CN102660462B - Sterilization method of inorganic culture solution for different microalgae fermentation - Google Patents
Sterilization method of inorganic culture solution for different microalgae fermentation Download PDFInfo
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- CN102660462B CN102660462B CN 201210139238 CN201210139238A CN102660462B CN 102660462 B CN102660462 B CN 102660462B CN 201210139238 CN201210139238 CN 201210139238 CN 201210139238 A CN201210139238 A CN 201210139238A CN 102660462 B CN102660462 B CN 102660462B
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Abstract
The invention discloses a sterilization method of an inorganic culture solution for different microalgae fermentation, which belongs to the technical field of sterilization. The sterilization method comprises the steps of: adding sodium hypochlorite solid in the inorganic culture solution for the different microalgae fermentation, uniformly stirring the solution, standing, introducing sterile air into the solution to perform oxygen exposure and chloride dispersion, and then adding sodium thiosulfate in the solution to neutralize residual chloride; and adding the inorganic culture solution for the different microalgae fermentation and carbon and nitrogen nutrition sources in a sterilization tank for mother solution of the inorganic culture solution for the different microalgae fermentationto form the mother solution, performing routine sterile disinfection on the mother solution, sucking the inorganic culture solution for the different microalgae fermentation in a different microalgae fermentation device while adding the mother solution in the different microalgae fermentation device, and increasing the temperature in a main fermentation tank by excess heat of the mother solution, thereby finishing the process of sterilization and temperature rising of the inorganic culture solution for the different microalgae fermentation and then allowing the inoculation of algae seedlings, wherein the sterilization tank for the mother solution of the inorganic culture solution for the different microalgae fermentation is arranged at the external upper part of the different microalgae fermentation device.
Description
Technical field:
The invention belongs to the disinfecting technical field, relate to a kind of method for disinfection and sterilization of minimal media liquid, relate in particular to a kind of method for disinfection and sterilization of little algae heterotrophic fermentation minimal media liquid.
Background technology:
Splitting the kettle algae and claim again schizochytrium limacinum (Schizochytrium limacinum), is a kind of marine microalgae; Owing to can accumulate a large amount of greases in frond, can see many fat particles at microscopically, fat content reaches more than 50%, surpass some grease crop such as soybean, vegetable seed etc. at present, and these grease plant growth cycles need the several months long, not only be subjected to the restriction in season and area, and need take a large amount of soils; Only have 4-5 days and split kettle algae growth cycle, and be not subjected to the restriction in season, area and soil, the scale cultivation of splitting the kettle algae has become a kind of emerging biological industry; Splitting the kettle algae and be not only the potential raw material of biofuel and edible oil, is also the raw material of health care medicine industry; Contain a large amount of unsaturated fatty acidss in splitting the grease that the kettle algae produces, DHA(docosahexenoic acid particularly), be commonly called as DHA (docosahexaenoic acid), have brain healthy, impel child's brain and intelligence growth, delay cerebral atrophy, the prevention senile dementia, anticoagulation reduces the formation of thrombus, prevention myocardial infarction, cerebral infarction, reduce the sclerosis of blood fat prevention of arterial, the effects such as anti-inflammatory, anti-cancer; And unsaturated fatty acids is the nutrition of the biological young of marine fishery, and therefore splitting the kettle algae is good marine organisms bait; But, the heterotrophism cultivation technology before the kettle Cutleriales split is mostly according to microorganism fermentation process, and sterilizing process need carry out high-temperature steam sterilization to be forced coolingly again, not only needs a large amount of coal and water coolant, and need a large amount of additional facilities, as boiler, cooling tower etc.; Fermentation reactor is mostly pressurized vessel, adds corresponding environmental practice, and its capital expenditure and running cost are all higher; The present invention uses the sterilization of chemical process normal temperature, and nutrient chemical per ton only needs a small amount of sterilizing agent and energy consumption, and whole sterilization process has reduced Construction and operation cost and corresponding energy consumption, is a kind of low cost, high energy-conservation sterilization process.
