CA2256519A1 - Nutrient medium for increasing cell yield in fermentation - Google Patents
Nutrient medium for increasing cell yield in fermentation Download PDFInfo
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- CA2256519A1 CA2256519A1 CA002256519A CA2256519A CA2256519A1 CA 2256519 A1 CA2256519 A1 CA 2256519A1 CA 002256519 A CA002256519 A CA 002256519A CA 2256519 A CA2256519 A CA 2256519A CA 2256519 A1 CA2256519 A1 CA 2256519A1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
A nutrient medium used in fermentation for increasing the yield of cells or microorganism is provided. The formulation provided increases the yield of the fungus Lagenidium giganteum two-to three-fold over known media.
Description
CA 022~6~19 1998-09-ll NUTRIENT MEDIUM FOR INCREASING CELL YIELD IN FERMENTATION
FIELD OF THE INVENTION
This invention relates to a novel medium for use in fermentation which provides an 5 increased cell yield compared to that of known media. More particularly, the present invention produces at least a two to three-fold increase in the yield of the fungus Lagenidium giganteum compared to the yield obtained with known media. In addition to increasing yield of cells, L. giganteum grown in novel medium cont~ining lecithin exhibits increased effectiveness against mosquitoes.
BACKGROUND OF THE INVENTION
Fermentation is the process of growing microorg~ni~m~ or cells in specialized vessels. The cells or organisms may then be purified and used for a variety of purposes.
For instance, the fungus Lagenidium giganteum grown in fermenters is used as a 15 biocontrol agent for mosquitoes.
Optimal growth of the microorganism during fermentation depends on several factors including available nutrients, oxygen concentration, pH, temperature, and degree of mixing. Nutrients necessary for cell growth are provided in the medium used during the fermentation process. Accordingly, the yield obtained from fermentation depends, in part, 20 on the composition of the medium.
There are several published nutrient media currently used in the fermentation ofLagenidium giganteum. All use deionized water added to a final volume of 1 L, and all are sterilized. One formulation comprises 2.0 g ~rdamine pH, 2.0 g glucose, I mL corn oil, 0.5 g cholesterol and 2mM Ca2+. (Kerwin, James L. and Washino, Robert K. ( 1986)25 "Ground and aerial application of the sexual and asexual stages of Lagenidium giganteum (oomycetes: Lagenidiales) for mosquito control." J. Am. Mos. Control Assoc. 2(2): 182-1 89).
S~ 1 l I UTE SHEET (RULE 26) ... . .....
CA 022~6~19 1998-09-ll Another formulation comprises 2.0 g autolyzed yeast extract, 1.0 g proflo, 0.5 gfish meal, 2 mM CaCI22H20, lmM MgCI26H20, 0.05 g cholesterol and 2 mL cottonseedoil. (Kerwin, Jarnes L. and Washino, Robert K. (1988) "Field evaluation of Lagenidium giganteum (Oomycetes: Lagenidiales) and description of a natural epizootic involving a new isolate of fungus." J: Med Entomol. 2~(6): 452-460) Yet another ferrnentation medium comprises 1.25 g glucose, 1.25 g peptone, 1.25 g autolyzed yeast extract, 2 g corn oil, I g linseed oil, and 0.075 g CaCI22H20. (U.S. Patent No. 4,687,744). The fourth published medium contains 1.25 g yeast extract, 1.2 g glucose, 3.2 g powdered wheat germ, hemp seed extract to provide 250 mg/L of soluble protein, 1.25 g bactopeptone, 3 g glucose and 1.5 g corn oil. (Lord, Jeffrey C. and Roberts, Donald W. (1986) "The effects of culture medium quality and host passage on zoosporogensis and infectivity of Lagenidium giganteum (Oomycetes: Lagenidiales)," J. Invertebr. Pathol. 48:355-361) When used in ferrnentation, the above-referenced published medium formulations all yield approximately the sarne number of cells and infect susceptible mosquitoes at approximately the same rate. Thus, in order to increase the yield and infectivity of biocontrol agents like Lagenidium giganfeum, there is a need for an improved fermentation medium.
