CN101524146A - Method for improving number of probiotic live bacteria - Google Patents
Method for improving number of probiotic live bacteria Download PDFInfo
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- CN101524146A CN101524146A CN200910038553A CN200910038553A CN101524146A CN 101524146 A CN101524146 A CN 101524146A CN 200910038553 A CN200910038553 A CN 200910038553A CN 200910038553 A CN200910038553 A CN 200910038553A CN 101524146 A CN101524146 A CN 101524146A
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- lactic acid
- fermentation
- northern
- cordyceps
- live bacteria
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Abstract
The invention discloses a method for improving the number of probiotic live bacteria. North caterpillar fungus extracts are added into conventional lactic acid bacteria fermentation medium and the number of live bacteria is determined by adopting a method of conventional dilution plate anaerobic culture after fermentating and culturing lactic acid bacteria seeds for 8 to 24 hours. In the method, the north caterpillar fungus extracts obtained by hydrolysis and enzymolysis concentration are used as nutriments to be added into the conventional lactic acid bacteria fermentation medium, the north caterpillar fungus has rich nutrition, can provide rich nutrition for the growth of the probiotic and can obviously improve the number of live bacteria of the lactic acid bacteria in the fermentation broth.
Description
Technical field
The invention belongs to biotechnology, relate to a kind of method that improves number of live bacteria of probiotics specifically.
Background technology
Along with the raising of living standards of the people and the enhancing of health perception, people more and more pay close attention to diet to organism physiology or otherwise influence and sanatory effect, and the propensity to consume is just turning to the food with adequate nutrition and health care.Because probio has good regulating action to body enteron aisle fungus strain, enteron aisle fungus strain balance then is to be related to human nutrition and healthy vital factor, and this makes the research of probio become focus in the functional food research gradually.Lactic acid bacteria is an important physical probio in the human body intestinal canal, has effects such as keeping the normal microecological balance of intestines and stomach, the invasion that suppresses pathogen and infection and enhancing immunity of organisms, is the main little ecological class raw material of plurality kinds of health care food and medicine.Commercial product has: sour milk, lactic acid bacteria tablet, lactic acid bacteria powder etc.Mei Shi in 1882 finds lactic acid bacteria, and day in 1971, the Benson factory that suckles forever produced the bifid sour milk, and beginning is sold in the European market.French ST.Hubert company in 1986, the Benson factory that suckles forever introduces production technology from day, and bifidobacterium yoghurt has a large circulation freely.20th century the eighties, China begins to produce.
Oilgosaccharkdes is meant that oligosaccharides etc. is not digested and absorbs by the digestive ferment in the human body alimentary canal, but after entering human body intestinal canal, it can promote the growth of beneficial bacterium such as Bifidobacterium in the enteron aisle selectively, keep the general level of the health of human body, commercial product has: FOS, soyabean oligosaccharides, palatinose-oligosaccharides, oligoisomaltose etc.The Oilgosaccharkdes scope that is applied to the lactic acid bacteria product comprises: trehalose, FOS, oligoisomaltose, soyabean oligosaccharides, palatinose-oligosaccharides, xylo-oligosaccharide, oligomeric maltose.
The north Chinese caterpillar fungus also claims Cordyceps militaris, is the abbreviation of northern Chinese caterpillar Fungus, has another name called northern Chinese caterpillar Fungus, silkworm chrysalis Cordyceps sinensis etc.After the nineties in 20th century, many scholars have carried out big quantity research to domestication and the pharmacological action of northern Chinese caterpillar fungus, confirm that pharmacological action of northern Chinese caterpillar fungus and effect are identical with Cordyceps sinensis basically, can be used as the substitute of Cordyceps sinensis.Lung nourishing and qi invigorating arranged, increase smart kidney tonifying, hemostasis and phlegm, inhibition tumour, cough-relieving calmness, wind dispelling, heat radiation, stay Rong Meiyan, delay senility, tonifying kidney and strengthening yang, the reduction youth, effect and lung moistening and asthma relievings such as harmonious two sexual lifves, the enriching yin tonifying lung, enhance metabolism, improve the health, improve the effect of immunity of organisms.Studies show that, northern Chinese caterpillar Fungus contains chemical compositions such as cordycepin, cordycepic acid (sweet mellow wine), Cordyceps sinensis polysaccharide, protein, superoxide dismutase, amino acid, vitamin and various trace elements, and it of cordycepin has functions such as anticancer, antibiotic, antiviral; Cordycepic acid (sweet mellow wine) can fall low intracranial pressure, enhance metabolism thereby make cerebral hemorrhage and cerebral thrombosis disease to be eased significantly; Cordyceps sinensis polysaccharide is considered to one of extraordinary immunopotentiating agent on the our times.The north Chinese caterpillar fungus removes and contains cordycepin, D-sweet mellow wine, Cordyceps sinensis polysaccharide, adenosine, ergosterol, SOD enzyme, also contains protein more than 40%, and 18 seed amino acids, amino acid content are more than 25%, and nutrition is extremely abundant.
