CN110669678A - Griseofulvin strain solid culture medium - Google Patents
Griseofulvin strain solid culture medium Download PDFInfo
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- CN110669678A CN110669678A CN201911050270.2A CN201911050270A CN110669678A CN 110669678 A CN110669678 A CN 110669678A CN 201911050270 A CN201911050270 A CN 201911050270A CN 110669678 A CN110669678 A CN 110669678A
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- C12P17/04—Oxygen as only ring hetero atoms containing a five-membered hetero ring, e.g. griseofulvin, vitamin C
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Abstract
The invention discloses a griseofulvin strain solid culture medium which is prepared from the following raw materials in parts by weight: 100-150 parts of potato starch, 20-30 parts of cane sugar, 2-4 parts of monopotassium phosphate, 3-4 parts of potassium chloride, 1-2 parts of sodium nitrate, 0.4-0.5 part of magnesium sulfate, 0.01-0.02 part of ferrous sulfate and 15-25 parts of agar. The culture medium has smooth surface, moderate hardness and uniform inoculation; the bacterial colony cultured by the culture medium is full, white in color, round in shape, large in spore amount, good in spore synchronism and stable in bacterial fermentation unit; the culture medium is convenient to prepare, simple and easy to learn and high in working efficiency.
Description
Technical Field
The invention belongs to the technical field of strain culture media, and particularly relates to a griseofulvin strain solid culture medium.
Background
Griseofulvin (griseofulvin) belongs to aromatic derivative antibiotics, and is an antifungal antibiotic. Griseofulvin is widely used in clinical treatment of fungal infections of skin and stratum corneum; has good application prospect in the aspect of agricultural plant protection. The strain used for producing the griseofulvin through domestic fermentation is a descendant N15-3-40 of a D-756 strain obtained by artificially mutagenizing a wild type Penicillium patulum 4541 strain which is self-screened in China.
At present, the traditional sucrose formula culture medium and PDA solid culture medium are used for preparing the industrial production strain of the griseofulvin of domestic enterprises. The sucrose formula culture medium is prepared from the following raw materials in percentage by weight: 3% of sucrose, 0.5% of monopotassium phosphate, 0.4% of potassium chloride, 0.2% of sodium nitrate, 0.05% of magnesium sulfate, 0.001% of ferrous sulfate and 2.5% of agar. The PDA solid culture medium is prepared from the following raw materials in percentage by weight: 20% of potatoes, 2% of glucose and 15-20% of agar powder. The quality of the culture medium is one of the key factors in the preparation process of griseofulvin strains. The traditional sucrose formula culture medium has long culture period and unstable hardness; the quality of the component potatoes in the traditional PDA culture medium formula is greatly different due to different varieties, seasons, production places and other factors, and the stability of griseofulvin strains can also be influenced.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a griseofulvin strain solid culture medium, which has the advantages of stable strain quality, short strain culture period and high strain fermentation unit.
The technical scheme of the invention is realized as follows: a griseofulvin strain solid culture medium is prepared from the following raw materials in parts by weight: 100-150 parts of potato starch, 20-30 parts of cane sugar, 2-4 parts of monopotassium phosphate, 3-4 parts of potassium chloride, 1-2 parts of sodium nitrate, 0.4-0.5 part of magnesium sulfate, 0.01-0.02 part of ferrous sulfate and 15-25 parts of agar.
The technical scheme is further optimized, and the griseofulvin strain solid culture medium is prepared from the following raw materials in parts by weight: 120 parts of potato starch, 20 parts of cane sugar, 4 parts of monopotassium phosphate, 3 parts of potassium chloride, 1 part of sodium nitrate, 0.4 part of magnesium sulfate, 0.02 part of ferrous sulfate and 20 parts of agar.
The preparation method of the griseofulvin strain solid culture medium comprises the following steps: weighing the components in proportion, fully dissolving the components in deionized water, fixing the volume to a target volume, finally adjusting the pH to 5-6, preferably adjusting the pH to 5.5, subpackaging, sterilizing by using a sterilization kettle at the temperature of 115-125 ℃ for 20-30 minutes, and pouring a flat plate or placing an inclined plane in a sterile room when the temperature is reduced to about 60 ℃.