SUMMARY OF THE JNVFNTION
A medium for use in f~rment~tion consisting essentially of 3.6 g per liter peptone;
3.0 g per liter autolyzed yeast extract; 3.6 g per liter peptone; 1.5 to 3.0 g per liter autolyzed yeast extract; 1.6 g per liter cottonseed flour, such as ProFlo(~) (Traders Protein, Memphis, TN); 2.0 to 7.75 g per liter glucose (dextrose); 2.5 g per liter palm oil; 0.2 g per liter cholesterol; 0.6 g per liter CaC12 . 2H2O; 0.2 g per liter MgCl2. 6H20 and, optionally, 0.0 to 2.0 g per liter of lecithin. This medium provides increased yields of Lagenidium giganteum compared to prior art media, and, yield and infectivity of the organism is further increased when lecithin is included in the mediurn.
SUE~STlTUTE SHEET (RULE 26) CA 022~6~19 1998-09-11 DESCRIPTION OF THE PRFFERRFn FMP~ODIMF~TS
The present invention relates to an improved medium for fermentation. The medium increases yield at least approximately two to three fold over known media. The invention is useful in large scale production of Lagenidium giganteum, a biocontrol agent for mos4uitoes.
l~efinition~
As used herein, the term "ferrnentation" refers to the process of growing cells or microorg~ni~m.~ in specialized vessels. "Nutrient medium" ("mediurn") refers to a solid 10 or liquid substrate that will support the growth of an organism.
In a preferred embodiment of this invention, the nutrient medium is prepared as follows:
3.6 g per liter peptone;
1.5 to 3 g per liter autolyzed yeast extract;
15 1.6 g per liter cottonseed flour;
FIELD OF THE INVENTION
This invention relates to a novel medium for use in fermentation which provides an 5 increased cell yield compared to that of known media. More particularly, the present invention produces at least a two to three-fold increase in the yield of the fungus Lagenidium giganteum compared to the yield obtained with known media. In addition to increasing yield of cells, L. giganteum grown in novel medium cont~ining lecithin exhibits increased effectiveness against mosquitoes.
BACKGROUND OF THE INVENTION
Fermentation is the process of growing microorg~ni~m~ or cells in specialized vessels. The cells or organisms may then be purified and used for a variety of purposes.
For instance, the fungus Lagenidium giganteum grown in fermenters is used as a 15 biocontrol agent for mosquitoes.
Optimal growth of the microorganism during fermentation depends on several factors including available nutrients, oxygen concentration, pH, temperature, and degree of mixing. Nutrients necessary for cell growth are provided in the medium used during the fermentation process. Accordingly, the yield obtained from fermentation depends, in part, 20 on the composition of the medium.
There are several published nutrient media currently used in the fermentation ofLagenidium giganteum. All use deionized water added to a final volume of 1 L, and all are sterilized. One formulation comprises 2.0 g ~rdamine pH, 2.0 g glucose, I mL corn oil, 0.5 g cholesterol and 2mM Ca2+. (Kerwin, James L. and Washino, Robert K. ( 1986)25 "Ground and aerial application of the sexual and asexual stages of Lagenidium giganteum (oomycetes: Lagenidiales) for mosquito control." J. Am. Mos. Control Assoc. 2(2): 182-1 89).
S~ 1 l I UTE SHEET (RULE 26) ... . .....