Early stage lactobacillus product still occurs with the liquid condition product, because viable count storage life weak point and number of viable in the liquid preparation are difficult to improve, makes the viable count in the product be difficult to guarantee.The all dead bacterium state of solid product occurs.Day the present invention's " biofermin " medicine treatment enterogastric diseases is obtained good result, but this medicine is dead bacteria preparation, after take " freeze-drying " technology production " new biofermin " again, but cost an arm and a leg.
Therefore, in the above-mentioned at home literature search, find no and utilize northern Cordyceps to promote lactobacillus-fermented, improve the bibliographical information of zymotic fluid viable count.
Summary of the invention
The purpose of this invention is to provide a kind of method that improves number of live bacteria of probiotics, because of probio bacterial classification nutritional requirement height, nourishing Northern cordyceps is abundant, and northern Cordyceps is added fermentation medium can provide abundant nutrition for the growth of probio, improves the viable count of probio in the zymotic fluid.
For achieving the above object, the present invention has taked following technical scheme:
A kind of method that improves number of live bacteria of probiotics may further comprise the steps:
1) preparation of northern Cordyceps
Add its quality 5-40 water doubly northwards in the Chinese caterpillar fungus dry product, smash pulping after boiling, at 70-98 ℃ of following hydrolysis 1-6 hour; The pH of hydrolyzate is adjusted into 7.5-10, adds the alkali protease of northern Chinese caterpillar fungus dry product quality 0.01-0.5%, at 40-75 ℃ of following enzymolysis 4-12 hour; Filtration, vacuum are concentrated into 5-20Brix, promptly get northern Cordyceps;
2) preparation of fermentation medium
In conventional fermentation lactic acid bacterium culture medium (MRS), by adding 0.1-5kg north Cordyceps in per 100 liters of conventional fermentation lactic acid bacterium culture mediums, accent pH is 5.0-7.5, sterilizes 15-40 minute for 100-121 ℃;
3) fermented and cultured of probio
When treating that culture medium is chilled to 10-35 ℃, with the inoculum concentration inoculation that three grades of seeds of lactic acid bacteria culturers are pressed culture medium cumulative volume 2-20%, single strain or close inoculation; The condition bottom fermentation that maintains temperature 10-45 ℃, mixing speed 5-100r/min, stuffiness or anaerobism, tank pressure 0.01-0.10MPa, pH 5.0-7.0 was cultivated 8-24 hour;
4) number of live bacteria of probiotics is measured
After fermented and cultured finishes, adopt conventional dilution plate methods,anaerobic to carry out viable count and measure.
Preferably, in the described step (1), the quality that adds alkali protease is the 0.02-0.2% of northern Chinese caterpillar fungus dry product quality.
Preferably, in the described step (2), by adding 0.5-2kg north Cordyceps in per 100 liters of conventional fermentation lactic acid bacterium culture mediums.
Lactic acid bacteria culturers is one or more in bifidobacterium bifidum, bifidobacterium infantis, bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, lactobacillus bulgaricus, lactobacillus acidophilus, Lactobacillus casei cheese subspecies, streptococcus thermophilus, the lactobacillus reuteri in the described step 3).