Compared with the prior art, the invention has the advantages that:
potatoes are not used as a raw material of a culture medium, so that the influence of unstable factors of potato varieties, seasons and production places is overcome; the culture period is shortened from 7 days to 5 days; the culture medium has smooth surface, moderate hardness and uniform inoculation; the bacterial colony cultured by the culture medium is full, white in color, round in shape, large in spore amount, good in spore synchronism and stable in bacterial fermentation unit; the culture medium is convenient to prepare, simple and easy to learn and high in working efficiency.
Drawings
FIG. 1 the appearance of the mature slant lawn colonies of the invention and two conventional strains;
in fig. 1: a is the appearance of the bacterial colony of the culture medium bacterial strain mature inclined plane, B is the appearance of the bacterial colony of the culture medium bacterial strain mature inclined plane of the sucrose formula, and C is the appearance of the bacterial colony of the bacterial strain mature inclined plane of the PDA solid culture medium.
FIG. 2 shows the case of microscopic examination of the mature slant of the strain of the present invention and two conventional strains;
in fig. 2: d, the culture medium slant spores of the invention, E is the culture medium slant spores of the sucrose formula, and F is the slant spores of the PDA solid culture medium.
Detailed Description
In order to make the technical solution of the present invention clearer, the present invention is further described in detail by specific examples and comparative experiments below.
Example 1: a griseofulvin strain solid culture medium is prepared from the following raw materials in parts by weight: 120 parts of potato starch, 20 parts of cane sugar, 4 parts of monopotassium phosphate, 3 parts of potassium chloride, 1 part of sodium nitrate, 0.4 part of magnesium sulfate, 0.02 part of ferrous sulfate and 20 parts of agar, and the pH value is adjusted to 5.5.
Example 2: a griseofulvin strain solid culture medium is prepared from the following raw materials in parts by weight: 125 parts of potato starch, 25 parts of cane sugar, 3 parts of monopotassium phosphate, 4 parts of potassium chloride, 2 parts of sodium nitrate, 0.5 part of magnesium sulfate, 0.01 part of ferrous sulfate and 18 parts of agar, and adjusting the pH value to 5.3.
Example 3: a griseofulvin strain solid culture medium is prepared from the following raw materials in parts by weight: 140 parts of potato starch, 30 parts of cane sugar, 3.5 parts of monopotassium phosphate, 3.5 parts of potassium chloride, 1.5 parts of sodium nitrate, 0.45 part of magnesium sulfate, 0.02 part of ferrous sulfate and 22 parts of agar, and the pH value is adjusted to 5.7.
And (3) comparison test:
1. preparation of the culture medium of the invention:
the griseofulvin strain solid culture medium of the invention is prepared according to the formula proportion of the example 1:
1) weighing 120g of potato starch, 20g of sucrose, 4g of monopotassium phosphate, 3g of potassium chloride, 1g of sodium nitrate, 0.4g of magnesium sulfate and 0.02g of ferrous sulfate according to the formula proportion of example 1, and adding the weighed materials into a 2000ml beaker;
2) adding deionized water into a beaker, fully dissolving all components in the beaker, and fixing the volume to 1000 ml;
3) dripping 10% NaOH solution into a beaker, and adjusting the pH value to 5.5 to prepare a culture medium nutrient solution;
4) the method for inverting the plate comprises the following steps: weighing 4g of agar in 500ml of conical flasks, weighing 200ml of culture medium nutrient solution, subpackaging in each conical flask, plugging a cotton plug, and bundling by using kraft paper;
the eggplant bottle slope method comprises the following steps: adding 1g of agar into each eggplant bottle, measuring 50ml of culture medium nutrient solution, subpackaging in each eggplant bottle, plugging a cotton plug, and bundling with kraft paper;
5) sterilizing at 121 deg.C for 25 min, and pouring plate or placing inclined plane in sterile room when the temperature is reduced to about 60 deg.C.
