CN1315417C - Kluyveromyces marxianus yeast fermented drink and its preparation method - Google Patents
Kluyveromyces marxianus yeast fermented drink and its preparation method Download PDFInfo
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Abstract
The present invention relates to a Kluyveromyces marnianu fermenting drink and a preparation method thereof, which is characterized in that a strain of separated Kluyveromyces marnianu microzyme is inoculated in screened culture media at the temperature of 25 to 30 DEG C and then processed through fermentation cultivation, aseptic filtration and subsequent conventional treatment so as to obtain the drink. The preparation method comprises the following steps: (1) separating and cultivating strains; (2) preparing strain fermenting solutions; (3) preparing primary fermentation solutions; (4) preparing original fermentation solutions; (5) filtering; (6) testing effective components; (7) seasoning with white granulated sugar so as to obtain a sweet sour and delectable flavour; (8)filling; (9)sterilizing. The present invention provides the Kluyveromyces marnianu fermenting drink which is a healthy drink with natural activities and the preparation method thereof through research and development with the advantages of solved problem of single nutrition and excessive manual component adjunction existing in beverage industries at present, integrally integrated nutrition property and health care property, natural component without any manual additive and no toxic side effect on organisms.
Description
Affiliated field
The present invention relates to the fermentation industry ambit, is a kind of Marx's Crewe dimension fermented beverage and preparation method thereof specifically
Background technology
As everyone knows, in food industry, the product of many fungi fermentations can be used as many country and peoples' cuisines or distinctive flavouring.Different bacterial classifications is given different tunnings, is indebted to the mankind with different nutritive values.Kluyveromyces marxianus Kluyveromyces marnianus (E.G.Hansen) van der Walt is a kind of useful yeast fungus of the mankind both domestic and external that is popular in, and is widely used in the production of yogurt making and some fruit wine drink.Research and related data according to us show: with its cultivation and fermentation in the good environment of artificial design, can produce and secrete trophic factors and the bioactivator of multiple beneficial in health.We utilize its this characteristic, are developed into the healthy natural beverage of a kind of novel green, believe that it to safeguarding health and improving people's quality of life, plays a positive role.
Documents and materials proved already: in the cell of yeast class fungi, contain following multiple important nutrition and active material.(1) Yeast protein, its catabolite are yeast nuclein, and it is the important nutrient of living organism, are again to be used for the treatment of the medicine that trembles and benumb clinically; (2) yeast nucleic acid, in yeast cells, it exists with the prothetic group form of Yeast protein, contains phosphoric acid, purine and pyrimidine, effects such as having the expansion peripheral vessel clinically, improve ferroheme concentration and red blood cell number, alleviate edema, be antiviral.In addition, studies have shown that it also has anticancer and function dissolved cancer cell; (3) glucocinin, be otherwise known as yeast insulin or glucokinin, its same blood insulin, globulin insulin, ox pancreas insulin performance are similar, can effectively correct insulin deficit disease---the dysbolism of diabetes; (4) rich in amino acid in Yeast protein, has eight necessary seed amino acids of human body.Especially the content of lysine surpasses any other protein.(5) multivitamin in yeast cells and metabolite thereof, contains a large amount of B family vitamins and vitamin A, D, E, C.(6) bioactivator, foremost have two kinds, first yeast ferment, another is a beta glucan.Yeast ferment exists with yeast nucleic acid ferment, interior tryptose ferment, interior neutral protein ferment various ways.They both can promote the food hydrolysis, were converted into the digestive pharmaceutical that is of value to human nutrition, were again the drug ingedients of control cholecystitis, pancreatitis and other intestines problems; Beta glucan is that yeast is secreted a kind of biochemical substances that produces in its metabolic process.According to the literature, it is a kind of cancer-resisting substance universally acknowledged at present and that extremely pay attention to; (7) lot of organic acids can be aid digestion, corrosion-proof and sterilization, and killing pathogenic bacteria has the effect of good cleaning intestines and stomach.
According to of the research of present Chinese scholars to Marx's Crewe dimension bacterium.Verified, its producible natural component has: bioactivators such as nucleotides, compound sugar, oligopeptides, agglutinin, antibiotic and plurality of enzymes.Nucleotides is the fundamental structural unit of nucleic acid, is material very important in the metabolism, almost participates in all biochemical reactions of cell.Studies have shown that in a large number nucleic acid can be regulated metabolism, improves immunity of organisms, improves liver function, reduces physical consumption, improves functions of intestines and stomach, improves sleep, antifatigue, anti-oxidant and bringing into play important effect at aspect such as promote to grow; Compound sugar can not only reduce the absorption of cholesterol, can also promote the growth of probios such as Bifidobacterium Bifidum, lactic acid bacteria in the intestines, thereby reaches the effect of prevention enteritis.In addition, all metabolism has important facilitation to human-body biological for oligopeptides, agglutinin, antibiotic and various enzyme.