Summary of the invention:
The object of the invention is to overcome the defective that exists in the method for disinfection and sterilization of existing little algae heterotrophic fermentation nutrient solution, seek to design the method for disinfection and sterilization of a kind of low cost, high energy-conservation little algae heterotrophic fermentation nutrient solution.
In order to realize the foregoing invention purpose, the present invention includes following steps:
(1) preparation of little algae heterotrophic fermentation minimal media liquid:
10-12 kilogram sodium-chlor, 7-10 kilo sulfuric acid magnesium, 3-4 kilogram potassium primary phosphate, 2-3 kilogram calcium chloride are dissolved in 1 ton of water stir, make little algae heterotrophic fermentation minimal media liquid;
(2) sterilization of little algae heterotrophic fermentation minimal media liquid:
(1) add solid clorox (NaClO in 1 ton of little algae heterotrophic fermentation minimal media liquid
3) the 2-5 gram, fully dissolving, stir, and standing 1-3 hour, again stirred 3-10 minute, standing more than 12 hours, make it give full play to the effect of sterilization;
(2) pass into the sterile air oxygen that exposes to the sun in little algae heterotrophic fermentation minimal media liquid and drove chlorine 20-40 minute, free chlorine is volatilized by aeration;
(3) oxygen that exposes to the sun drives and adds Sulfothiorine 5-10 gram after chlorine in little algae heterotrophic fermentation minimal media liquid, fully dissolving stirs evenly, with in fully and chlorine residue, with the chlorine residue in the little algae heterotrophic fermentation minimal media liquid of chlorine residue testing cassete test, testing cassete does not develop the color and represents that chlorine residue all is neutralized, by the Methylene blue vital staining, have or not the existence of viable bacteria with microscopic examination; Chlorine residue neutralization complete and nutrient solution in exist without viable bacteria can be with the little algae heterotrophic fermentation of the little algae heterotrophic fermentation minimal media liquid suction device after sterilization;
(4) little algae heterotrophic fermentation minimal media liquid mother liquor sterilization tank is set little algae heterotrophic fermentation device is outside upper, its volume is the 1/6-1/4 of little algae heterotrophic fermentation device;
(5) when giving little algae heterotrophic fermentation minimal media liquid disinfectant sterilization with clorox, add algae heterotrophic fermentation minimal media liquid and carbon and nitrogen nutrient sources in a subtle way in little algae heterotrophic fermentation minimal media liquid mother liquor sterilization tank, make little algae heterotrophic fermentation minimal media liquid mother liquor, and under 110-130 ℃ of 0.140-0.165Mpa pressure condition sterilization; The quality of little algae heterotrophic fermentation minimal media liquid is 4-6 times of carbon and nitrogen nutrient sources quality, and carbon and nitrogen nutrient sources is the known carbon and nitrogen nutrient sources that can be used in little algae cultivation of little algae heterotrophic fermentation technical field;
(6) to the suction of little algae heterotrophic fermentation device through the little algae heterotrophic fermentation minimal media liquid after the clorox sterilization and the little algae heterotrophic fermentation minimal media liquid mother liquor after sterilization under 110-130 ℃ of 0.140-0.165Mpa pressure condition, the waste heat of little algae heterotrophic fermentation minimal media liquid mother liquor promotes the temperature in little algae heterotrophic fermentation device, complete sterilization and the temperature-rise period of little algae heterotrophic fermentation minimal media liquid, can inoculate the algae kind.
The present invention compared with prior art adopts the sterilization of chemical process normal temperature, does not need special high temperature and cooling apparatus, saves cost and energy consumption; Utilize the waste heat of little algae heterotrophic fermentation minimal media liquid mother liquor to heat to little algae heterotrophic fermentation device, save special heating process, the energy-conservation amplitude of whole sterilizing process is large; Little algae heterotrophic fermentation device need not made pressurized vessel, saves cost of manufacture; Can be used in the sterilization of the sterilization that splits kettle algae heterotrophic fermentation minimal media liquid and little algae such as chlorella, spirulina pond cultivation minimal media liquid, little algae is cultivated had no adverse effects, the algae liquid of cultivating after the sterilization marine animal seedling of feeding, its growth, growth also have no adverse effects.
