CN102199569B - Bacillus subtilis submerged fermentation culture medium applied to microbial fertilizers - Google Patents
Bacillus subtilis submerged fermentation culture medium applied to microbial fertilizers Download PDFInfo
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- CN102199569B CN102199569B CN 201110099582 CN201110099582A CN102199569B CN 102199569 B CN102199569 B CN 102199569B CN 201110099582 CN201110099582 CN 201110099582 CN 201110099582 A CN201110099582 A CN 201110099582A CN 102199569 B CN102199569 B CN 102199569B
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Abstract
The invention relates to a bacillus subtilis submerged fermentation culture medium for compound microbial fertilizers. The culture medium comprises the following components in percentage by weight: 0.8 to 1.5 percent of wheat powder, and 1.5 to 4 percent of bran powder. The composition of the culture medium is simple, the pH value is not required to be adjusted during the preparation, and the culture medium can promote the growth of thalli and the formation of spores, so that the quality of products is improved and the production cost is reduced.
Description
Technical field
The invention belongs to the microorganism culturing technical field, relate to the Technology that a kind of bacillus subtilis commerial scale liquid forms gemma, be specifically related to a kind of subtilis submerged fermentation culture medium that is applied to microbial fertilizer.
Background technology
Subtilis has vital role at aspects such as promotion plant growth, enhancing crop disease-resistant abilities, and is the microorganism of using safety, therefore is widely used in the microbial fertilizer production.
Because gemma has advantages of strong stress resistance, so the microorganism that contains in the microbial fertilizer exists mainly with the gemma form.Easily make bacillus subtilis spore with solid fermentation method, but labour intensity is large, and gemma concentration is not high, is generally 0.1 hundred million-1.0 hundred million CFU/g.And the agricultural industry criteria NY/T 798-2004(of People's Republic of China (PRC) composite microbiological fertilizer) in effective bacterial content of regulation pulvis and granulated fertilizer be not less than every gram 0.2 hundred million, every milliliter of effective bacterial content of liquid fertilizer is not less than 0.5 hundred million.Utilize the liquid submerged fermentation method to cultivate subtilis, cell concentration is high, but is difficult for forming gemma, and strictly the control culture condition could improve spore forming rate.Therefore the formation of gemma is subject to the impact of the each side such as pH, carbon source material concentration, nitrogen source concentration, dissolved oxygen, how to improve the gordian technique that gemma quantity in the fermented liquid just becomes fermentation of bacillus subtilis.
The substratum (take weight ratio) of general liquid culture bacillus subtilis spore is starch 2%, corn steep liquor 0.1%, dipotassium hydrogen phosphate 0.5%, ammonium sulfate 0.1%, calcium carbonate 0.25%, manganous sulfate 0.2-0.5%, pH 7.2-7.4.Culture condition: 37 ± 2 ℃, tank pressure 0.05MPa, air flow 1:1.0-1.2 volume/volume/minute, inoculum size 1-10%, fermentation 48-72h, spore forming rate 90%.The shortcoming of this fermention medium mainly is that the substratum preparation is comparatively complicated, and raw materials cost is higher.
Summary of the invention
The invention provides a kind of fermentation of bacillus subtilis substratum, improved culture medium prescription, reduce production costs.
The present invention is by the research discovery, and the ion that adds in traditional substratum can not need other interpolation by changing the substratum composition.
The main nutrient composition of fermentation of bacillus subtilis culture medium prescription of the present invention is wheat-flour, wheat bran, need not add other inorganic salt or trace element.
A kind of subtilis submerged fermentation culture medium for microbial fertilizer is characterized in that: by weight percentage, wheat-flour 0.8-1.5%, bran powder 1.5-4%, surplus is water.
The wheat-flour fineness is not less than 60 orders, and the bran powder fineness is not less than 40 orders.
Wheat bran is milling by product, and is cheap.But the direct abrasive dust of wheat-flour wheat, also red dog flour, substratum of the present invention does not contain soybean cake powder, and foam is less, needn't use defoamer, has reduced production cost.
After Medium's PH Value of the present invention is about 6.0(sterilization), subtilis is well-grown in the pH5.5-pH8.5 scope, so basal culture medium need not to use acid-alkali accommodation pH before sterilization, has simplified operation.
Carbon-nitrogen ratio in the substratum of the present invention is more reasonable, and not quite to stationary phase the time, pH is about 5.3-5.5, is conducive to the formation of gemma in the potential of hydrogen variation of whole fermenting process.
Adopt substratum of the present invention, make fermentation time from traditional 48-60 hour, shorten to 40-48 hour, the gemma concentration of 1m3 fermentor tank is 20-50 hundred million CFU/ml, has improved quality product, has reduced production cost.