Make a general survey of present beverage market, no matter all kinds of Juices that liked by the consumer, or carbonated beverage, and the fruit vinegar beverage that is seen in report, they or nutrition are single, or its composition all is to add the synthetic material, all can not satisfy the modal need that current people's nutrition and health care combines.
Summary of the invention
The purpose of this invention is to provide a kind of overcome the single and artificial adding ingredient of nutrition in the present beverage industry cross indiscriminate problem and research and develop a kind ofly integrate nutrition, keep healthy, and its composition does not contain any artificial additive matter from natural, to organism natural organic active drink without any side effects-Marx's Crewe dimension fermented beverage and preparation method thereof.
A kind of Marx's Crewe dimension fermented beverage, it is characterized in that adopting strain kluyveromyces marxianus bacterium--a Kluyveromyces marxianus, under aseptic condition, be inoculated in the fermentation broth, raw materials used and the proportioning of described culture medium is: potato soup 50-70 part, 5 parts of glucose, maltose 0.1-0.5 part, 0.1 part of lactose, starch sugar 2-5 part, carrot juice 3-5 part also add water to 1000 parts, standing for fermentation is three days under 25-30 ℃ of condition, makes zymotic fluid just; Just zymotic fluid is being lower than under 20 ℃ of conditions, and standing for fermentation three days is made through aseptic filtration and follow-up conventional treatment then.
Preparation method and process are as follows:
The separation of A bacterial classification is cultivated
Under aseptic, room temperature condition, will be from " hiding clever mushroom " bacterium among the people through sterilization PBS washing three times, collude with platinum ear circle and to get its deep colony, be inoculated on the isolation medium, put then in 37 ℃ of constant incubators, through cultivation in 5-7 days; Select the perusal bacterium colony to be the cheese shape, milky or dark brown, surface smoothing, glossy, the edge is smooth, and microscopic examination is a large amount of mycelia and the independent bacterium colony of spore.Do secondary separation under the aseptic condition, used culture medium is the same, cultivates 3-5 days in rearmounted 37 ℃ of constant incubators, selects the same independent bacterium colony, is inoculated under the aseptic condition and preserves on the culture medium preservation under 4 ℃ of conditions;
Above-mentioned isolation medium is by following raw material and proportioning preparation:
10 parts of 200 parts of glucose 10-15 of potato soup part peptones
2 parts of agar of yeast extract add water to 1000 parts for 20 parts
Above-mentioned preservation culture medium is by following raw material and proportioning preparation:
10 parts of 200 parts of glucose 5-7 of potato soup part peptones
Agar adds water to 1000 parts for 20 parts
The preparation of B fermented bacterium liquid
The fermented bacterium liquid culture medium uses following raw material and proportioning preparation:
Potato soup 200-300 part glucose 15-20 part peptone 10-20 part
Maltose 2-10 part adds water to 1000 parts
1. I level fermented bacterium liquid: will be stored in 4 ℃ of kluyveromyces marxianus bacterial classifications under the condition, be inoculated under the aseptic condition in the fermented bacterium liquid culture medium test tube, cultivate 2-3 days in the 25-30 ℃ of constant incubator, I level fermented bacterium liquid;
2. II level fermented bacterium liquid: with I level fermented bacterium liquid, be inoculated under the aseptic condition in the bottled fermented bacterium liquid culture medium of 250ML, under 25-30 ℃ of constant temperature, cultivated 2-3 days;
C is preparation of fermentation liquid just:
With II level strain liquid, be inoculated in the fermentation culture under the aseptic condition, standing for fermentation is three days under the 25-30 ℃ of condition; Raw materials used and the proportioning of its culture medium is:
Potato soup 50-70% glucose 5% maltose 0.1-0.5% lactose 0.1%
Starch sugar 2-5% carrot juice 3-5% adds water to 1000
The preparation of D original fermentation liquor:
The first zymotic fluid that C is made under 20 ℃ of following conditions, standing for fermentation three days, drink stoste;
E filters;
The F seasoning;
The G can;
The H sterilization.
