CN1560231A - Preparation process of fermenting agent for beneficial growing lactic bacteria and application in fresh cheese thereof - Google Patents
Preparation process of fermenting agent for beneficial growing lactic bacteria and application in fresh cheese thereof Download PDFInfo
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Abstract
The invention is a method of preparing probiotic lactobacillus starter and its application in fresh cheeses. The method includes the steps: culture medium preparing, strain culturing, concentrating, adding protective agent, drying, etc. It uses one or many ones of the defatted milk, whey, lactose and physiological saline as activating liquor, activates the dry solid starter by the activating liquor, then makes the activated starter enter in the routine production of fermented milk products like acidophilus milk, cheeses, etc by using bacteriophage-free fresh milk or recovered milk as raw material, and makes the probiotic bacteria-containing fermented milk products. It makes probiotic lactobacillus into high-activity solid preparation, reduces the working procedure of amplified cultivation of the starter in the course of producing fermented milk products, and the product quality more stable.
Description
[technical field]: the present invention relates to a kind of microbial preparation, the preparation of particularly solid-state high reactivity probiotic lactobacillus starter and corresponding fresh cheese product, and contain application in the fresh cheese goods of lactobacterium casei viable bacteria in processing.
[background technology]: dairy products are the most approaching perfect food, almost contain the required whole nutritive ingredients of biological activity, are the important component that constitutes balance diet.Fermented-milk is meant and adds starter in fluid milk, fermentative action by microorganism is converted into lactic acid with lactose, has reduced the pH value and the acidity of milk-product, this has not only given milk-product special mouthfeel and quality, also reduces or has avoided the eater to produce the possibility of lactose intolerance reaction.Fresh cheese is a kind of of fermented-milk, is through lactic fermentation and without a sophisticated class cheese product.During the fermentation, because action of microorganisms, protein of milk progressively is broken down into peptide, peptone, amino acid etc., and these materials are easy to be absorbed by the body, and makes protein digestibility in the fermented-milk up to 96%～98%; A large amount of indispensable amino acids and bio-active substance are verified and are promoted the health of body to produce positive effect.
The difference of lactobacillus starter not only affects the process parameter in the cultured milk prod production, also directly affects taste, quality and the nutritive value of finished product.Conventional ferment agent for sour milk bacterial classification is lactobacillus bulgaricus and thermophilus streptococcus, and they can finish the effect of fermentation and acid in milk-product, but they are foreign matters for the normal microflora in the enteron aisle, can not grow surely in enteron aisle.In addition, with regard to the use-pattern of conventional lactobacillus starter, mostly be repeatedly going down to posterity of liquid bacterial classification, need take certain fund, equipment and personnel, and pollute easily and degeneration in the process that goes down to posterity, cause the instability of product quality.Chinese patent CN1105521A has announced a kind of preparation method of dry acid milk powder, the dry acid milk powder that this method is produced can be used as the starter of conventional milk-acid bacteria, but, wherein only contain conventional yoghurt bacterium, and the bacterium vigor is 10 because bacterial classification that adopts and complete processing is different
7/ gram.
With regard to probiotic bacterium, people had research to the application of bifidus bacillus in fermented-milk, had announced a kind of high method for preparing sour milk that contains bifidus bacillus as Chinese patent CN1240586A.Though it is the probiotics in the human intestinal that the deficiency of this patent is the bacterial classification bifidus bacillus as starter, it only is 10 that physiological properties such as itself anaerobism cause the probiotic bacterium number in the finished product
6/ gram is difficult to guarantee that it keeps higher biological activity in cultured milk prod and human body alimentary canal.
[summary of the invention]: the object of the invention is exactly to solve conventional ferment agent for sour milk bacterial classification can not grow surely in enteron aisle, its use need repeatedly be gone down to posterity, take certain fund, equipment and personnel, and pollute easily and degeneration in the process that goes down to posterity, cause the problems such as instability of product quality, a kind of preparation method of probiotic lactobacillus starter and the application in fresh cheese thereof are provided.
