CN104357360A - Pediococcus acidilactici liquid raw material fermentation medium for feed and fermentation method - Google Patents
Pediococcus acidilactici liquid raw material fermentation medium for feed and fermentation method Download PDFInfo
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Abstract
The invention relates to a pediococcus acidilactici liquid raw material fermentation medium for feed and a fermentation method. The fermentation method comprises the following steps: inoculating a preserved pediococcus acidilactici to an MRS solid slant culture medium and carrying out constant-temperature incubation at 37-40 DEG C for 3-5 days to obtain a solid slant culture; inoculating the mature solid slant culture to an MRS liquid seed culture medium and carrying out constant temperature static culture at 37-40 DEG C for 24-36 hours to obtain a liquid culture; and then, inoculating the cultured liquid culture to the prepared pediococcus acidilactici liquid raw material fermentation medium for feed according to an inoculum size (V/V) of 2-5% and carrying out constant temperature static culture at 37-40 DEG C for 18-26 hours. The method is simple in production process, low in requirement on equipment, small in investment and low in production cost. The obtained pediococcus acidilactici preparation is stable in quality and high in activity, so that the pediococcus acidilactici liquid is applied to the animal breeding industry. The medium provided by the invention has important practical significance for improving the quality of animal products in China and also has good economic and social benefits.
Description
Technical field
The invention belongs to microbial project fermentation technical field, be specifically related to a kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum and fermentation process.
Background technology
Milk-acid bacteria is ratified the maximum class used and the probiotic bacterium used the earliest by various countries.This bacterium is widely used in cultured milk prod (as probiotics yogurt, probiotics fermention milky-drinks, probiotic bacterium cheese), fermented soybean milk products, functional food ingredient and foodstuff additive (interpolations etc. as in infant formula, person in middle and old age's milk powder, garden spgarden stuff etc.), dietary supplements (tablet, capsule etc.) and feed probiotics as the probiotics (or being called probiotic bacterium) of humans and animals.
Be a kind of milk-acid bacteria addicted to sour micrococcus (Pediococcus acidilactici), belong to gram-positive microorganism, do not move, amphimicrobian.At present, the soft tart flavour of food and fragrance can be given addicted to sour micrococcus, improve the quality of food and nutrition, also have antibacterial, reduce cholesterol, maintain micro-ecological environment, antitumor, resistance is different and the important biomolecule function such as strengthening immunity.It uses in multinational food as fermented bacterium is a large amount of, is the profitable strain enjoying people to like and accept.In addition, its significant fungistatic effect, makes to hide huge using value and wide application prospect addicted to sour micrococcus.
Addicted to sour micrococcus as a kind of novel probiotics, except the function with probiotics, also there is nourishing function, especially to people and the nontoxic not pathogenic of animal simultaneously, very strong resistibility is had to acid, can stomach be smoothly through, and breed rapidly in enteron aisle, reconcile intestinal microflora, secretion class sterilization factors, stimulating immune system improves immunological competence, and promoting animal full group's healthy water equality aspect, is the green additive in feed industry.
Raw material fermentation refers to that microorganism grows in the medium under high-temperature and high-pressure conditions, breeding and a kind of fermentation mode of metabolism.This fermentation method originates from abroad, is applied to wine industry the earliest, take zymamsis as point of penetration.To the later stage nineties 20th century, along with China's biotechnology flatly improves constantly, raw material fermentation is had new progress, technology is ripe gradually, and develops rapidly with other industry throughout the country very soon.Compared with cooked material fermentation, raw material fermentation decreases the nutritive loss in energy consumption and substratum.
At present about addicted to sour micrococcus raw material fermentation and formulation application few in the open source literature of animal cultivation, mainly concentrate on the fermentative production of pediocin and the application in sausage fermentation, as CN101050437 and CN102115720A individually discloses the method for producing pediocin; CN102127578A discloses a kind of method accumulating pediocin, can increase the turnout of pediocin; A strain sheet coccus disclosed in CN101289647 as the application of starter in fermented sausages, can improve the local flavor of fermented sausages; CN103283950A discloses the production method addicted to sour micrococcus active bacteria formulation (as additive for farm animal feed), but the method adopts traditional fermentation technique, the shortcoming such as have that production cost is high, complex process, facility investment are large.
