Microbial strains producing sphingolipid bases
AssigneeDsm N.V.
InventorsLex De Boer, Ingrid Francisca Caroline Van Der Wildt
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Abstract
Microbial strains capable of producing enhanced levels of sphingosine, dihydrosphingosine, phytosphingosine and/or derivatives thereof are disclosed. Additionally, there are disclosed methods based on mutagenesis, or other selection techniques, whereby such strains can be produced. As a preferred example thereof, mutant strains of Pichia are provided that are capable of producing about 50% or more of such compounds than wild type strains.
Claims18
A method for producing a mutant strain of yeast that is capable of producing at least 0.030 g tetraacetyl-phytosphingosine (TAPS)/g yeast dry weight comprising the steps of:
- a) obtaining a parent strain of yeast that is capable of naturally producing TAPS;
- b) treating said parent yeast strain with a mutagen;
- c) selecting a mutant strain of yeast, wherein said mutant strain is capable of producing at least 0.030 g TAPS/g yeast dry weight when cultured in liquid TAPS medium for a period of 3 days at 24° C. in a conical flask placed on an orbital incubator at 250 rpm.
A method for producing a mutant strain of yeast comprising the steps of:
- a) obtaining a parent strain of yeast that is capable of naturally producing TAPS;
- b) treating said parent yeast strain with a mutagen;
- and
- c) selecting a mutant strain of yeast that is capable of producing an amount of TAPS that is at least about 50% greater/g yeast dry weight than may be produced from said parent strain when compared under identical conditions, and wherein
- said mutant strain is also capable of producing at least 0.030 g TAPS/g yeast dry weight when cultured in liquid TAPS medium for a period of 3 days at 24° C. in a conical flask placed on an orbital incubator at 250 rpm.
A method for producing a compound selected from the group consisting of sphingosine, dihydrosphingosine, and phytosphingosine, or a derivative of any thereof, comprising the step of culturing in a liquid medium a composition, which composition includes a yeast strain capable of producing at least 0.030 g TAPS/g yeast dry weight when cultured free of other yeasts in liquid TAPS medium for a period of 3 days at 24° C. in a conical flask placed on an orbital incubator at 250 rpm, under conditions in which said compound is produced.
A method for producing a mutant strain of fungus that is capable of producing at least 0.030 g tetraacetyl-phytosphingosine (TAPS)/g yeast dry weight comprising the steps of:
- a) obtaining a parent strain of fungus that is capable of naturally producing TAPS;
- b) treating said parent fungus strain with a mutagen;
- c) selecting a mutant strain of fungus, wherein said mutant strain is capable of producing at least 0.030 g TAPS/g fungus dry weight when cultured in a suitable growth medium for a period of 3 days at 24° C.
A method for producing a mutant strain of bacteria that is capable of producing at least 0.030 g tetraacetyl-phytosphingosine (TAPS)/g bacteria dry weight comprising the steps of:
- a) obtaining a parent strain of bacteria that is capable of naturally producing TAPS;
- b) treating said parent bacterial strain with a mutagen;
- c) selecting a mutant strain of bacteria, wherein said mutant strain is capable of producing at least 0.030 g TAPS/g bacteria dry weight when cultured in a suitable growth medium for a period of 3 days at 24° C.
A composition comprising:
- (a) a Pichia strain, itself capable of producing at least 0.030 g TAPS/g Pichia dry weight when cultured in liquid TAPS medium for a period of 3 days at 24° C. in a conical flask placed on an orbital incubator at 250 rpm, maintained in contact with a growth medium under conditions that permit recovery of TAPS;
- and
- (b) at least one substance that permits extraction of said TAPS from said medium.
A method for producing a compound selected from the group consisting of ceramides, glycoceramides, and pseudoceramides, comprising the steps of:
- (a) culturing a Pichia strain in a liquid medium, which strain is capable of producing at least 0.030 g TAPS/g Pichia dry weight when cultured in liquid TAPS medium for a period of 3 days at 24° C. in a conical flask placed on an orbital incubator at 250 rpm, under conditions in which sphingosine, dihydrosphingosine, phytosphingosine, or a derivative of any thereof, is produced;
- (b) subjecting said product to at least one additional enzymatic or chemical step by which said ceramide, glycoceramide, or pseudoceramide is then produced;
- and
- (c) recovering said ceramide, glycoceramide, or pseudoceramide.
Dependencies are inferred from the claim text.