The preparation method of probiotic lactobacillus starter provided by the invention, realize by following steps:
---a culture medium preparation: this substratum is solvent with the deionized water, and volume ratio g/L adds following ingredients successively by weight: peptone 8～10, yeast extract paste 3～5, extractum carnis 1～3, glucose 4～6, sodium acetate 2～6, tween 80 1～2, ferric sulfate 1～3, manganous sulfate 1～2, triammonium citrate 1～2;
In the above-mentioned substratum configuration, also volume ratio g/L adds following material by weight: magnesium sulfate heptahydrate 1～3, dipotassium hydrogen phosphate 1～3, Trisodium Citrate 2～5, four water manganous sulfates 2～4.
---b spawn culture: with lactobacterium casei, add in the above-mentioned substratum by inoculum size 2～5%, culture temperature is 30～37 ℃, in the culturing process by weight volume ratio stream add carbon source 3-7g/L and/or nitrogenous source 3-5g/L, the pH value of control substratum is 5.0～6.5, incubation time 5～10 hours reaches 10 to the bacterium number
9～10/ mL obtains the high-concentration culturing thing of lactobacterium casei;
It is glucose, lactose, semi-lactosi, sucrose, maltose that described stream adds carbon source;
It is whey, skimming milk, casein, soybean protein, amino acid that described stream adds nitrogenous source;
Volume ratio stream is with Radix Dauci Sativae juice and/or each 10～20g/L of tomato juice, to promote the growth of thalline by weight.
---c concentrates: above-mentioned high-concentration culturing thing is concentrated through rotating speed 3000～5000rpm/10～25min is centrifugal, obtain the enriched material of active lactobacillus casei;
---d adds protective material: in the enriched material of above-mentioned lactobacterium casei, add protective material, protectant consumption be in b step centrifugal before high-concentration culturing liquid long-pending 10～20%, make bacteria suspension;
Described protective material is one or more in sucrose, pectin, skimming milk, whey, serum albumin, trehalose, glycerine, the VITAMIN.
---e drying:, obtain the solid state fermentation agent with the protectant bacteria suspension drying of interpolation.
Described drying step can be lyophilize:---the protectant bacteria suspension of interpolation is placed-60～-30 ℃ of refrigerators; after freezing 2.5～4 hours; freeze drier is put in taking-up; transfer condenser temperature-50～-40 ℃; slowly heat up through 5 ℃/30min; 3.5 take out the bacterium powder after～6 hours, obtain the solid state fermentation agent.
Also can be spraying drying:---the protectant bacteria suspension of interpolation is sent in the spray-dryer, 50～60 ℃ of inlet temperatures, 150～160 ℃ of temperature outs, flow velocity 0.1～0.3mL/s takes out the bacterium powder, obtains the solid state fermentation agent.
The application of a kind of probiotic lactobacillus starter in fresh cheese, it is characterized in that with in skimming milk, whey, lactose, the physiological saline one or more be activation solution, with the agent of exsiccant solid state fermentation with activation solution by activating, the activation solution consumption is that every gram is done agent 150～400mL, the activation back is as lactobacillus starter, adding to nonreactive sweet milk or reconstituted milk is during the routine of the cultured milk prods such as sour milk, cheese of raw material is produced, to make the cultured milk prod that contains profitable probliotics.
Can also add in the above-mentioned lactobacillus starter with lactobacterium casei starter volume ratio be one or more milk-acid bacterias in 2: 1～4 lactobacillus bulgaricus, thermophilus streptococcus, the dimethyl diketone suis, with lactobacterium casei together as lactobacillus starter, the interpolation total amount of starter accounts for 1%～5% of raw dairy, being applied to nonreactive sweet milk or reconstituted milk is during the routine of the cultured milk prods such as sour milk, cheese of raw material is produced, to make the cultured milk prod that contains profitable probliotics.
Advantage of the present invention and effect: the present invention is owing to adopted inherent probiotic bacterium in the human intestinal---and lactobacterium casei is as the starting strain of starter, and it is single and do not contain the situation of probiotic bacterium to have changed present conventional sour milk starter culture; And probiotic lactobacillus has been made highly active solid formulation, reduced starter in the cultured milk prod production process operation that spreads cultivation, saved fund, factory building, equipment and personnel's input, and made production operation be easier to control and management, the more stable quality of product.Present solid-state ferment agent for sour milk just contains conventional Yoghourt fermentation bacterial classification, and the solid lactic acid bacterium leavening agent made from probiotic bacterium of the present invention is with the accessibility used and the health perfect adaptation of product, cultured milk prod with this starter production, more help regulating the immunizing power of body, the health of promotion body, be higher than conventional ferment agent for sour milk aspect nutritive value and the nourishing function far away.Because the solid-state high reactivity probiotic lactobacillus agent in the production process of fresh cheese, adopted, significantly reduced in the fermenting process the management and the operation of bacterial classification, improved the nutritive value of production efficiency and product.