Summary of the invention
Technical problem to be solved by this invention is: for above-mentioned the deficiencies in the prior art, there is provided a kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum and fermentation process, the method has that production technique is simple, facility investment is few, production cost and energy consumption low, be easy to the high and feature such as active high, three-waste free discharge of promotion and application, product viable count, solve at present addicted to sour micrococcus particularly addicted to the complex manufacturing that sour micrococcus production technique exists, the problems such as production cost is high, and facility investment is large.
In order to solve the problems of the technologies described above; the technical solution adopted in the present invention is: a kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum; this substratum is made up of following material by weight percentage: glucose 2-5%, peptone 0.5-0.8%, extractum carnis 0.3-0.5%, yeast extract paste 0.2-0.4%, anhydrous sodium acetate 0.8-1.2%, dipotassium hydrogen phosphate 0.1-0.2%, anhydrous magnesium sulfate 0.05-0.1%, thermal protecting agent 7-11%, miscellaneous bacteria inhibitor 0.1-3% and water 80-88%, pH5.5-7.0.
Above-mentioned thermal protecting agent is one or both in trehalose or glycerine.Above-mentioned miscellaneous bacteria inhibitor is one or both in sodium Diacetate or NaCl.
The preparation method of above-mentioned fermention medium: get glucose, peptone, extractum carnis, yeast extract paste, anhydrous sodium acetate, dipotassium hydrogen phosphate, anhydrous magnesium sulfate, thermal protecting agent and miscellaneous bacteria inhibitor in proportion and be added to the water; stir; glucose, peptone, extractum carnis, yeast extract paste, anhydrous sodium acetate, dipotassium hydrogen phosphate, anhydrous magnesium sulfate, thermal protecting agent and miscellaneous bacteria inhibitor are fully dissolved, then adjust ph is 5.5-7.0.PH value be adjusted to routine techniques, as adopt dilute NaOH solution, rare HCl solution regulate.
A kind of feeding addicted to sour micrococcus liquid raw material fermentation method, the method step is as follows:
(1) solid slope bacterial classification preparation: will be inoculated on fresh MRS solid slant culture base addicted to sour micrococcus slant strains (being preserved in 4 DEG C of environment), 37-40 DEG C of constant temperature culture 3-5d;
(2) liquid spawn preparation: by cultured solid slope bacterial classification, aseptically, picking one transfering loop cell is inoculated in through sterilizing (115-121 DEG C of sterilizing 25-30min) and is cooled in the MRS liquid seed culture medium of room temperature, 37-40 DEG C of constant temperature quiescent culture 24-36h;
(3) ferment: by the inoculum size (V/V) of 2-5% by cultured liquid-spawn inoculation to above-mentioned feeding addicted in sour micrococcus liquid raw material fermentation substratum, 37-40 DEG C of constant temperature quiescent culture 18-26h, obtains feeding addicted to sour micrococcus fermented liquid.
What use in the present invention is prior art addicted to sour micrococcus, and preserving number is: addicted to sour micrococcus (
pediococcus acidilactici) CGMCC1.4, CICC10344, CCAM080049 or CICIMB1355.
Compared with prior art, the present invention has the following advantages:
(1) what present method used directly adds in liquid raw material fermentation substratum addicted to sour micrococcus thermal protecting agent, to add addicted to sour micrococcus protectant adaptability, avoids protective material reduces thalline activity to the stimulation of thalline.
(2) the liquid raw material fermentation of present method employing, substratum does not need through high-temperature sterilization link, significantly reduces addicted to the energy consumption in sour micrococcus production process, it also avoid the loss of autoclaving process Middle nutrition material simultaneously.
(3) present method have that production technique is simple, facility investment is few, production cost is low, be easy to promotion and application, the high and feature such as active high, three-waste free discharge of product viable count.
Embodiment
Only the invention will be further described for following examples.
Embodiment 1
1, fermention medium preparation: get glucose 35kg, peptone 7kg, extractum carnis 3.5kg, yeast extract paste 3kg, anhydrous sodium acetate 10kg, dipotassium hydrogen phosphate 1.2kg, anhydrous magnesium sulfate 0.7kg, trehalose 85kg, NaCl21kg and sodium Diacetate 9kg and be placed in the container filling 824.6kg water, stir, each material is fully dissolved, and is 7.0 by dilute NaOH solution adjust ph.