Embodiment:
The invention will be further described below by specific embodiment.
Embodiment 1:
The present embodiment comprises the following steps:
(1) preparation of little algae heterotrophic fermentation minimal media liquid:
11 kilograms of sodium-chlor, 8 kilo sulfuric acid magnesium, 3 kilograms of potassium primary phosphates, 2 kilograms of calcium chloride are dissolved in 1 ton of water and stir, make little algae heterotrophic fermentation minimal media liquid;
(2) sterilization of little algae heterotrophic fermentation minimal media liquid:
(1) add solid clorox (NaClO in 1 ton of little algae heterotrophic fermentation minimal media liquid
3) 3 grams, fully dissolving, stir, and standing 2 hours, again stirred 10 minutes, standing 15 hours, make it give full play to the effect of sterilization;
(2) pass into the sterile air oxygen that exposes to the sun in little algae heterotrophic fermentation minimal media liquid and drove chlorine 30 minutes, free chlorine is volatilized by aeration;
(3) oxygen that exposes to the sun drives and adds Sulfothiorine 8 grams after chlorine in little algae heterotrophic fermentation minimal media liquid, fully dissolving stirs evenly, with in fully and chlorine residue, with the chlorine residue in the little algae heterotrophic fermentation minimal media liquid of chlorine residue testing cassete test, testing cassete does not develop the color and represents that chlorine residue all is neutralized, by the Methylene blue vital staining, have or not the existence of viable bacteria with microscopic examination; Chlorine residue neutralization complete and nutrient solution in exist without viable bacteria can be with the little algae heterotrophic fermentation of the little algae heterotrophic fermentation minimal media liquid suction device after sterilization;
(4) little algae heterotrophic fermentation minimal media liquid mother liquor sterilization tank is set little algae heterotrophic fermentation device is outside upper, its volume is 1/5 of little algae heterotrophic fermentation device;
(5) when giving little algae heterotrophic fermentation minimal media liquid disinfectant sterilization with clorox, add algae heterotrophic fermentation minimal media liquid and carbon and nitrogen nutrient sources in a subtle way in little algae heterotrophic fermentation minimal media liquid mother liquor sterilization tank, make little algae heterotrophic fermentation minimal media liquid mother liquor, and under 120 ℃ of 0.150Mpa pressure conditions sterilization; The quality of little algae heterotrophic fermentation minimal media liquid is 5 times of carbon and nitrogen nutrient sources quality, and carbon and nitrogen nutrient sources is the known carbon and nitrogen nutrient sources that can be used in little algae cultivation of little algae heterotrophic fermentation technical field;
(6) to the suction of little algae heterotrophic fermentation device through the little algae heterotrophic fermentation minimal media liquid after the clorox sterilization and the little algae heterotrophic fermentation minimal media liquid mother liquor after sterilization under 120 ℃ of 0.150Mpa pressure conditions, the waste heat of little algae heterotrophic fermentation minimal media liquid mother liquor promotes the temperature in little algae heterotrophic fermentation device, complete sterilization and the temperature-rise period of little algae heterotrophic fermentation minimal media liquid, can inoculate the algae kind.