CA 022~6~19 1998-09-ll Another formulation comprises 2.0 g autolyzed yeast extract, 1.0 g proflo, 0.5 gfish meal, 2 mM CaCI22H20, lmM MgCI26H20, 0.05 g cholesterol and 2 mL cottonseedoil. (Kerwin, Jarnes L. and Washino, Robert K. (1988) "Field evaluation of Lagenidium giganteum (Oomycetes: Lagenidiales) and description of a natural epizootic involving a new isolate of fungus." J: Med Entomol. 2~(6): 452-460) Yet another ferrnentation medium comprises 1.25 g glucose, 1.25 g peptone, 1.25 g autolyzed yeast extract, 2 g corn oil, I g linseed oil, and 0.075 g CaCI22H20. (U.S. Patent No. 4,687,744). The fourth published medium contains 1.25 g yeast extract, 1.2 g glucose, 3.2 g powdered wheat germ, hemp seed extract to provide 250 mg/L of soluble protein, 1.25 g bactopeptone, 3 g glucose and 1.5 g corn oil. (Lord, Jeffrey C. and Roberts, Donald W. (1986) "The effects of culture medium quality and host passage on zoosporogensis and infectivity of Lagenidium giganteum (Oomycetes: Lagenidiales)," J. Invertebr. Pathol. 48:355-361) When used in ferrnentation, the above-referenced published medium formulations all yield approximately the sarne number of cells and infect susceptible mosquitoes at approximately the same rate. Thus, in order to increase the yield and infectivity of biocontrol agents like Lagenidium giganfeum, there is a need for an improved fermentation medium.
SUMMARY OF THE JNVFNTION
A medium for use in f~rment~tion consisting essentially of 3.6 g per liter peptone;
3.0 g per liter autolyzed yeast extract; 3.6 g per liter peptone; 1.5 to 3.0 g per liter autolyzed yeast extract; 1.6 g per liter cottonseed flour, such as ProFlo(~) (Traders Protein, Memphis, TN); 2.0 to 7.75 g per liter glucose (dextrose); 2.5 g per liter palm oil; 0.2 g per liter cholesterol; 0.6 g per liter CaC12 . 2H2O; 0.2 g per liter MgCl2. 6H20 and, optionally, 0.0 to 2.0 g per liter of lecithin. This medium provides increased yields of Lagenidium giganteum compared to prior art media, and, yield and infectivity of the organism is further increased when lecithin is included in the mediurn.
SUE~STlTUTE SHEET (RULE 26) CA 022~6~19 1998-09-11 DESCRIPTION OF THE PRFFERRFn FMP~ODIMF~TS
The present invention relates to an improved medium for fermentation. The medium increases yield at least approximately two to three fold over known media. The invention is useful in large scale production of Lagenidium giganteum, a biocontrol agent for mos4uitoes.
l~efinition~
As used herein, the term "ferrnentation" refers to the process of growing cells or microorg~ni~m.~ in specialized vessels. "Nutrient medium" ("mediurn") refers to a solid 10 or liquid substrate that will support the growth of an organism.
In a preferred embodiment of this invention, the nutrient medium is prepared as follows:
3.6 g per liter peptone;
1.5 to 3 g per liter autolyzed yeast extract;
15 1.6 g per liter cottonseed flour;
2.0 to 7.75 g per liter glucose (dextrose);
2.5 g per liter palm oil;
0.2 g per liter cholesterol;
0.6 g per liter CaCl2 . 2H2O; and 20 0.2 g per liter MgCl2 . 6H2O.
Deionized water is added to a final volurne of 1 L and the pH is adjusted to 6.5.
The constituents are heated until dissolved and then the mediurn is sterilized by autoclaving at 121~C, 15 p.s.i., for 30 minutes. When used in the fennent~tion of 25 Lagenidium gigan~eum, this mediurn increases yield at least two to three fold over known media.
In another preferred embodiment, the nutrient mediurn is prepared by adding up to 2.0 g per liter of lecithin to the above forrnulation.
The following example is provided only for illustrative purposes, and is not to be 30 construed as limiting the invention in any way.
SU135TITUTE SHEET (RULE 26) CA 022~6~19 1998-09-11 Ex~rnple I
Sh~ke flask comparison of growth rates of Lagenidium giganteum in different media Growth rate in the novel nutrient medium was compared with two other media in side by side shake flask experiments.