[embodiment]:
Embodiment 1:
The preparation method of probiotic lactobacillus starter, this method realizes by following steps:
---modified MRS (de Man, Rogosa, Sharpe Agar) culture medium preparation: with the deionized water is solvent, and volume ratio g/L adds following ingredients successively by weight: peptone 8, yeast extract paste 4, extractum carnis 2, glucose 6, sodium acetate 6, tween 80 1, ferric sulfate 1, manganous sulfate 2, triammonium citrate 2.
---insert lactobacterium casei: inoculum size 5%, 30 ℃ of culture temperature are added whey 5g/L in culturing process, and the pH value scope of control nutrient solution is 5.5～6.0, and incubation time 5～10 hours makes the bacterium number reach 10
9～10/ mL obtains the high-concentration culturing thing of lactobacterium casei.
---with centrifugal concentrate (the rotating speed 4000rpm/20min) of high-concentration culturing liquid.Abandoning supernatant.
---with the aqueous solution that 2.0% glycerine+12% skimming milk is made into as protective material, consumption be equivalent to centrifugal before high-concentration culturing liquid long-pending 1/5 (20%), be made into suspension with bacterial sediment thing uniform mixing.
---place-40 ℃ of refrigerator and cooled to freeze 3.5 hours the protectant bacteria suspension of interpolation after; freeze drier is put in taking-up, transfers condenser temperature-45 ℃, slowly heat up (5 ℃/30min); take out the bacterium powder after 5～5.5 hours, viable count can reach 700～1,200 hundred million/gram.This solid-state high reactivity probiotic lactobacillus bacterium powder can be used as the production that lactobacillus starter is used for milk-product, can also be applied in feed and the protective foods as immunologic active material.
Embodiment 2:
The prescription of modified MRS culture medium (g/L) is: peptone 10, yeast extract paste 3, extractum carnis 1, glucose 4, sodium acetate 2, tween 80 1, ferric sulfate 2, manganous sulfate 1, triammonium citrate 2.Insert lactobacterium casei, inoculum size 2%, 35 ℃ of culture temperature are added lactose 7g/L in culturing process, and the pH value scope of control nutrient solution is 5.0～5.5, incubation time 5～10 hours.With centrifugal concentrate (the rotating speed 5000rpm/10min) of high-concentration culturing liquid.Abandoning supernatant.Other parameter is with embodiment 1.
Embodiment 3:
Insert lactobacterium casei in the modified MRS culture medium in the foregoing description 1,2; inoculum size 3%; (feed supplement is whey or lactose to flow feeding; consumption is with embodiment 1 or 2); and control substratum acidity obtains high-concentration culturing liquid, and is centrifugal, with 10% sucrose-whey solution mixture as protective material; consumption be centrifugal before high-concentration culturing liquid long-pending 1/7 (14%), protective material and bacterial sediment thing uniform mixing are made into suspension.After placing-60 ℃ of refrigerator and cooled to freeze 2.5 hours the protectant bacteria suspension of interpolation, take out and put into freeze drier, transfer condenser temperature-50 ℃, slowly heat up (5 ℃/30min), take out the bacterium powder after 5.5～6 hours.Other parameter is with embodiment 1 or 2.
Embodiment 4:
Adopt initial medium; the growth temperature of regulating thalline is 37 ℃; flow feeding (with embodiment 3), and control substratum acidity obtains high-concentration culturing liquid, centrifugal; cultivate and add protective material; carry out spraying drying, 50～60 ℃ of inlet temperatures, 150～160 ℃ of temperature outs; flow velocity 0.1mL/s, the viable count in the bacterium powder are 200～60,000,000,000/gram.Other parameter is with embodiment 1,2 or 3.