This drink has been concentrated the multiple advantage of current market popularity various types of beverages, and its making, composition, color and luster, taste all have own unique style.Following several respects are generally arranged: (1) trophism, this drink contains the nutritional labeling of multiple needed by human body, as protein, fat, sugar, amino acid, compound sugar, vitamin A, D, E, B, C and various trace element, mineral matter, organic acid (seeing the attached inspection report in back for details), can replenish human nutrition needs comprehensively; (2) health as previously mentioned, in the saccharomycete growth course, can produce the active factors (because present Food Inspection content and method limit, some do not listed inspection content) of multiple beneficial in health.In addition, the zoopery interpretation of result according to us proves that this drink has certain antagonism fat deposit and reduces the effect of blood sugar; (3) natural sex, the active material of a large amount of useful human bodies that this drink contains is as nucleotides, compound sugar, oligopeptides, various enzyme, various vitamin, mineral matter, organic acid etc.They or kluyveromyces marxianus bacterium produce in its growth metabolism process and secrete, or raw material natural component institute is intrinsic, is not to be derived from artificial interpolation.They need and keep health to replenishing human nutrition, and are significant.In addition, we compare with other drinks of current market popularity by the novel drink of development, and are not only nutritious, are sheerly naturally, and taste pure just, easily accepted by the consumer.By our repeated experiments repeatedly successively, prove that its stable components, taste are constant, good reproducibility and animal subject had no side effect, can be used in suitability for industrialized production.Along with China builds the propelling of moderately well-off paces and the raising of social people's health perception, this drink will be subjected to social people's bigger favor, on economy, technology and society effect, all will produce positive effect.
Description of drawings
Fig. 1 is the influence of drink Normal juice of the present invention to normal rat hemoglobin
Fig. 2 is the influence of drink Normal juice to normal rat red blood cell sum
Fig. 3 is the influence of drink Normal juice to normal rat leukocyte sum
The experimental result explanation
The research of 1 fat reducing effect---prevention obese model method
Kunming mouse is divided into blank group (common drinking-water group), model control group (culture medium juice at random Group), reach three given the test agent experimental group (drink Normal juice group, HIGH PRESSURE TREATMENT group, high-pressure filteration group). From reality Test beginning, culture medium juice group, drink Normal juice group, HIGH PRESSURE TREATMENT group, the every treated animal of high-pressure filteration group give etc. The beverage of amount, common drinking-water group gives general running water in the same manner. Raised 30 days continuously, cut open the belly and get body Fat (testis and perirenal fat pad) is also weighed, and calculates fat body ratio. Fat body ratio=fat weight/body weight * 100. Relatively three groups of fat reducing effects take common drinking-water group, culture medium juice group as contrast, are carried out t to each experimental group number certificate Check.
Table one: fat reducing effect
| Group | Dosage (ml/ group) | Number of animals (individual) | Body fat value (g) |
| Common drinking-water group | 25 | 5 | 1.58±0.34 |
| Culture medium juice group drink Normal juice group HIGH PRESSURE TREATMENT group high-pressure filteration group | 25 25 25 25 | 5 5 5 5 | 1.66±0.54 0.44±0.16** # # 0.55±0.09** # # 0.89±0.28* # |
Annotate: compare *: P＜0.05, * *: P＜0.01 with common drinking-water group; Compare with culture medium juice group,#：
P＜0.05，
##：P＜0.01
Table two: fat body ratio
| Group | Dosage (ml/ group) | Number of animals (individual) | The fat body is than (%) |
| Common drinking-water group culture medium juice group drink Normal juice group HIGH PRESSURE TREATMENT group high-pressure filteration group | 25 25 25 25 25 | 5 5 5 5 5 | 4.2±0.7 5.0±1.5 1.4±0.6** ## 1.7±0.2** ## 2.8±0.8* # |
Annotate: compare *: P＜0.05, * *: P＜0.01 with common drinking-water group; Compare with culture medium juice group,#：
P＜0.05，
##：P＜0.01
By table one, table two as can be known, in the experiment of prevention obese model, as the common drinking-water group of blank, body Fat is 1.58g, and fat/body is 4.2%, and as the culture medium juice group of model contrast, body fat is 1.66g, fat/ Body is that 5.0%, two control group does not have significant difference. Under the similarity condition, drink Normal juice group, body fat is 0.44g, Fat/body is 1.4%; HIGH PRESSURE TREATMENT group, body fat are 0.55g, and fat/body is 1.7%; The high-pressure filteration group, body fat Be 0.89g, fat/body is 2.8%, compares with two control groups, and certain fat reducing effect is arranged. Through the t check analysis, Drink Normal juice group, HIGH PRESSURE TREATMENT group are compared with two control groups, and difference all reaches utmost point significant difference level, high pressure mistake The filter group reaches the significance level of difference.