Medium # 1:
1.25 g glucose (dextrose) 1.25 g peptone 1.25 g autolyzed yeast extract 2.0 g corn oil 1.0 g palm oil 0.03 g cholesterol 0.4 g CaC12 . 2H,O
0.2 g MgCI2 .6H20 Medium #2:
1.2 g peptone 1.2 g autolyzed yeast extract 3.0 g glucose (dextrose) 0.5 g cholesterol Novel Nutrient Medium:
3.6 g peptone 3.0 g autolyzed yeast extract 1.6 g Proflo cottonseed extract 2.0 g glucose (dextrose~
2.5 g palm oil 0.2 g cholesterol 0.6 g CaC12 ~ 2H2O
0.2 g MgC12 ~ 6H2O
When preparing each of the media, all ingredients were combined and deionized water was added to a final volume of 1 L. The pH was adjusted to 6.5. Contents were heated in a microwave until dissolved and then sterilized at 121 C~, 15 psi for 30 minlltes.
For each medium, nine 250 rnL flasks were each filled with 50 mL of medium. A disk of Lagenidium giganteum (California strain) taken from a petri dish was used to inoculate each flask. The flasks were shaken at 120 rpm, 29 C~ in an orbital temperature controlled SUBST~TUTE SHEET (RULE 2~i) CA 022~6~19 1998-09-11 s shaker for 7 days. Cells were harvested by centrifuging the fungal mass at 5,200 rpm for 20 minutes at 18 C~. The centrifuged cell mass was weighed and cell counts made with a hemacytometer. Mean cell counts were recorded. Results are summarized in Table 1.
Table 1 Medium #I Medium #2 Novel Nutrient Fold Increase Medium in cells/mL
when Novel Medium used Exp't# 1 1.2 - 2.0 x 10~ cells/mL 1.2-2.0 x I ob cells/mL 4.4 x I ob cells/mL 2.2 fold Exp't#2 6.25 x 10' cells/mL 7.5 x lO'cells/mL 1.38 x lObcells/mL 1.84-2.2 fold Exp't #3 2.97 x l O' cells/mL 3.3 x 10' cells/mL 4.75 x 10' cells/mL I .4- 1.6 fold Exp't#7 9.77x lO~cells/mL notdone 9.38x lO'cells/mL 9.6fold Exp't#8 1.93x lO'cells/mL notdone 7.30x lO~cells/mL 3.7fold Medium #1 and Medium #2 yielded approximately the same number of cells per mL of medium in each experiment. The novel nutrient mediurn consistently increased thenumber of cells/mL in comparison to either Medium #1 or Medium #2. The average yield of Lagenidium giganteum was increased approximately three and half fold when grown in the novel nutrient medium.
Fx~rr~le 2 Shake flask comparison of novel rnediurn with lecithin added Having established that the novel medium formulation of Example 1 increases cellyield over known media, the effect of varying amounts of dextrose and yeast extract and adding 1.0 g or 2.0 g lecithin to the basal novel medium was examined. All media were homogenized with a large probe at 70% speed for 10-15 seconds to ensure components were in solution. Using EmReagents color Phast(~), the pH of all media was adjusted to 6.5 and sterilized as in Example 1. For each medium, three 250 mL flasks were filled with 50 mL of medium, inoculated, cultured and harvested as described in Example 1. Results are summarized in Table 2 and Table 3 SUE~SmUTE SHEET (RULE 26) CA 022~6~19 1998-09-11 WO 98/58049 PCT~US97/10343 Table 2 Dextrose Yeast extract Lecithin Cell Yield % Wt % Wt % Wt (cells/mL) 0.8750 0.1250 0.0000 2.0 x 10~
0.8750 0.1250 0.0000 3.0 x 10~ Average Cell Yield (cells/mL) 0.6875 0.3125 0.0000 4.8x 10~ withoutlecithin:
3.6 x 105 0.5000 O.S000 0.0000 4.1 x 10 0-5000 0-5000 ~-~~~~ 4.13 x 0.5875 0.3125 0.1000 5.4 x 10 0.5875 0.3125 0.1000 4.05 x 10~
0.3000 0.5000 0.2000 7.4 x 105 Average Cell Yield (cells/mL) 0.3000 0.5000 0.2000 4.9 x 105 with lecithin:
2.5 g per liter palm oil;
0.2 g per liter cholesterol;
0.6 g per liter CaCl2 . 2H2O; and 20 0.2 g per liter MgCl2 . 6H2O.