Embodiment 5:
With the activation of 4% whey, the consumption 300mL/ gram of activator is done agent with the bacterium powder in embodiment 1,2,3 or 4, and viable count reaches 10～2,000,000,000/milliliter, as the milk-acid bacteria working stock culture.With the fresh skimmed milk of nonreactive is that the raw material making is the health fresh cheese of lactic-acid culture with the lactobacterium casei.Raw dairy is through 75 ℃/15s of pasteurize, add 0.015% calcium chloride, insert the lactobacterium casei starter 1～2% after activating, cultivate after 30 minutes for 31 ℃, add rennin, cut in the scope of curdled milk pH 4.40～4.85, leave standstill and begin after 15～20 minutes to stir and second-heating, temperature is brought up to 50 ℃～55 ℃ from 31 ℃～32 ℃ gradually; Discharge whey, add the cold water washing granular curd, make the temperature of granular curd reduce to 8 ℃～10 ℃, the washing triplicate.After leaching the moisture of granular curd, add 2%～4% salt, the container of packing into, sealing, 4 ℃ of refrigerations obtain containing the health fresh cheese of profitable probliotics.
Claims (9)
1, a kind of preparation method of probiotic lactobacillus starter is characterized in that this method realizes by following steps:
---a culture medium preparation: this substratum is solvent with the deionized water, and volume ratio g/L adds following ingredients successively by weight: peptone 8～10, yeast extract paste 3～5, extractum carnis 1～3, glucose 4～6, sodium acetate 2～6, tween 80 1～2, ferric sulfate 1～3, manganous sulfate 1～2, triammonium citrate 1～2;
---b spawn culture: with lactobacterium casei, add in the above-mentioned substratum by inoculum size 2～5%, culture temperature is 30～37 ℃, in the culturing process by weight volume ratio stream add carbon source 3-7g/L and/or nitrogenous source 3-5g/L, the pH value of control substratum is 5.0～6.5, incubation time 5～10 hours reaches 10 to the bacterium number
9～10/ mL obtains the high-concentration culturing thing of lactobacterium casei;
---c concentrates: above-mentioned high-concentration culturing thing is concentrated through rotating speed 3000～5000rpm/10～25min is centrifugal, obtain the enriched material of active lactobacillus casei;
---d adds protective material: in the enriched material of above-mentioned lactobacterium casei, add protective material, protectant consumption be in b step centrifugal before high-concentration culturing liquid long-pending 10～20%, make bacteria suspension;
---e drying:, obtain the solid state fermentation agent with the protectant bacteria suspension drying of interpolation.
2, the preparation method of probiotic lactobacillus starter according to claim 1, it is characterized in that also volume ratio g/L adds following material by weight: magnesium sulfate heptahydrate 1～3, dipotassium hydrogen phosphate 1～3 in the substratum preparation, Trisodium Citrate 2～5, four water manganous sulfates 2～4.
3, the preparation method of probiotic lactobacillus starter according to claim 1 and 2 is characterized in that:
---it is glucose, lactose, semi-lactosi, sucrose, maltose that described stream adds carbon source;
---it is whey, skimming milk, casein, soybean protein, amino acid that described stream adds nitrogenous source.
4, the preparation method of probiotic lactobacillus starter according to claim 3 is characterized in that:
---volume ratio stream is with Radix Dauci Sativae juice and/or each 10～20g/L of tomato juice, to promote the growth of thalline by weight.
5, the preparation method of probiotic lactobacillus starter according to claim 1 and 2 is characterized in that described protective material is one or more in sucrose, pectin, skimming milk, whey, serum albumin, trehalose, glycerine, the VITAMIN.
6, the preparation method of probiotic lactobacillus starter according to claim 1 and 2 is characterized in that described drying step can be lyophilize:
---the protectant bacteria suspension of interpolation is placed-60～-30 ℃ of refrigerators, after freezing 2.5～4 hours, take out and put into freeze drier; transfer condenser temperature-50～-40 ℃; slowly heat up through 5 ℃/30min, take out the bacterium powder after 3.5～6 hours, obtain the solid state fermentation agent.