The present invention has following beneficial effect:
The present invention adds in the conventional lactobacillus-fermented culture medium as nutriment separate the concentrated northern Cordyceps that makes through Perhydrolase, because of probio bacterial classification nutritional requirement height, nourishing Northern cordyceps is abundant, northern Cordyceps is added fermentation medium can provide abundant nutrition for the growth of probio, can improve the viable count of Lactic Acid from Fermentation Broth bacterium significantly.
The specific embodiment
Below by specific embodiment the present invention is described.
Embodiment 1:
1. the preparation of northern Cordyceps
Get 2kg and do northern Chinese caterpillar fungus, add 40 premium on currency, pour beater into after boiling, pulled an oar 5 minutes, transfer pH to 9.5-10,95 ℃ of following hydrolysis 6 hours; The pH of hydrolysis slurry is adjusted into 8.5, adds the 4g alkali protease, 60 ℃ of following enzymolysis 5 hours; Filtration, vacuum are concentrated into 10Brix and promptly get northern Cordyceps;
2. probio bacterial classification
Lactobacillus acidophilus is carried out actication of culture to three grade seed, and culture medium is conventional lactic acid bacteria culture medium MRS, and cultural method is conventional lactic acid bacteria cultivation;
3. probiotics fermention is cultivated
Ferment respectively with two 50 liters of fermentation tanks (being numbered A1, A2), every canned fermentation medium 30-35 liter, A1 adds northern Cordyceps 1kg/100 liter in addition with conventional MRS culture medium, the A2 jar is done contrast with conventional lactic acid bacteria culture medium (MRS), respectively at 115 ℃ of sterilizations 20 minutes, be chilled to 35 ℃, three grades of seeds of lactobacillus acidophilus be inoculated into respectively by the inoculum concentration of fermentation medium cumulative volume 10% (v/v) carry out the stuffiness fermentation in two fermentation tanks, tank pressure is kept with filtrated air; Keep 37 ℃ of fermentation jar temperatures, pH5.5-6.5, mixing speed 30rpm/min, tank pressure 0.03MPa fermentation 18 hours;
4. the viable count in the mensuration zymotic fluid
Pour into methods,anaerobic from two jars of zymotic fluid samplings by flat board respectively and measure the bacterium number, A1 is 10
9-10
10Cfu/ml, A2 are 10
8-10
9Cfu/ml.
Embodiment 2:
1. the preparation of northern Cordyceps
Get 10kg and do northern Chinese caterpillar fungus, add 250 premium on currency, pour beater into after boiling, pulled an oar 5 minutes, transfer pH to 9.5-10,90 ℃ of following hydrolysis 6 hours; The pH of hydrolysis slurry is adjusted into 8.5, adds the alkali protease of 4g, 60 ℃ of following enzymolysis 5 hours; Filtration, vacuum are concentrated into 12Brix and promptly get northern Cordyceps;
2. probio bacterial classification
Lactobacillus bulgaricus is carried out actication of culture to three grade seed, and culture medium is conventional lactic acid bacteria culture medium, and cultural method is conventional lactic acid bacteria cultivation;
3. probiotics fermention is cultivated
Ferment respectively with two 50 liters of fermentation tanks (being numbered B1, B2), every canned fermentation medium 30-35 liter, B1 adds northern Cordyceps 4kg/100 in addition with conventional MRS culture medium and rises culture medium, the B2 jar is done contrast with conventional lactic acid bacteria culture medium (MRS), respectively at 115 ℃ of sterilizations 20 minutes, be chilled to 35 ℃, three grades of seeds of lactobacillus bulgaricus are inoculated into respectively by the inoculum concentration of fermentation medium cumulative volume 10% (v/v) carry out the stuffiness fermentation in two fermentation tanks, tank pressure is kept with filtrated air; Keep 37 ℃ of fermentation jar temperatures, pH5.5-6.5, mixing speed 30rpm/min, tank pressure 0.03MPa fermentation 18 hours;
4. the viable count in the mensuration zymotic fluid
Pour into methods,anaerobic from two jars of zymotic fluid samplings by flat board respectively and measure the bacterium number, B1 is 10
9-10
10Cfu/ml, B2 are 10
8-10
9Cfu/ml.