Deionized water is added to a final volurne of 1 L and the pH is adjusted to 6.5.
The constituents are heated until dissolved and then the mediurn is sterilized by autoclaving at 121~C, 15 p.s.i., for 30 minutes. When used in the fennent~tion of 25 Lagenidium gigan~eum, this mediurn increases yield at least two to three fold over known media.
In another preferred embodiment, the nutrient mediurn is prepared by adding up to 2.0 g per liter of lecithin to the above forrnulation.
The following example is provided only for illustrative purposes, and is not to be 30 construed as limiting the invention in any way.
SU135TITUTE SHEET (RULE 26) CA 022~6~19 1998-09-11 Ex~rnple I
Sh~ke flask comparison of growth rates of Lagenidium giganteum in different media Growth rate in the novel nutrient medium was compared with two other media in side by side shake flask experiments.
Medium # 1:
1.25 g glucose (dextrose) 1.25 g peptone 1.25 g autolyzed yeast extract 2.0 g corn oil 1.0 g palm oil 0.03 g cholesterol 0.4 g CaC12 . 2H,O
0.2 g MgCI2 .6H20 Medium #2:
1.2 g peptone 1.2 g autolyzed yeast extract 3.0 g glucose (dextrose) 0.5 g cholesterol Novel Nutrient Medium:
3.6 g peptone 3.0 g autolyzed yeast extract 1.6 g Proflo cottonseed extract 2.0 g glucose (dextrose~
2.5 g palm oil 0.2 g cholesterol 0.6 g CaC12 ~ 2H2O
0.2 g MgC12 ~ 6H2O
When preparing each of the media, all ingredients were combined and deionized water was added to a final volume of 1 L. The pH was adjusted to 6.5. Contents were heated in a microwave until dissolved and then sterilized at 121 C~, 15 psi for 30 minlltes.
For each medium, nine 250 rnL flasks were each filled with 50 mL of medium. A disk of Lagenidium giganteum (California strain) taken from a petri dish was used to inoculate each flask. The flasks were shaken at 120 rpm, 29 C~ in an orbital temperature controlled SUBST~TUTE SHEET (RULE 2~i) CA 022~6~19 1998-09-11 s shaker for 7 days. Cells were harvested by centrifuging the fungal mass at 5,200 rpm for 20 minutes at 18 C~. The centrifuged cell mass was weighed and cell counts made with a hemacytometer. Mean cell counts were recorded. Results are summarized in Table 1.
Table 1 Medium #I Medium #2 Novel Nutrient Fold Increase Medium in cells/mL
when Novel Medium used Exp't# 1 1.2 - 2.0 x 10~ cells/mL 1.2-2.0 x I ob cells/mL 4.4 x I ob cells/mL 2.2 fold Exp't#2 6.25 x 10' cells/mL 7.5 x lO'cells/mL 1.38 x lObcells/mL 1.84-2.2 fold Exp't #3 2.97 x l O' cells/mL 3.3 x 10' cells/mL 4.75 x 10' cells/mL I .4- 1.6 fold Exp't#7 9.77x lO~cells/mL notdone 9.38x lO'cells/mL 9.6fold Exp't#8 1.93x lO'cells/mL notdone 7.30x lO~cells/mL 3.7fold Medium #1 and Medium #2 yielded approximately the same number of cells per mL of medium in each experiment. The novel nutrient mediurn consistently increased thenumber of cells/mL in comparison to either Medium #1 or Medium #2. The average yield of Lagenidium giganteum was increased approximately three and half fold when grown in the novel nutrient medium.