7, the preparation method of probiotic lactobacillus starter according to claim 1 and 2 is characterized in that described drying step also can be a spraying drying:
---the protectant bacteria suspension of interpolation is sent in the spray-dryer, 50～60 ℃ of inlet temperatures, 150～160 ℃ of temperature outs, flow velocity 0.1～0.3mL/s takes out the bacterium powder, obtains the solid state fermentation agent.
8, the application of probiotic lactobacillus starter in fresh cheese of preparation in a kind of claim 1, it is characterized in that with in skimming milk, whey, lactose, the physiological saline one or more be activation solution, the agent of exsiccant solid state fermentation is activated with activation solution, the activation solution consumption is that every gram is done agent 150～400mL, activation back solution is as lactobacillus starter, adding to nonreactive sweet milk or reconstituted milk is during the routine of the cultured milk prods such as sour milk, cheese of raw material is produced, to make the cultured milk prod that contains profitable probliotics.
9, the application of probiotic lactobacillus starter according to claim 8 in fresh cheese, it is characterized in that adding with lactobacterium casei starter volume ratio be 2: one or more milk-acid bacterias in the lactobacillus bulgaricus of (1～4), thermophilus streptococcus, the dimethyl diketone suis, with lactobacterium casei together as lactobacillus starter, the interpolation total amount of starter accounts for 1%～5% of raw dairy, being applied to nonreactive sweet milk or reconstituted milk is during the routine of the cultured milk prods such as sour milk, cheese of raw material is produced, to make the cultured milk prod that contains profitable probliotics.
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| CN100360661C (en) * | 2005-11-30 | 2008-01-09 | 吉林大学 | Mutant strain of lactobacillus casei, its preparation method and fermentation production of L-lactic acid |
| CN100404665C (en) * | 2006-05-12 | 2008-07-23 | 河北农业大学 | Compound protective agent of lactic acid bacteria freeze-drying leavening agent |
| CN100415120C (en) * | 2006-01-18 | 2008-09-03 | 内蒙古蒙牛乳业（集团）股份有限公司 | Probiotics prepn., and its preparing method |
| CN100429980C (en) * | 2006-09-08 | 2008-11-05 | 肖雯娟 | Method for producing zymophyte agent in direct input type for yoghourt |
| CN101955900A (en) * | 2010-09-13 | 2011-01-26 | 北京金泰得生物科技股份有限公司 | Low-moisture solid culture and drying method for lactic acid bacteria |
| CN101971880A (en) * | 2010-11-30 | 2011-02-16 | 武汉光明乳品有限公司 | Fermented milk with high content of lactobacillus casei and manufacturing method thereof |
| CN102318683A (en) * | 2011-08-16 | 2012-01-18 | 吉林大学 | Fungal polysaccharide-containing probiotic cheese and preparation method thereof |
| CN101611135B (en) * | 2007-02-16 | 2012-06-20 | 雪印惠乳业株式会社 | Agent for improving viability of lactic acid bacterium |
| CN102907505A (en) * | 2011-08-02 | 2013-02-06 | 光明乳业股份有限公司 | Probiotics-containing soft fresh cheese and making method thereof |
| CN103141723A (en) * | 2013-03-13 | 2013-06-12 | 河北农业大学 | Preparation method of lactobacillus casei soybean yogurt direct vat set starter |
| CN103189499A (en) * | 2010-08-19 | 2013-07-03 | 株式会社明治 | Agent for improvement in survival of lactic acid bacterium and/or bifidobacterium |
| CN104031962A (en) * | 2014-05-05 | 2014-09-10 | 四川农业大学 | Optimized pEGF gene engineering Lactococcus lactis fermentation medium |
| CN106082436A (en) * | 2016-08-19 | 2016-11-09 | 赛富威环境科技南通有限公司 | Multi-dimensional composite carbon source composition and preparation method and application thereof |
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| CN107937304A (en) * | 2017-11-10 | 2018-04-20 | 北京颐和村生物科技有限公司 | The preparation method of the Yoghourt Production bacterium powder of lactic acid bacteria |