The test of 2 hypoglycemics---reduce the fasting blood-glucose experiment
After the Kunming mouse fasting 24 hours, give alloxan (150mg/Kg) moulding, prohibit after 5-7 days Ate 3-5 hour, and detected its blood glucose value 10-25mmol/L person and be hyperglycemia model success animal. Select hyperglycaemia Animal pattern divides into groups by fasting 3-5 hour blood sugar level, is divided at random 4 groups, all conventional raisings, nature Illumination. (1) common drinking-water group is given running water 100ml/ group every day; (2) 10 times of drinks concentrate group and give every day With 10 times of drink concentrate 100ml/ groups; (3) drink Normal juice group is given drink Normal juice 100ml/ group every day; (4) 50% dilution Normal juice group is given 50% dilution Normal juice 100ml/ group every day. Experiment is 30 days continuously, and blood is got in docking, Survey fasting blood sugar (fasting is front with experiment), relatively each treated animal blood glucose value and blood sugar decline percentage. Blood sugar Blood glucose value * 100% before decline percentage=(blood glucose value after blood glucose value before the experiment-experiment)/experiment. Compare the three Blood sugar decreasing effect take common drinking-water group as contrast, is checked according to carrying out t each experimental group number.
Table three: blood sugar decreasing effect
| Group | Dosage (ml/ group) | Number of animals (individual) | Hypoglycemic value (mmol/L) | Blood sugar rate of descent (%) |
| 10 times of concentrated group Normal juice group 50% dilution groups of common drinking-water group | 50 50 50 50 | 8 8 9 8 | 1.13±0.12 3.96±0.79** 3.40±0.68** 2.93±0.62* | 15.44±0.46 42.15±9.51** 35.05±9.12** 27.95±9.52* |
Annotate: compare *: P＜0.05, * *: P＜0.01 with common drinking-water group
As shown in Table 3, the diabetes model of successfully inducing, the average blood sugar value reaches 12.45mmol/L. Common The drinking-water group in contrast, its blood glucose value 1.13mmol/L that descended, the blood sugar rate of descent is 15.44%. Same batten Under the part, 10 times concentrate group, the blood glucose value 3.96mmol/L that descended, and the blood sugar rate of descent is 42.15%; The Normal juice group, The blood glucose value 3.40mmol/L that descended, the blood sugar rate of descent is 35.05%; 50% dilution group, blood glucose value has descended 2.93mmol/L the blood sugar rate of descent is 27.95%. Through the t check analysis, all experiment beverage group and control groups Compare, difference all reaches the level of signifiance, illustrates that drink has certain blood sugar reducing function. By hypoglycemic effect, three Have certain dosage effect between the concentration, concentration is more high, and hypoglycemic effect is more good.
3 hide clever mushroom drink to the impact of HC, red blood cell sum, total white blood cells in the rat blood
Give drink Normal juice with the Wister rat after conventional raising a period of time, tail vein blood is being given Give HC, red blood cell sum, the total white blood cells surveyed respectively before and after the drink in its blood. Relatively The content of indices in the blood is to give before the drink carrying out the t check as contrast.
Table four: hemoglobin, red blood cell, leukocytic impact
| Time | Hemoglobin | Red blood cell | Leucocyte |
| (g/100ml) | (10 6/mm 3) | (10 3/mm 3) | |
| After giving to give drink before the drink | 13.86±0.96 13.96±0.29 | 8.53±0.78 8.27±0.68 | 14.86±0.47 14.75±0.75 |
By table four know give drink Normal juice after, the HC in the rat blood, red blood cell sum, white TCS does not have marked change, illustrate give drink Normal juice after, rat has kept normal physiological status, Illustrate also that from a side this drink is as the security of beverage.
The mensuration of 4 hemoglobins---acidization
The Wister rat is divided into blank group and sample experimental group at random, give respectively common drinking-water and Drink Normal juice also gives the conventional feed raising. Per seven days tail vein bloods are once given the blood acid adding, make blood red Albumen becomes brown acid haematin, and dilution normal glass look post rear and chromocytometer carries out order The light ratio look namely gets the grams of contained hemoglobin in every 100ml blood. Compare the two content of hemoglobin, To take common drinking-water group as contrast, experimental group number is checked according to carrying out t.