2. Preparing a sucrose formula culture medium:
1) weighing 30g of sucrose, 5g of monopotassium phosphate, 4g of potassium chloride, 2g of sodium nitrate, 0.5g of magnesium sulfate and 0.01g of ferrous sulfate according to the proportion of a traditional formula, and adding into a 2000ml beaker;
2) adding deionized water into a beaker, fully dissolving all components in the beaker, and then fixing the volume to 1000ml to prepare a culture medium nutrient solution;
3) the method for inverting the plate comprises the following steps: weighing 4g of agar in 500ml of conical flasks, weighing 200ml of culture medium nutrient solution, subpackaging in each conical flask, plugging a cotton plug, and bundling by using kraft paper;
the eggplant bottle slope method comprises the following steps: adding 1g of agar into each eggplant bottle, measuring 50ml of culture medium nutrient solution, subpackaging in each eggplant bottle, plugging a cotton plug, and bundling with kraft paper;
4) sterilizing at 121 deg.C for 25 min, and pouring plate or placing inclined plane in sterile room when the temperature is reduced to about 60 deg.C.
3. Preparing a PDA solid culture medium:
1) peeling fresh potato, cutting into 1cm square pieces, weighing 200g, adding 1000ml water, boiling, maintaining for 30min, filtering with 2 layers of gauze in 2000ml beaker, removing residue, and collecting filtrate;
2) weighing 20g of glucose, adding into the filtrate, fully dissolving, and metering to 1000ml to obtain a culture medium nutrient solution;
3) the method for inverting the plate comprises the following steps: weighing 4g of agar in 500ml of conical flasks, weighing 200ml of culture medium nutrient solution, subpackaging in each conical flask, plugging a cotton plug, and bundling by using kraft paper;
the eggplant bottle slope method comprises the following steps: adding 1g of agar into each eggplant bottle, measuring 50ml of culture medium nutrient solution, subpackaging in each eggplant bottle, plugging a cotton plug, and bundling with kraft paper;
4) sterilizing at 121 deg.C for 25 min, and pouring plate or placing inclined plane in sterile room when the temperature is reduced to about 60 deg.C.
4. Spore counting and fermentation validation
Griseofulvin strains are respectively inoculated on the inclined planes of eggplant bottles of the culture medium, the sucrose formula culture medium and the PDA solid culture medium, the inoculation amount of each inclined plane is consistent, after inoculation, the culture is carried out at 28 +/-0.5 ℃, and after the culture is mature, the spore count and the comparative verification test result of a fermentation unit of the three culture media are shown in table 1.
TABLE 1 spore count and fermentation Unit validation comparison data
| Culture medium | Period of time | Spore count (one) | Fermentation Unit mu/ml |
| The culture medium of the invention | 5 days | 3.27×108 | 21712 |
| Sucrose formula culture medium | 7 days | 2.4×108 | 20873 |
| PDA solid culture medium | 5 days | 1.9×108 | 20563 |
From the test results, the slant strain spore amount and the fermentation unit cultured by the culture medium have certain advantages, the culture period is short, and the advantages of two traditional culture media are considered.
5. Microscopic observation of spores
Griseofulvin strains are respectively inoculated on the inclined planes of eggplant bottles of the culture medium, the sucrose formula culture medium and the PDA solid culture medium, the inoculation amount of each inclined plane is ensured to be consistent, after inoculation, the culture is carried out at 28 +/-0.5 ℃, after the culture is mature, the inclined plane bacterial colony appearances of the three culture media are observed, and the spore morphology and the growth synchronism are observed through microscopic examination. The appearance of the mature slant lawn bacterial colony is shown in figure 1, and the microscopic examination condition is shown in figure 2.
From the appearances of three kinds of mature slant lawn bacterial colonies, the slant lawn of the culture medium is whiter, and a single bacterial colony is full and uniform in size. From the condition of spore microscopic examination, the shapes of the spores on the three slopes are relatively uniform, and the synchronism is good. Therefore, in summary, the culture medium is suitable for griseofulvin strains and can replace the traditional culture medium.