| CN108477381A (en) * | 2018-01-24 | 2018-09-04 | 福建傲农生物科技集团股份有限公司 | A kind of feeding beneficial bacteria of intestinal tract agent and its production method |
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| CN112980736A (en) * | 2021-03-31 | 2021-06-18 | 盐城维康生物科技有限公司 | Lactobacillus casei preparation capable of improving gastrointestinal function conditioning effect and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN100360661C (en) * | 2005-11-30 | 2008-01-09 | 吉林大学 | Mutant strain of lactobacillus casei, its preparation method and fermentation production of L-lactic acid |
| CN100415120C (en) * | 2006-01-18 | 2008-09-03 | 内蒙古蒙牛乳业（集团）股份有限公司 | Probiotics prepn., and its preparing method |
| CN100404665C (en) * | 2006-05-12 | 2008-07-23 | 河北农业大学 | Compound protective agent of lactic acid bacteria freeze-drying leavening agent |
| CN100429980C (en) * | 2006-09-08 | 2008-11-05 | 肖雯娟 | Method for producing zymophyte agent in direct input type for yoghourt |
| CN101611135B (en) * | 2007-02-16 | 2012-06-20 | 雪印惠乳业株式会社 | Agent for improving viability of lactic acid bacterium |
| CN103189499A (en) * | 2010-08-19 | 2013-07-03 | 株式会社明治 | Agent for improvement in survival of lactic acid bacterium and/or bifidobacterium |
| CN101955900A (en) * | 2010-09-13 | 2011-01-26 | 北京金泰得生物科技股份有限公司 | Low-moisture solid culture and drying method for lactic acid bacteria |
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| CN101971880A (en) * | 2010-11-30 | 2011-02-16 | 武汉光明乳品有限公司 | Fermented milk with high content of lactobacillus casei and manufacturing method thereof |
| CN101971880B (en) * | 2010-11-30 | 2012-07-25 | 武汉光明乳品有限公司 | Fermented milk with high content of lactobacillus casei and manufacturing method thereof |
| CN102907505A (en) * | 2011-08-02 | 2013-02-06 | 光明乳业股份有限公司 | Probiotics-containing soft fresh cheese and making method thereof |
| CN102318683A (en) * | 2011-08-16 | 2012-01-18 | 吉林大学 | Fungal polysaccharide-containing probiotic cheese and preparation method thereof |
| CN103141723A (en) * | 2013-03-13 | 2013-06-12 | 河北农业大学 | Preparation method of lactobacillus casei soybean yogurt direct vat set starter |
| CN103141723B (en) * | 2013-03-13 | 2014-01-22 | 河北农业大学 | Preparation method of lactobacillus casei soybean yogurt direct vat set starter |
| CN104031962A (en) * | 2014-05-05 | 2014-09-10 | 四川农业大学 | Optimized pEGF gene engineering Lactococcus lactis fermentation medium |
| CN106082436A (en) * | 2016-08-19 | 2016-11-09 | 赛富威环境科技南通有限公司 | Multi-dimensional composite carbon source composition and preparation method and application thereof |
| CN106754482A (en) * | 2016-11-28 | 2017-05-31 | 江苏梁丰食品集团有限公司 | A kind of preparation method of the probiotics leaven for producing functional sugar |
| CN107937304A (en) * | 2017-11-10 | 2018-04-20 | 北京颐和村生物科技有限公司 | The preparation method of the Yoghourt Production bacterium powder of lactic acid bacteria |
| CN108477381A (en) * | 2018-01-24 | 2018-09-04 | 福建傲农生物科技集团股份有限公司 | A kind of feeding beneficial bacteria of intestinal tract agent and its production method |
| CN108707568A (en) * | 2018-06-13 | 2018-10-26 | 河北康泰药业有限公司 | A kind of fermentation method for producing of mixing probiotics |
| CN112980736A (en) * | 2021-03-31 | 2021-06-18 | 盐城维康生物科技有限公司 | Lactobacillus casei preparation capable of improving gastrointestinal function conditioning effect and preparation method thereof |
| CN113632840A (en) * | 2021-08-12 | 2021-11-12 | 河北弗蒙特生物科技有限公司 | Probiotic milk or milk-containing beverage with high viable count and preparation method thereof |
| CN113881595A (en) * | 2021-10-09 | 2022-01-04 | 湖北工业大学 | Lactobacillus leaven containing protein fiber and preparation method thereof |
| CN113881595B (en) * | 2021-10-09 | 2023-08-25 | 湖北工业大学 | Lactic acid bacteria starter containing protein fiber and preparation method thereof |
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