Known by figure one, give drink after, the content of hemoglobin in the rat blood is not compared not aobvious with control group Work property difference, thus the explanation drink is to the not impact of its content of hemoglobin.
5 erythrocytic mensuration
The Wister rat is divided into blank group and sample experimental group at random, give respectively common drinking-water and Drink Normal juice also gives the conventional feed raising. Per seven days tail vein bloods once carry out red blood cell count(RBC), namely Get every cubic millimeter of (mm3) contained red blood cell sum in the blood. Compare the two red blood cell total quantity, with general Logical drinking-water group is contrast, carries out the t check.
Known by figure two, give drink after, the red blood cell sum in the rat blood is not compared with control group not significantly The property difference, thereby the explanation drink is to the not impact of its red blood cell sum.
The mensuration of 6 total white blood cellses
The Wister rat is divided into blank group and sample experimental group at random, gives common drinking-water and drink Normal juice respectively and give conventional feed to raise.Per seven days tail vein bloods once, acidification is carried out white blood cell count(WBC), promptly gets every cubic millimeter of (mm
3) contained total white blood cells in the blood.Relatively the two total white blood cells is contrast with common drinking-water group, and the test group data are carried out the t check.
Know by figure three, give drink after, the total white blood cells in the rat blood is compared with control group does not have significant difference, thus the explanation drink is to the not influence of its total white blood cells.
The specific embodiment
Below in conjunction with embodiment the present invention is described in further details:
Embodiment 1:
On October 9th, 2002 was 021009 drink 50000ml according to the conventional ratio preparation lot number of each component of culture medium,
(1) separation of bacterial classification is cultivated
Under aseptic, room temperature condition, will be from " hiding clever mushroom " bacterium among the people through sterilization PBS washing three times, collude with platinum ear circle and to get its deep colony, be inoculated on the isolation medium, put then in 37 ℃ of constant incubators, through cultivation in 5 days; Select the perusal bacterium colony to be the cheese shape, milky or dark brown, surface smoothing, glossy, the edge is smooth, and microscopic examination is a large amount of mycelia and the independent bacterium colony of spore.Do secondary separation under the aseptic condition, used culture medium is the same, cultivates 3 days in rearmounted 37 ℃ of constant incubators, selects the same independent bacterium colony, is inoculated under the aseptic condition and preserves on the culture medium preservation under 4 ℃ of conditions;
The raw material of isolation medium and proportioning are:
Potato soup 200ml glucose 10g peptone 10g yeast extract 2g agar 20g adds water to 1000ml
The raw material and the proportioning of preserving culture medium are:
Potato soup 200ml glucose 5g peptone 10g agar 20g adds water to 1000ml
2) preparation of fermented bacterium liquid
The fermented bacterium liquid culture medium uses following raw material and proportioning preparation:
Potato soup 200ml glucose 15g peptone 20g maltose 10g adds water to 1000ml
1. I level fermented bacterium liquid: will be stored in 4 ℃ of kluyveromyces marxianus bacterial classifications under the condition, be inoculated under the aseptic condition in the fermented bacterium liquid culture medium test tube, cultivate 3 days in 25 ℃ of constant incubators, I level fermented bacterium liquid;
2. II level fermented bacterium liquid: with I level fermented bacterium liquid, be inoculated under the aseptic condition in the bottled fermented bacterium liquid culture medium of 250ML, under 25 ℃ of constant temperatures, cultivated 3 days;
(3) first preparation of fermentation liquid:
With II level strain liquid, be inoculated under the aseptic condition in the zymotic fluid culture medium, standing for fermentation is 3 days under 25 ℃ of conditions; Raw materials used and the proportioning of its culture medium is:
Potato soup 25000ml
Glucose 2500g
Maltose 250g
Lactose 50g
Starch sugar 250g
Carrot juice 1500ml
Water 23500ml
(4) original fermentation liquor: the first zymotic fluid that (3) are made is being lower than under 20 ℃ of conditions, and standing for fermentation 3 days gets drink stoste;
(5) filter;
(6) active ingredient check;
(7) seasoning transfers to sour and sweet palatability with white granulated sugar;
(8) can;
(9) sterilization.