Fx~rr~le 2 Shake flask comparison of novel rnediurn with lecithin added Having established that the novel medium formulation of Example 1 increases cellyield over known media, the effect of varying amounts of dextrose and yeast extract and adding 1.0 g or 2.0 g lecithin to the basal novel medium was examined. All media were homogenized with a large probe at 70% speed for 10-15 seconds to ensure components were in solution. Using EmReagents color Phast(~), the pH of all media was adjusted to 6.5 and sterilized as in Example 1. For each medium, three 250 mL flasks were filled with 50 mL of medium, inoculated, cultured and harvested as described in Example 1. Results are summarized in Table 2 and Table 3 SUE~SmUTE SHEET (RULE 26) CA 022~6~19 1998-09-11 WO 98/58049 PCT~US97/10343 Table 2 Dextrose Yeast extract Lecithin Cell Yield % Wt % Wt % Wt (cells/mL) 0.8750 0.1250 0.0000 2.0 x 10~
0.8750 0.1250 0.0000 3.0 x 10~ Average Cell Yield (cells/mL) 0.6875 0.3125 0.0000 4.8x 10~ withoutlecithin:
3.6 x 105 0.5000 O.S000 0.0000 4.1 x 10 0-5000 0-5000 ~-~~~~ 4.13 x 0.5875 0.3125 0.1000 5.4 x 10 0.5875 0.3125 0.1000 4.05 x 10~
0.3000 0.5000 0.2000 7.4 x 105 Average Cell Yield (cells/mL) 0.3000 0.5000 0.2000 4.9 x 105 with lecithin:
4.63 x 105 0.6750 0.1250 0.2000 4.5 x 10 0.6750 0.1250 0.2000 4.1 x 10~
0.4875 0.3125 0.2000 3.6 x 10' As shown in Table 2, for media without lecithin, the average cells /mL yield is 3.6 x 105. With lecithin, yield increases to 4.63 x 105 cells/mL.
F,x~nlple 3 Infectivity of l ~genidium giganteum grown in various rnedia Lagenidium giganteum was grown in novel media described in Example 2 which contained no lecithin, 0.1000 % by weight lecithin or 0.2000 % by weight lecithin.
Culturing conditions were as described in Exarnple 1. The concentration of cells was calculated and their ability to kill mosquitoes measured at concentrations of 5,000; 2,500;
SUBSTITUTE SHEET (RULE 26) 1,250 and 675 cells/mL. Results summarized in Table 3 are averages of duplicate experiments.
Table 3 % Mortality at % Mortality at% Mortality at % Mortality at 5,000 cells/mL 2,500 cells/mL1,250 cells/mL 675 cells/mL
Medium without66 67 61 51 lecithin Medium with 87 87 89 74 lecithin These results illustrate that Lagenidium giganteum grown in the novel media killed more mos~uitoes than cells grown in media without added lecithin.
SU.,~ 111 UTE SHEET (RULE 26) . . .
0.4875 0.3125 0.2000 3.6 x 10' As shown in Table 2, for media without lecithin, the average cells /mL yield is 3.6 x 105. With lecithin, yield increases to 4.63 x 105 cells/mL.
F,x~nlple 3 Infectivity of l ~genidium giganteum grown in various rnedia Lagenidium giganteum was grown in novel media described in Example 2 which contained no lecithin, 0.1000 % by weight lecithin or 0.2000 % by weight lecithin.
Culturing conditions were as described in Exarnple 1. The concentration of cells was calculated and their ability to kill mosquitoes measured at concentrations of 5,000; 2,500;
SUBSTITUTE SHEET (RULE 26) 1,250 and 675 cells/mL. Results summarized in Table 3 are averages of duplicate experiments.