2, solid slope bacterial classification preparation: be inoculated on fresh MRS solid slant culture base by what be preserved in 4 DEG C of refrigerators addicted to sour micrococcus slant strains, 37 DEG C of constant temperature culture 5d.
3, liquid spawn preparation: by cultured solid slope bacterial classification, aseptically, picking one transfering loop cell is inoculated in through sterilizing (121 DEG C of sterilizing 25min) and is cooled in the MRS liquid seed culture medium of room temperature, 40 DEG C of constant temperature quiescent culture 24h.
4, ferment: the inoculum size (V/V) by 4% is by cultured liquid-spawn inoculation in the aforementioned fermention medium prepared, and 40 DEG C of constant temperature quiescent culture 18h, obtain feeding addicted to sour micrococcus fermented liquid.
Embodiment 2
1, fermention medium preparation: get glucose 50kg, peptone 5kg, extractum carnis 3kg, yeast extract paste 4kg, anhydrous sodium acetate 8kg, dipotassium hydrogen phosphate 2kg, anhydrous magnesium sulfate 0.5kg, trehalose 60kg, glycerine 50kg, NaCl17.5kg be placed in the container filling 800kg water, stir, each material is fully dissolved, and is 5.5 by rare HCl solution adjust ph.
2, solid slope bacterial classification preparation: be inoculated on fresh MRS solid slant culture base by what be preserved in 4 DEG C of refrigerators addicted to sour micrococcus slant strains, 40 DEG C of constant temperature culture 3d.
3, liquid spawn preparation: by cultured solid slope bacterial classification, aseptically, picking one transfering loop cell is inoculated in through sterilizing (115 DEG C of sterilizing 30min) and is cooled in the MRS liquid seed culture medium of room temperature, 38 DEG C of constant temperature quiescent culture 30h.
4, ferment: the inoculum size (V/V) by 2% is by cultured liquid-spawn inoculation in the aforementioned fermention medium prepared, and 37 DEG C of constant temperature quiescent culture 26h, obtain feeding addicted to sour micrococcus fermented liquid.
Embodiment 3
1, fermention medium preparation: get glucose 20kg, peptone 8kg, extractum carnis 5kg, yeast extract paste 2kg, anhydrous sodium acetate 12kg, dipotassium hydrogen phosphate 1kg, anhydrous magnesium sulfate 1kg, glycerine 70kg and sodium Diacetate 1kg and be placed in the container filling 880kg water, stir, each material is fully dissolved, and is 6.0 by rare HCl solution adjust ph.
2, solid slope bacterial classification preparation: be inoculated on fresh MRS solid slant culture base by what be preserved in 4 DEG C of refrigerators addicted to sour micrococcus slant strains, 38 DEG C of constant temperature culture 4d.
3, liquid spawn preparation: by cultured solid slope bacterial classification, aseptically, picking one transfering loop cell is inoculated in through sterilizing (118 DEG C of sterilizing 28min) and is cooled in the MRS liquid seed culture medium of room temperature, 37 DEG C of constant temperature quiescent culture 36h.
4, ferment: the inoculum size (V/V) by 5% is by cultured liquid-spawn inoculation in the fermention medium of aforementioned preparation, and 39 DEG C of constant temperature quiescent culture 22h, obtain feeding addicted to sour micrococcus fermented liquid.
Claims (5)
1. one kind feeding addicted to sour micrococcus liquid raw material fermentation substratum; it is characterized in that; this substratum is made up of following material by weight percentage: glucose 2-5%, peptone 0.5-0.8%, extractum carnis 0.3-0.5%, yeast extract paste 0.2-0.4%, anhydrous sodium acetate 0.8-1.2%, dipotassium hydrogen phosphate 0.1-0.2%, anhydrous magnesium sulfate 0.05-0.1%, thermal protecting agent 7-11%, miscellaneous bacteria inhibitor 0.1-3% and water 80-88%, pH5.5-7.0.
2. as claimed in claim 1 a kind of feedingly to it is characterized in that addicted to sour micrococcus liquid raw material fermentation substratum, described thermal protecting agent is one or both in trehalose or glycerine.