Claims (4)
1. A griseofulvin strain solid culture medium is prepared from the following raw materials in parts by weight: 100-150 parts of potato starch, 20-30 parts of cane sugar, 2-4 parts of monopotassium phosphate, 3-4 parts of potassium chloride, 1-2 parts of sodium nitrate, 0.4-0.5 part of magnesium sulfate, 0.01-0.02 part of ferrous sulfate and 15-25 parts of agar.
2. The griseofulvin strain solid culture medium according to claim 1, characterized in that: the paint is prepared from the following raw materials in parts by weight: 120 parts of potato starch, 20 parts of cane sugar, 4 parts of monopotassium phosphate, 3 parts of potassium chloride, 1 part of sodium nitrate, 0.4 part of magnesium sulfate, 0.02 part of ferrous sulfate and 20 parts of agar.
3. The griseofulvin strain solid culture medium according to claim 1 or 2, characterized in that: the preparation method comprises the following steps: weighing the components in proportion, fully dissolving the components in deionized water, fixing the volume to a target volume, finally adjusting the pH value to 5-6, subpackaging the components, sterilizing the components for 20-30 minutes at the temperature of 115-125 ℃ by using a sterilization kettle, and pouring a flat plate or swinging an inclined plane in a sterile room when the temperature is reduced to about 60 ℃.
4. The griseofulvin strain solid culture medium according to claim 3, characterized in that: the pH was adjusted to 5.5.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112080438A (en) * | 2020-10-15 | 2020-12-15 | 内蒙古格林特制药有限责任公司 | Griseofulvin low-foam-yield strain and preparation method and application of low-foam-yield strain |
| CN112094874A (en) * | 2020-09-16 | 2020-12-18 | 内蒙古格林特制药有限责任公司 | Culture medium for producing griseofulvin through fermentation |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101812489A (en) * | 2010-04-29 | 2010-08-25 | 赤峰制药股份有限公司 | Culture medium for producing griseofulvin by fermentation |
| CN104012298A (en) * | 2014-04-29 | 2014-09-03 | 潢川九龙春天农业科技有限公司 | Edible fungus liquid seed submerged fermentation packaged seed production technology and medium formulas thereof |
| CN107267398A (en) * | 2017-06-16 | 2017-10-20 | 江阴市长泾国民育种场 | A kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method |
| CN108841733A (en) * | 2018-07-26 | 2018-11-20 | 福州工微生物科技有限公司 | The one plant of production main part of Songgangmeisu --- bacterial strain and method of griseofulvin |
-
2019
- 2019-10-22 CN CN201911050270.2A patent/CN110669678A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101812489A (en) * | 2010-04-29 | 2010-08-25 | 赤峰制药股份有限公司 | Culture medium for producing griseofulvin by fermentation |
| CN104012298A (en) * | 2014-04-29 | 2014-09-03 | 潢川九龙春天农业科技有限公司 | Edible fungus liquid seed submerged fermentation packaged seed production technology and medium formulas thereof |
| CN107267398A (en) * | 2017-06-16 | 2017-10-20 | 江阴市长泾国民育种场 | A kind of Liquid Strain of Ganoderma Lucidum culture medium prescription and Liquid Strain of Ganoderma Lucidum cultural method |
| CN108841733A (en) * | 2018-07-26 | 2018-11-20 | 福州工微生物科技有限公司 | The one plant of production main part of Songgangmeisu --- bacterial strain and method of griseofulvin |
Non-Patent Citations (1)
| Title |
|---|
| 董海龙等: "主要营养和环境因素对疣孢漆斑菌X-16菌株产孢的影响", 《山西农业科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112094874A (en) * | 2020-09-16 | 2020-12-18 | 内蒙古格林特制药有限责任公司 | Culture medium for producing griseofulvin through fermentation |
| CN112080438A (en) * | 2020-10-15 | 2020-12-15 | 内蒙古格林特制药有限责任公司 | Griseofulvin low-foam-yield strain and preparation method and application of low-foam-yield strain |
| CN112080438B (en) * | 2020-10-15 | 2023-05-12 | 内蒙古格林特制药有限责任公司 | Preparation method and application of griseofulvin low-foam-production strain and low-foam-production strain |
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