Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – Nonplant protein is expressed from the polynucleotide
Patent
1996-05-09
1999-08-03
Smith, Lynette F.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
Nonplant protein is expressed from the polynucleotide
800298, 800278, 800288, 4353201, 435419, 435468, 536236, 536 237, 536 241, C12N 1500, C12N 1582, C12N 1529, A01H 400
Patent
active
059327812
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a DNA coding for an enzyme involved in the biosynthesis of ectoine, a method for giving ectoine synthetic ability to a host cell by introducing said DNA thereto, and a transformed host cell.
PRIOR ART
Under an environment of a high osmotic pressure, a certain microorganisms can obtain a tolerance to the surrounding stress by accumulating so-called "compatible solute" within their cells. It is known that the compatible solute includes saccharides, polyols, betaines or a certain amino acids (cf. Truper, H. G. et al., Experientia, Vol. 42, pp. 1182-1187, 1986).
Ectoine is a cyclic amino acid, which is 1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid or 3,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid. Said ectoine has been found as a compatible solute which is produced by a halophilic microorganism, Ectothiorhodospira halochloris, and it is further known that it has a tolerance to a high osmotic pressure ("high osmotic 135-139, 1985; Mitsuo Takano et al., a program for Symposium of The Japan Fermentation Engineering Association, p. 193, 1988!. There are also some reports as to the biosynthetic pathway of ectoine (cf. Peters, P. et al., FEMS Microbiol. Lett., Vol. 71, pp. 157-162, 1990) and as to the physical properties thereof (cf. Khunajakr, N., et al., Annual Reports of International Center of Cooperative Research in Biotechnology, Japan, Vol. 12, pp. 157-167, 1989).
Ectoine is biosynthetized from L-aspartate-.beta.-semialdehyde, in which steps L-diaminobutyric acid trans-aminase, L-diaminobutyric acetyltransferase, and ectoine synthase are involved. Hereinafter, these enzymes are generally called as "ectoine synthetases". As a system being capable of inducing the biosynthesis of an enzyme by bacteria, there is known so-called "operon" which is a regulatory system for expression of a gene, where a series of enzymes are synchronously induced by a single regulatory gene. The ectoine synthase is an enzyme which can synthetize ectoine from a-N-acetyl-diaminobutyric acid, and there are reports concerning a method for isolating and purifying said enzyme from a halophilic microorganism and the properties thereof (cf. Mihoko Yamamoto, et al., Summary of Symposium of Japan Biotechnology Society, p. 203, 1992). However, the gene structure and nucleotide sequence of the ectoine synthase and the ectoine synthetases have never been known.
On the basis of a knowledge that a certain micro-organism can accumulate ectoine within the cells thereof and thereby can grow even under an environment condition of strong stress to living bodies (e.g. high osmotic pressure), it has been proposed a method for extracting and isolating ectoine by aiming at the function of ectoine (cf. Khunajakr, N. et al., Annual Reports of International Center of Cooperative Research in Biotechnology, Japan, Vol. 12, pp. 157-167, 1989) and a method for chemical synthesis of ectoine (cf. JP-A-3-31265).
DISCLOSURE OF THE INVENTION
Aiming at the specific function of the ectoine, the present inventors have intensively investigated the ectoine from the following viewpoints. Where a high osmotic tolerance is given to, for example, a microorganism or a plant by giving an ability of biosynthesis of ectoine, not to ectoine per se, it will be able to develop a method for the efficient production of an useful product by a fermentation in a high concentration, and further it will be able to create a plant having a resistance to drought and having tolerance to a high osmotic pressure due to the droughty circumstance. Moreover, where the microorganisms are grown in a droughty ground such as in a desert, it will be also possible to change to a fertilized soil and further to create a plant which is suitable for planting in a droughty ground and at highly salty environment such as at a seaside.
BRIEF DESCRIPTION OF DRAWING
FIG. 1 shows a restriction endonuclease map of a DNA of a microorganism of the genus Halomonas, KS-3 strain, The signals in said figure mean the sites to be cleaved by each restriction
REFERENCES:
Yamamoto et al., "Summary of Symposium of Japan Biotechnology Society," p. 203 (1992).
Ono Hisayo
Takano Mitsuo
Yamada Hiroyuki
Yamatoya Kazuhiko
Dainippon Pharmaceutical Co., Ltd.
Haas Thomas
Smith Lynette F.
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