3. as claimed in claim 1 a kind of feedingly to it is characterized in that addicted to sour micrococcus liquid raw material fermentation substratum, described miscellaneous bacteria inhibitor is one or both in sodium Diacetate or NaCl.
4. feeding addicted to a sour micrococcus liquid raw material fermentation method, it is characterized in that: the method step is as follows:
(1) solid slope bacterial classification preparation: will be inoculated on fresh MRS solid slant culture base addicted to sour micrococcus slant strains, 37-40 DEG C of constant temperature culture 3-5d;
(2) liquid spawn preparation: by cultured solid slope bacterial classification, aseptically, picking one transfering loop cell is inoculated in sterilized MRS liquid seed culture medium, 37-40 DEG C of constant temperature quiescent culture 24-36h;
(3) ferment: by feeding addicted in sour micrococcus liquid raw material fermentation substratum by according to any one of cultured liquid-spawn inoculation to claim 1-3 of the inoculum size (V/V) of 2-5%, 37-40 DEG C of constant temperature quiescent culture 18-26h, obtains feeding addicted to sour micrococcus fermented liquid.
5. as claimed in claim 4 a kind of feeding addicted to sour micrococcus liquid raw material fermentation method, it is characterized in that: the sterilizing in step (2) refers in 115-121 DEG C of sterilizing 25-30min, and is cooled to room temperature.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111504996A (en) * | 2020-05-14 | 2020-08-07 | 管维 | Test method of tea seed cake biological detoxification process |
Citations (4)
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|---|---|---|---|---|
| CN101497868A (en) * | 2009-02-26 | 2009-08-05 | 上海应用技术学院 | Improved MRS fluid nutrient medium and use method thereof for screening lactobacillus |
| CN101715815A (en) * | 2009-11-20 | 2010-06-02 | 润盈生物工程（上海）有限公司 | Method for producing ready-to-use leaven containing heat-resistant protective agents |
| CN101948783A (en) * | 2010-08-31 | 2011-01-19 | 广州广牧丰生物技术有限公司 | Culture medium capable of improving viable count of lactic acid bacillus |
| CN101971921A (en) * | 2010-11-12 | 2011-02-16 | 胡文锋 | Zinc-enriched inactivated lactobacillus microecological agent and preparation method thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101497868A (en) * | 2009-02-26 | 2009-08-05 | 上海应用技术学院 | Improved MRS fluid nutrient medium and use method thereof for screening lactobacillus |
| CN101715815A (en) * | 2009-11-20 | 2010-06-02 | 润盈生物工程（上海）有限公司 | Method for producing ready-to-use leaven containing heat-resistant protective agents |
| CN101948783A (en) * | 2010-08-31 | 2011-01-19 | 广州广牧丰生物技术有限公司 | Culture medium capable of improving viable count of lactic acid bacillus |
| CN101971921A (en) * | 2010-11-12 | 2011-02-16 | 胡文锋 | Zinc-enriched inactivated lactobacillus microecological agent and preparation method thereof |
Non-Patent Citations (2)
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| 吴琼: "乳链菌肽效价测定方法的研究", 《食品科学》, no. 06, 30 June 1999 (1999-06-30) * |
| 毛建伟等: "肉类发酵混合培养菌种的选择", 《食品科学》, no. 01, 30 January 1991 (1991-01-30) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111504996A (en) * | 2020-05-14 | 2020-08-07 | 管维 | Test method of tea seed cake biological detoxification process |
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Inventor after: Hu Boyang Inventor after: Lan Shile Inventor after: Zhou Fangru Inventor after: Xiao Diaoyi Inventor after: Li Yue Inventor after: Li Dawei Inventor after: Wang Hongquan Inventor before: Lan Shile Inventor before: Zhou Fangru Inventor before: Xiao Diaoyi Inventor before: Li Yue Inventor before: Li Dawei Inventor before: Wang Hongquan |
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Effective date of registration: 20201229 Address after: No.4 card on the first floor of Building 2, Yinjiang garden, Dawu Baimamiao District, Hongxing community, Nanjie street, Guangning County, Zhaoqing City, Guangdong Province, 526300 Patentee after: Guangdong Chuangzhan Bona Agricultural Technology Co.,Ltd. Address before: 410128 Furong district, Changsha, East Lake, Hunan, Hunan Agricultural University Patentee before: Hunan Agricultural University |
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