Embodiment 3:
1. the preparation of northern Cordyceps
Get 1kg and do northern Chinese caterpillar fungus, add 15 premium on currency, pour beater into after boiling, pulled an oar 5 minutes, transfer pH to 9.5-10,90 ℃ of following hydrolysis 6 hours; The pH of hydrolysis slurry is adjusted into 8.5, adds the alkali protease of 4g, 60 ℃ of following enzymolysis 5 hours; Filtration, vacuum are concentrated into 15Brix and promptly get northern Cordyceps;
2. probio bacterial classification
Streptococcus thermophilus, lactobacillus bulgaricus are carried out actication of culture to three grade seed respectively, and culture medium is conventional lactic acid bacteria culture medium, and cultural method is conventional lactic acid bacteria cultivation;
3. probiotics fermention is cultivated
(be numbered C1 with two 50 liters of fermentation tanks, C2) ferment respectively, every canned fermentation medium 30-35 liter, C1 adds northern Cordyceps 0.3kg/100 in addition with conventional MRS culture medium and rises culture medium, the C2 jar is done contrast with conventional lactic acid bacteria culture medium (MRS), respectively at 115 ℃ of sterilizations 20 minutes, be chilled to 35 ℃, with lactobacillus bulgaricus, 1: 1 by volume ratio of three grades of seeds of streptococcus thermophilus is mixed, mixed bacteria liquid is inoculated into respectively by the inoculum concentration of fermentation medium cumulative volume 10% (v/v) carries out the stuffiness fermentation in two fermentation tanks, tank pressure is kept with filtrated air; Keep 37 ℃ of fermentation jar temperatures, pH5.5-6.5, mixing speed 30rpm/min, tank pressure 0.03MPa fermentation 18 hours;
4. the viable count in the mensuration zymotic fluid
Pour into methods,anaerobic from two jars of zymotic fluid samplings by flat board respectively and measure the bacterium number, C1 is 10
9-10
10Cfu/ml, C2 are 10
8-10
9Cfu/ml.
The zymotic fluid number of live bacteria of probiotics that obtains according to the inventive method reaches 10
9More than the cfu/ml, and the viable count 10 in conventional MRS culture medium
8About cfu/ml.
Claims (4)
1. method that improves number of live bacteria of probiotics is characterized in that: may further comprise the steps:
1) preparation of northern Cordyceps
Add its quality 5-40 water doubly northwards in the Chinese caterpillar fungus dry product, smash pulping after boiling, 70-98 ℃ following hydrolysis 1-6 hour;
The pH of hydrolyzate is adjusted into 7.5-10, adds the alkali protease of northern Chinese caterpillar fungus dry product quality 0.01-0.5%, 40-75 ℃ following enzymolysis 4-12 hour; Filtration, vacuum are concentrated into 5-20Brix, promptly get northern Cordyceps;
2) preparation of fermentation medium
Add 0.1-5kg north Cordyceps in per 100 liters of conventional fermentation lactic acid bacterium culture mediums, accent pH is 5.0-7.5, and sterilization promptly gets fermentation medium;
3) fermented and cultured of probio
Treat that fermentation medium is cooled to 10-35 ℃, three grades of seeds of lactic acid bacteria culturers are pressed the inoculum concentration inoculation of fermentation medium cumulative volume 2-20%; The condition bottom fermentation that maintains 10-45 ℃, 5-100rpm/min, stuffiness or anaerobism, tank pressure 0.01-0.10MPa, pH5.0-7.0 was afterwards cultivated 8-24 hour.
2. the method for raising number of live bacteria of probiotics according to claim 1 is characterized in that: in the described step (1), the quality that adds alkali protease is the 0.02-0.2% of northern Chinese caterpillar fungus dry product quality.
3. the method for raising number of live bacteria of probiotics according to claim 1 is characterized in that: in the described step (2), add 0.5-2kg north Cordyceps in per 100 liters of conventional fermentation lactic acid bacterium culture mediums.
4. the method for raising number of live bacteria of probiotics according to claim 1, it is characterized in that: in the described step (3), lactic acid bacteria culturers is one or more in bifidobacterium bifidum, bifidobacterium infantis, bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, lactobacillus bulgaricus, lactobacillus acidophilus, Lactobacillus casei cheese subspecies, streptococcus thermophilus, the lactobacillus reuteri.