Embodiment
Below by embodiment the present invention is done more detailed description, these embodiment only are the descriptions to best mode for carrying out the invention, and are not restricted to protection scope of the present invention.
Embodiment 1
Adopt (by weight) substratum of following proportioning: 8 kilograms of wheat-flours, 16 kilograms in wheat bran, water is settled to 0.8M3.After stirring, sterilized 25 minutes for 121 ℃, be cooled to 25-40 ℃, press the 5%(weight ratio) access subtilis seed liquor, fermentation condition: 37 ± 2 ℃, ventilation ratio 1:1.0 volume/volume/divide, rotating speed 200-250 rev/min, fermented 40 hours, residual sugar (By Anthrone Sulphuric acid method mensuration) is 5% before the inoculation, bacteria concentration (plate dilution assay method, parallel three times) average out to 38.5 hundred million CFU/ml, spore forming rate is about 90%.
Embodiment 2
Adopt (by weight) substratum of following proportioning: 12 kilograms of wheat-flours, 14 kilograms in wheat bran, water is settled to 0.8M
3After stirring, sterilized 25 minutes for 121 ℃, be cooled to 25-40 ℃, press the 5%(weight ratio) access subtilis seed liquor, fermentation condition: 37 ± 2 ℃, ventilation ratio 1:1.0 volume/volume/divide, rotating speed 280-320 rev/min, fermented 48 hours, residual sugar (By Anthrone Sulphuric acid method mensuration) is 8% before the inoculation, bacteria concentration (plate dilution assay method, parallel three times) average out to 42.4 hundred million CFU/ml, spore forming rate is about 95%.
Embodiment 3
Adopt (by weight) substratum of following proportioning: 7.2 kilograms of wheat-flours, 17 kilograms in wheat bran, water is settled to 0.8M3.After stirring, sterilized 25 minutes for 121 ℃, be cooled to 25-40 ℃, press the 5%(weight ratio) access subtilis seed liquor, fermentation condition: 37 ± 2 ℃, ventilation ratio 1:1.0 volume/volume/divide, rotating speed 300-350 rev/min, fermented 38 hours, residual sugar (By Anthrone Sulphuric acid method mensuration) is 3% before the inoculation, bacteria concentration (plate dilution assay method, parallel three times) average out to 45.6 hundred million CFU/ml, spore forming rate is about 90%.
Claims (2)
1. subtilis submerged fermentation culture medium that is used for microbial fertilizer is characterized in that: by weight percentage, wheat-flour 0.8-1.5%, bran powder 1.5-4%, surplus is water; The wheat-flour fineness is not less than 60 orders, and the bran powder fineness is not less than 40 orders.
2. substratum according to claim 1 is characterized in that: described Medium's PH Value is about 6.0 after the sterilization.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN 201110099582 CN102199569B (en) | 2011-04-21 | 2011-04-21 | Bacillus subtilis submerged fermentation culture medium applied to microbial fertilizers |
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| CN 201110099582 CN102199569B (en) | 2011-04-21 | 2011-04-21 | Bacillus subtilis submerged fermentation culture medium applied to microbial fertilizers |
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| CN102199569A CN102199569A (en) | 2011-09-28 |
| CN102199569B true CN102199569B (en) | 2013-01-16 |
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Families Citing this family (2)
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| CN102503601A (en) * | 2011-11-17 | 2012-06-20 | 大连春天生物技术有限公司 | Preparation method for bamboo fungus liquid strain culture medium |
| CN114516764A (en) * | 2021-08-30 | 2022-05-20 | 鹤壁市禾盛生物科技有限公司 | Method for preparing bacillus fermentation culture medium by using agricultural and sideline product waste |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321173C (en) * | 2004-12-02 | 2007-06-13 | 沈锦玉 | Bacillus subtilis preparation |
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| CN101107970B (en) * | 2007-08-23 | 2010-12-22 | 辽宁省微生物科学研究院 | Bion cow concentrated feed and method for making same |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321173C (en) * | 2004-12-02 | 2007-06-13 | 沈锦玉 | Bacillus subtilis preparation |
Non-Patent Citations (4)
| Title |
|---|
| Zhen-Min Chen 等.Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs.《Appl Microbiol Biotechnol》.2009,第85卷(第5期),1353–1360. * |
| 刘辉 等.一株枯草芽孢杆菌发酵培养基的优化.《山东农业科学》.2007,(第1期),100-101. * |
| 周映华 等.饲用枯草芽孢杆菌发酵条件的优化.《湖南农业科学》.2010,(第11期),21-23. * |
| 李超 等.枯草芽孢杆菌发酵培养基的优化.《饲料研究》.2009,(第9期),55-56. * |
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