Claims (1)
1. the method for disinfection and sterilization of little algae heterotrophic fermentation minimal media liquid is characterized in that having the following steps:
(1) preparation of little algae heterotrophic fermentation minimal media liquid:
10-12 kilogram sodium-chlor, 7-10 kilo sulfuric acid magnesium, 3-4 kilogram potassium primary phosphate, 2-3 kilogram calcium chloride are dissolved in 1 ton of water stir, make little algae heterotrophic fermentation minimal media liquid;
(2) sterilization of little algae heterotrophic fermentation minimal media liquid:
(1) add solid clorox 2-5 gram in 1 ton of little algae heterotrophic fermentation minimal media liquid, fully dissolving, stir, and standing 1-3 hour, again stirred 3-10 minute, standing more than 12 hours, make it give full play to the effect of sterilization;
(2) pass into the sterile air oxygen that exposes to the sun in little algae heterotrophic fermentation minimal media liquid and drove chlorine 20-40 minute, free chlorine is volatilized by aeration;
(3) oxygen that exposes to the sun drives and adds Sulfothiorine 5-10 gram after chlorine in little algae heterotrophic fermentation minimal media liquid, fully dissolving stirs evenly, with in fully and chlorine residue, with the chlorine residue in the little algae heterotrophic fermentation minimal media liquid of chlorine residue testing cassete test, testing cassete does not develop the color and represents that chlorine residue all is neutralized, by the Methylene blue vital staining, have or not the existence of viable bacteria with microscopic examination; Chlorine residue neutralization complete and nutrient solution in exist without viable bacteria can be with the little algae heterotrophic fermentation of the little algae heterotrophic fermentation minimal media liquid suction device after sterilization;
(4) little algae heterotrophic fermentation minimal media liquid mother liquor sterilization tank is set little algae heterotrophic fermentation device is outside upper, its volume is the 1/6-1/4 of little algae heterotrophic fermentation device;
(5) when giving little algae heterotrophic fermentation minimal media liquid disinfectant sterilization with clorox, add algae heterotrophic fermentation minimal media liquid and carbon and nitrogen nutrient sources in a subtle way in little algae heterotrophic fermentation minimal media liquid mother liquor sterilization tank, make little algae heterotrophic fermentation minimal media liquid mother liquor, and under 110-130 ℃ of 0.140-0.165Mpa pressure condition sterilization; The quality of little algae heterotrophic fermentation minimal media liquid is 4-6 times of carbon and nitrogen nutrient sources quality, and carbon and nitrogen nutrient sources is the known carbon and nitrogen nutrient sources that can be used in little algae cultivation of little algae heterotrophic fermentation technical field;
(6) to the suction of little algae heterotrophic fermentation device through the little algae heterotrophic fermentation minimal media liquid after the clorox sterilization and the little algae heterotrophic fermentation minimal media liquid mother liquor after sterilization under 110-130 ℃ of 0.140-0.165Mpa pressure condition, the waste heat of little algae heterotrophic fermentation minimal media liquid mother liquor promotes the temperature in little algae heterotrophic fermentation device, complete sterilization and the temperature-rise period of little algae heterotrophic fermentation minimal media liquid, can inoculate the algae kind.
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| CN101831387A (en) * | 2010-04-16 | 2010-09-15 | 厦门大学 | Fully-synthetic culture medium, preparation method thereof and method for culturing Schizochytrium sp. by using same |
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| CN101831387A (en) * | 2010-04-16 | 2010-09-15 | 厦门大学 | Fully-synthetic culture medium, preparation method thereof and method for culturing Schizochytrium sp. by using same |
Non-Patent Citations (5)
| Title |
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| JP特开平9-173050A 1997.07.08 |
| 吴耀庭等.微藻异养化高细胞密度培养技术及其应用.《中国食品添加剂》.2008,第2008年卷(第6期),65-70. |
| 周文俊等.微藻异养化培养的研究进展.《海洋科学》.2012,第36卷(第2期), |
| 微藻异养化培养的研究进展;周文俊等;《海洋科学》;20120229;第36卷(第2期);136-142 * |
| 微藻异养化高细胞密度培养技术及其应用;吴耀庭等;《中国食品添加剂》;20081231;第2008年卷(第6期);65-70 * |
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Owner name: QINGDAO ZHONGREN ANIMAL PHARMACEUTICAL CO., LTD. Free format text: FORMER NAME: QINGDAO ZHONGREN PHARMACEUTICAL CO.LTD. |
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Address after: 266329, Qingdao, Shandong province Qingdao Jiaozhou West Industrial Park Road 6, China Ren Pharmaceutical Co., Ltd. Patentee after: QINGDAO ZHONGREN ANIMAL MEDICINE CO., LTD. Address before: 266329, Qingdao, Shandong province Qingdao Jiaozhou West Industrial Park Road 6, China Ren Pharmaceutical Co., Ltd. Patentee before: Qingdao Zhongren Pharmaceutical Co.Ltd. |