Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing alpha or beta amino acid or substituted amino acid...
Patent
1989-12-22
1992-06-23
Griffin, Ronald W.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing alpha or beta amino acid or substituted amino acid...
43525232, 435840, 435843, C12P 1306, C12N 120, C12N 100
Patent
active
051242575
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
(1) Field of the Invention
The present invention relates to the microbiological industry, more particularly to methods of preparing aminoacids and, more specifically, to a fermentation method for preparing L-alanine
(2) Background of the Related Art
Known in the art are methods for preparing L-alanine by way of transformation from aspartic acid using immobilized cells of microorganisms possessing a .beta.-decarboxylase activity (U.S. Pat. No. 3,898,128).
Such known methods are multi-stage ones which is associated with the necessity of producing, for example, in the first stage of the process, L-aspartic acid from ammonium fumarate by transformation with the use of aspartase and, in the second stage of the process - transformation of the resulting L-aspartic acid into L-alanine by means of L-aspartate-.beta.-decarboxylase (Progress in Industrial Microbiology, v. 24, 1986, Tokyo, J. Chibata, T.Tosa, T. Kakimoto "Alanine", p. 224-232).
Known in the art is a method for recovering L-alanine for a racemic mixture, wherein D,L-alanine is subjected to acetylation and then, by means of an enzyme - aminoacylase, acetyl-L-alanine is selectively hydrolyzed to give L-alanine. Afterwards, L-alanine is separated from acetyl-D-alanine (U.S. Pat. No. 3,386,888).
This is a multi-stage process necessitating preparation of intermediate D,L-alanine, acetyl-D,L-alanine, aminoacylase enzyme, thus making the process more complicated and expensive.
SUMMARY OF THE INVENTION
The present invention, by using a novel type of mutants of microorganisms, provides a method for preparing L-alanine which would make it possible to simplify the process for obtaining the desired product in a commercially acceptable yield of the desired product in a culture liquid.
This problem is solved by a method for preparing L-alanine which utilizes culturing of microorganisms of the genus Brevibacterium and Corynebacterium on a nutrient medium containing assimilable sources of carbon, nitrogen, inorganic salts and organic compounds, stimulating growth of the microorganisms till accumulation of L-alanine in the cultural liquid and a subsequent recovery of the desired product. In accordance with the present invention, as the microorganisms use is made of mutants of microorganisms of the genus Brevibacterium and needing D-alanine for their growth.
Owing to the present invention it is now possible to prepare optically pure L-alanine in a culture liquid, avoiding previously indespensable stages associated with the recovery of L-alanine from a racemic mixture, the desired product content in the cultural liquid reaching 28 g/l.
According to the present invention, it is advisable to use the mutants of microorganisms of the species of Brevibacterium flavum deposited at the Vsesojuzny Nauchno-Issledovatel'sky Institut Genetiki i Selektsii Promyshlennykh Mikroorganizmov (VNIIGenetika), Moscow, Dorozhny proezd, 1 (All-Union Research Institute of Genetics and Selection of Industrial Microorganisms/VNIIGenetika/), the strain Brevibacterium flavum AAI registered under No. B-3061 on Apr. 5, 1984; the strain Brevibacterium flavum AA2 registered under No. B-3062 on Apr. 5, 1984.
For a further increase of the content of the desired product in the culture liquid (reaching 44/g/l), according to the present invention it is advisable that use be made of the mutants of microorganisms of the genus Brevibacterium resistant against D, L-.alpha.aminobutyric acid.
According to the present invention, it is desirable to use mutants of microorganisms of the species Brevibacterium flavum deposited at the Vsesojuzny Nauchno-Issledovatel'sky Institut Genetiki i Selektsil Promyshlennykh Mikroorganizmov (VNIIGenetika):
the strain Brevibacterium flavum AA5 registered under No. B-3991 on Apr. 1, 1987;
the strain Brevibacterium flavum AA6 registered under No. B-3992 on Apr. 1, 1987.
Furthermore, it is desirable, in accordance with the present invention, to use mutants of microorganisms of the species Corynebacterium glutaminicum deposited at the Vsesojuzny Nauchno
REFERENCES:
Zaitseva et al., Biological Abstracts, vol. 79(12), #103932.
Kase et al., Biological Abstracts, vol. 64(3), #14198.
Linde et al., Biological Abstracts, vol. 62(1), #1779.
Ambartsumian Artur A.
Ananikian Maritsa A.
Azizian Asmik G.
Kocharian Shavarsh M.
Griffin Ronald W.
Webber Pamela S.
LandOfFree
Method for preparing L-alanine does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Method for preparing L-alanine, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method for preparing L-alanine will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-932514