Table 3 % Mortality at % Mortality at% Mortality at % Mortality at 5,000 cells/mL 2,500 cells/mL1,250 cells/mL 675 cells/mL
Medium without66 67 61 51 lecithin Medium with 87 87 89 74 lecithin These results illustrate that Lagenidium giganteum grown in the novel media killed more mos~uitoes than cells grown in media without added lecithin.
SU.,~ 111 UTE SHEET (RULE 26) . . .
Claims (4)
1. A medium for use in fermentation, consisting essentially of:
(a) 3.6 g per liter peptone;
(b) 1.5 to 3 g per liter autolyzed yeast extract;
(c) 1.6 g per liter cottonseed flour;
(d) 2.0 to 7.75 g per liter glucose (dextrose);
(e) 2.5 g per liter palm oil;
(f) 0.2 g per liter cholesterol;
(g) 0.6 g per liter CaCl2 2H2O; and (h) 0.2 g per liter MgCl26H2O.
(a) 3.6 g per liter peptone;
(b) 1.5 to 3 g per liter autolyzed yeast extract;
(c) 1.6 g per liter cottonseed flour;
(d) 2.0 to 7.75 g per liter glucose (dextrose);
(e) 2.5 g per liter palm oil;
(f) 0.2 g per liter cholesterol;
(g) 0.6 g per liter CaCl2 2H2O; and (h) 0.2 g per liter MgCl26H2O.
2. The medium according to claim l, further comprising up to 2.0 g per liter lecithin.
3. The medium according to claim 1, for use in culturing Lagenidium giganteum.
4. The medium according to claim 2, for use in culturing Lagenidium giganteum.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/US1997/010343 WO1998058049A1 (en) | 1996-03-15 | 1997-06-17 | Nutrient medium for increasing cell yield in fermentation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2256519A1 true CA2256519A1 (en) | 1998-12-23 |
Family
ID=22261083
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002256519A Abandoned CA2256519A1 (en) | 1997-06-17 | 1997-06-17 | Nutrient medium for increasing cell yield in fermentation |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0954562A4 (en) |
| JP (1) | JP2002503090A (en) |
| KR (1) | KR19990087785A (en) |
| AU (1) | AU733497B2 (en) |
| CA (1) | CA2256519A1 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2015002678A (en) * | 2011-10-21 | 2015-01-08 | 株式会社カネカ | Culturing method of microorganisms, and method for producing pha by microorganisms |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| NL7704040A (en) * | 1976-04-28 | 1977-11-01 | Merck & Co Inc | PROCEDURE FOR PREPARING NEW ANTI-BIOTICS. |
| US4687744A (en) * | 1982-09-30 | 1987-08-18 | The Regents Of The University Of California | Artificial culture of the sexual stage of lagenidium giganteum |
| ATE142846T1 (en) * | 1991-01-10 | 1996-10-15 | Grace W R & Co | A PROCESS AND METHOD FOR THE PRODUCTION AND USE OF INSECT PATHOGENIC MUSHROOMS AS A PREPARATION FOR PEST CONTROL |
-
1997
- 1997-06-17 EP EP97930011A patent/EP0954562A4/en not_active Withdrawn
- 1997-06-17 KR KR1019980707266A patent/KR19990087785A/en not_active Application Discontinuation
- 1997-06-17 AU AU33942/97A patent/AU733497B2/en not_active Ceased
- 1997-06-17 JP JP53335898A patent/JP2002503090A/en active Pending
- 1997-06-17 CA CA002256519A patent/CA2256519A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| KR19990087785A (en) | 1999-12-27 |
| EP0954562A4 (en) | 2000-05-10 |
| EP0954562A1 (en) | 1999-11-10 |
| AU3394297A (en) | 1999-01-04 |
| JP2002503090A (en) | 2002-01-29 |
| AU733497B2 (en) | 2001-05-17 |
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