Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1997-02-07
1998-06-09
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
43525411, 4351721, 435203, 530317, C12P 2102, C12N 930, C12N 1509, C07K 512
Patent
active
057632219
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a transformant producing substance PF1022, which is a cyclic depsipeptide, a process for producing the substance PF1022, a method for elevating the productivity of the substance PF1022, and a method for transforming a microorganism belonging to the class Hyphomycetes.
2. Description of the Related Art
Microorganisms belonging to the class Hyphomycetes produce as metabolites various useful substances such as antibiotics, physiologically active substances and enzymes. Accordingly, for a long time, there have been studied and developed methods for obtaining such products by culturing these microorganisms on a large scale. As an example of common techniques for efficiently obtaining substances produced by microorganisms, a method which comprises artificially constructing mutants by, for example, UV irradiation or the use of a mutagenic agent and selecting a strain capable of producing the target substance in a large amount from among the mutants thus obtained is cited.
A strain newly constructed by such a method (hereinafter sometimes referred to as a highly productive strain) cannot always produce the desired metabolite at a high productivity in the same medium and under the same culture conditions as those for the parent strain thereof. Even though a highly productive strain is obtained by the above-mentioned technique, therefore, it is necessary to study, for every strain, medium and culture conditions suitable for the strain. In the case where a large fermentation tank is employed, in particular, the yield of the fermentation product widely varies depending on the medium and culture conditions, even when a highly productive strain which has been bred by the above-mentioned method is used. To efficiently obtain the desired metabolite, it is therefore necessary to study in detail the medium composition, the conditions for sterilizing the medium, the aeration-agitation rate, the pHs and temperatures of the medium and during the culturing of the highly productive strain, to determine and analyze various parameters relating to the culture, and to regulate the fermentation and metabolism based on the results.
Materials of the medium for culturing the microorganism are sometimes restricted so as to stabilize the fermentation conditions, etc., or acquire the desired substance economically. In order to achieve the efficient utilization of the medium materials as nutrients by the microorganism, however, it is necessary in some cases to impart novel genetic characters, which are necessary for altering the medium materials to substances the microorganism can utilize as nutrients, to the microorganism. In such a case, the impartment of these novel genetic characters by the usual mutagenesis treatment as described above is difficult. For these reasons, molecular breeding, which comprises transducing a specific alien gene into a specific microorganism by using genetic engineering techniques to thereby impart novel genetic characters to the microorganism, has been utilized. However, with respect to the microorganisms belonging to the class Hyphomycetes, which are different from bacteria, yeasts, etc., the transformation is frequently difficult, because, for example, there are known few autonomously replicating plasmids appropriate for the transformation of these microorganisms, many of which are polykaryocytes, and the frequencies of the protoplast generation and regeneration are extremely low. In fact, the transformation of microorganisms belonging to the class Hyphomycetes by genetic engineering techniques is practiced only by using fungi, for example, A. nidulans (see Proc. Natl. Acad. Sci., USA, 81, 1470(1984)), A. oryzae (see Agrc. Biol. Chem., 51, 323(1987)) or A. niger (see Curr. Genetics, 11, 499(1987)).
By the way, it has been known that some microorganisms belonging to the order Agonomycetales of the class Hyphomycetes produce useful substances, such as a cyclic depsipeptide having a vermifugal activity (i.e., substance PF102
REFERENCES:
patent: 5116815 (1992-05-01), Takagi et al.
Proc. Natl. Acad. Sci., USA, vol. 81, pp. 1470-1474, Mar. 1984: Transformation of Aspergillus nidulans by using a trpC plasmid by Yelton, Hamer and Timberlake.
Agric. Biol. Chem, 51 (2), pp. 323-328, 1987: Transformation of Aspergillus oryzea through Plasmid-mediated Complementation of the Methionine-auxotrophic Mutation by Iimura, Gomi, Uzu and Hara.
Curr Genet (1987) 11, pp. 499-503: Transformation of Aspergillus niger using the homologous orotidine-5'-phosphate-decarboxylase gene by Goosen, Bloemheuvel, Gysler, deBie, van den Broek & Swart.
Aoyagi Kaoru
Murakami Takeshi
Watanabe Manabu
Yanai Kohji
Lau Kawai
Meiji Seika Kaisha Ltd.
Patterson Jr. Charles L.
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