydroxy-L-proline in the culture.
Any microorganism can be used in the present invention so long as it has an enzymatic activity to convert 4-hydroxy-2-glutaric acid into 4-hydroxy-L-proline. As the microorganism having such enzyme activity, mention may be made of those belonging to the genus Escherichia. As the practical strain, Escherichia coli ATCC33625 is mentioned.
Mutants and genetically engineered transformants in which enzyme activity for L-proline biosynthesis has been intensified, by a recombinant DNA technology and conventional mutagenesis, etc. are preferably used. For example, a microorganism possessing .gamma.-glutamylkinase activity of which feedback inhibition by L-proline has been released, is mentioned.
The genetically engineered transformants with the intensified enzyme activity include, for example, Escherichia coli K83. The strain carries a recombinant DNA comprising a proB gene derived from Escherichia coli MM294 strain which gene codes for .gamma.-glutamylkinase, of which feedback inhibition caused by L-proline has been released. Such recombinant DNA-carrying strains may be produced by the method of Deutche, et al. (Nucleic Acid Research, 12, 6337, 1984) or by the methods described in the Examples in the present specification.
Escherichia coli K83 has been deposited with the Fermentation Research Agency, Industrial Science and Technology in Japan as of March 15, 1990, in terms of the Budapest Treaty, under FERM BP-2

REFERENCES:
patent: 3905867 (1975-09-01), Kurimura et al.
patent: 4421853 (1983-12-01), Updike et al.
patent: 4463094 (1984-07-01), Chibata et al.
patent: 4594323 (1986-06-01), Csonka et al.
patent: 4681852 (1987-07-01), Tribe
patent: 4707449 (1987-11-01), Shay
patent: 5017482 (1992-05-01), Katsumata et al.
patent: 5087566 (1992-02-01), Takano et al.
Garcia, J. L., et al., 1985, Microbiologia, 1(1):43-51.
Csonka, L. N., et al., 1988, Gene, 64(2):199-205.
Dandekar, A. M., et al., 1988, Journal of Bacteriology, 170(12):5943-5945.
Jakowec, M. W., et al., 1985, Applied and Environmental Microbiology, 50(2):441-446.
Smith, L. T., 1985, Journal of Bacteriology, 164(3):1088-1093.
Smith, C. J., et al., 1984, Journal of Bacteriology, 157(2):545-551.
Eguchi, L., et al, 1974, Bulletin of the Chemical Society of Japan 47(7):1704-1708.
Ramaswamy, S. G., et al., 1977, Journal of Organic Chemistry 42(21):3440-3442.

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