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| CN200910038553A CN101524146A (en) | 2009-04-10 | 2009-04-10 | Method for improving number of probiotic live bacteria |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102266054A (en) * | 2011-07-26 | 2011-12-07 | 西藏月王生物技术有限公司 | Aweto ferment and preparation method thereof |
| CN101731511B (en) * | 2010-02-03 | 2013-04-03 | 徐赟姣 | Probiotic active product and preparation method thereof |
| CN106036856A (en) * | 2016-06-15 | 2016-10-26 | 杭州彧弋科技有限公司 | High-solubility probiotics fermentation dendrobium nobile powder and preparation and application thereof |
| CN107047764A (en) * | 2017-01-12 | 2017-08-18 | 江苏君乐宝乳业有限公司 | A kind of Cordyceps militaris enzyme yogurt and preparation method thereof |
| CN109609430A (en) * | 2018-12-07 | 2019-04-12 | 吉林省浦生泰生物技术有限责任公司 | A kind of grain culture medium promoting growth of probiotics based on Chinese medicine additive |
| CN109876092A (en) * | 2019-01-24 | 2019-06-14 | 金活医药集团有限公司 | Chinese medicine prebiotic compositions, the culture medium of the prebiotics containing Chinese medicine, Chinese medicine prebiotics and product of probiotic combinations and preparation method thereof |
| CN113373079A (en) * | 2021-03-31 | 2021-09-10 | 河北峰宇生物技术有限责任公司 | Method for increasing viable count and prolonging storage time of lactobacillus liquid |
| CN114052219A (en) * | 2020-08-06 | 2022-02-18 | 重庆康康康生物科技有限公司 | Preparation method and application of leech fermentation liquor |
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2009
- 2009-04-10 CN CN200910038553A patent/CN101524146A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101731511B (en) * | 2010-02-03 | 2013-04-03 | 徐赟姣 | Probiotic active product and preparation method thereof |
| CN102266054A (en) * | 2011-07-26 | 2011-12-07 | 西藏月王生物技术有限公司 | Aweto ferment and preparation method thereof |
| CN102266054B (en) * | 2011-07-26 | 2013-01-30 | 西藏月王生物技术有限公司 | Fermentation functionality aweto beverage and preparation method thereof |
| CN106036856A (en) * | 2016-06-15 | 2016-10-26 | 杭州彧弋科技有限公司 | High-solubility probiotics fermentation dendrobium nobile powder and preparation and application thereof |
| CN106036856B (en) * | 2016-06-15 | 2019-06-04 | 杭州彧弋科技有限公司 | A kind of highly dissoluble probiotics fermention dendrobe powder and its preparation and application |
| CN107047764A (en) * | 2017-01-12 | 2017-08-18 | 江苏君乐宝乳业有限公司 | A kind of Cordyceps militaris enzyme yogurt and preparation method thereof |
| CN109609430A (en) * | 2018-12-07 | 2019-04-12 | 吉林省浦生泰生物技术有限责任公司 | A kind of grain culture medium promoting growth of probiotics based on Chinese medicine additive |
| CN109876092A (en) * | 2019-01-24 | 2019-06-14 | 金活医药集团有限公司 | Chinese medicine prebiotic compositions, the culture medium of the prebiotics containing Chinese medicine, Chinese medicine prebiotics and product of probiotic combinations and preparation method thereof |
| CN114052219A (en) * | 2020-08-06 | 2022-02-18 | 重庆康康康生物科技有限公司 | Preparation method and application of leech fermentation liquor |
| CN113373079A (en) * | 2021-03-31 | 2021-09-10 | 河北峰宇生物技术有限责任公司 | Method for increasing viable count and prolonging storage time of lactobacillus liquid |
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Effective date of registration: 20101210 Address after: 510095 No. 100 martyrs Middle Road, Guangdong, Guangzhou Applicant after: Guangdong Institute of Microbiology Co-applicant after: Dongguan Research Center of Agricultural Science Address before: 510095 No. 100 martyrs Middle Road, Guangdong, Guangzhou Applicant before: Guangdong Institute of Microbiology Co-applicant before: Huankai Microbes Tech Co., Ltd., Guangdong |
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Application publication date: 20090909 |