CN104357360B - A kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum and fermentation process - Google Patents

A kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum and fermentation process Download PDF

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CN104357360B
CN104357360B CN201410643050.1A CN201410643050A CN104357360B CN 104357360 B CN104357360 B CN 104357360B CN 201410643050 A CN201410643050 A CN 201410643050A CN 104357360 B CN104357360 B CN 104357360B
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addicted
liquid
feeding
raw material
sour micrococcus
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CN104357360A (en
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胡博旸
兰时乐
周芳如
肖调义
李玥
厉大伟
王红权
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Guangdong Chuangzhan Bona Agricultural Technology Co.,Ltd.
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Hunan Agricultural University
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    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor

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Abstract

A kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum and fermentation process, belong to microbial project fermentation technical field, this fermentation process is inoculated on MRS solid slant culture base by preservation addicted to sour micrococcus, and 37-40 DEG C of constant temperature culture 3-5d, obtains solid slope bacterial classification; To cultivate ripe solid slope strain inoculation in MRS liquid seed culture medium, 37-40 DEG C of constant temperature quiescent culture 24-36h, obtains liquid spawn; The inoculum size (V/V) of pressing 2-5% is again feeding addicted in sour micrococcus liquid raw material fermentation substratum to what prepare by cultured liquid-spawn inoculation, 37-40 DEG C of constant temperature quiescent culture 18-26h.Present method production technique is simple, low to the requirement of equipment, invest little, production cost is low, what obtain stablizes addicted to the sour micrococcus quality of the pharmaceutical preparations, Product Activity is high, be conducive to addicted to sour micrococcus liquid applying in animal farming industry, the animal product quality improving China had important practical significance, there is good economic benefit and social benefit simultaneously.

Description

A kind of feeding addicted to sour micrococcus liquid raw material fermentation substratum and fermentation process
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 (Pediococcusacidilactici), 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 ( pediococcusacidilactici) 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 (3)

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;
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.
2. 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 the inoculum size (V/V) of 2-5% by cultured liquid-spawn inoculation to according to claim 1 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.
3. as claimed in claim 2 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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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

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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

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