Embodiment 2:
On March 14th, 2003 was 030314 drink 100000ml according to the optimal proportion of each component preparation lot number, and its proportioning is:
(1) separation of bacterial classification is cultivated
Under aseptic, room temperature condition, will be from " hiding clever mushroom " bacterium among the people through sterilization PBS washing three times, collude with platinum ear circle and to get its deep colony, be inoculated on the isolation medium, put then in 37 ℃ of constant incubators, through cultivation in 7 days; Select the perusal bacterium colony to be the cheese shape, milky or dark brown, surface smoothing, glossy, the edge is smooth, and microscopic examination is a large amount of mycelia and the independent bacterium colony of spore.Do secondary separation under the aseptic condition, used culture medium is the same, cultivates 5 days in rearmounted 37 ℃ of constant incubators, selects the same independent bacterium colony, is inoculated under the aseptic condition and preserves on the culture medium preservation under 4 ℃ of conditions;
The raw material of isolation medium and proportioning are:
Potato soup 200ml glucose 15g peptone 10g yeast extract 2g agar 20g adds water to 1000ml
The raw material and the proportioning of preserving culture medium are:
Potato soup 200ml glucose 7g peptone 10g agar 20g adds water to 1000ml
2) preparation of fermented bacterium liquid
The fermented bacterium liquid culture medium uses following raw material and proportioning preparation:
Potato soup 300ml glucose 20g peptone 10g maltose 2g adds water to 1000ml
1. I level fermented bacterium liquid: will be stored in 4 ℃ of kluyveromyces marxianus bacterial classifications under the condition, be inoculated under the aseptic condition in the fermented bacterium liquid culture medium test tube, cultivate 2 days in 30 ℃ of constant incubators, I level fermented bacterium liquid;
2. II level fermented bacterium liquid:, in the aseptic condition fermented bacterium liquid culture medium that the sub-250ML of inoculation is bottled down, under 30 ℃ of constant temperatures, cultivated 2 days with I level fermented bacterium liquid;
(3) first preparation of fermentation liquid:
With II level strain liquid, be inoculated under the aseptic condition in the zymotic fluid culture medium, standing for fermentation is 3 days under 30 ℃ of conditions; Raw materials used and the proportioning of its culture medium is:
Potato soup 70000ml
Glucose 7000g
Maltose 300g
Lactose 100g
Starch sugar 500g
Carrot juice 5000ml
Water 25000ml
(4) original fermentation liquor: the first zymotic fluid that (3) are made is being lower than under 20 ℃ of conditions, and standing for fermentation three days gets drink stoste;
(5) filter;
(6) active ingredient check;
(7) seasoning transfers to sour and sweet palatability with white granulated sugar;
(8) can;
(9) sterilization.
Embodiment 3:
On June 10th, 2003 was 030610 drink 200000ml according to the optimized proportion of each component of culture medium preparation lot number, and its proportioning is:
(1) separation of bacterial classification is cultivated
Under aseptic, room temperature condition, will be from " hiding clever mushroom " bacterium among the people through sterilization PBS washed twice, collude with platinum ear circle and to get its deep colony, be inoculated on the isolation medium, put then in 37 ℃ of constant incubators, through cultivation in 5 days; Select the perusal bacterium colony to be the cheese shape, milky or dark brown, surface smoothing, glossy, the edge is smooth, and microscopic examination is a large amount of mycelia and the independent bacterium colony of spore.Do secondary separation under the aseptic condition, used culture medium is the same, cultivates 3 days in rearmounted 37 ℃ of constant incubators, selects the same independent bacterium colony, is inoculated under the aseptic condition and preserves on the culture medium preservation under 4 ℃ of conditions;
The raw material of isolation medium and proportioning are:
Potato soup 200ml glucose 10g peptone 10g yeast extract 2g agar 20g adds water to 1000ml
The raw material and the proportioning of preserving culture medium are:
Potato soup 200ml glucose 5g peptone 10g agar 20g adds water to 1000ml
2) preparation of fermented bacterium liquid
The fermented bacterium liquid culture medium uses following raw material and proportioning preparation:
Potato soup 200ml glucose 15g peptone 20g maltose 10g adds water to 1000ml
1. I level fermented bacterium liquid: will be stored in 4 ℃ of kluyveromyces marxianus bacterial classifications under the condition, be inoculated under the aseptic condition in the fermented bacterium nutrient solution test tube, cultivate 3 days in 25 ℃ of constant incubators, I level fermented bacterium liquid;
2. II level fermented bacterium liquid: with I level fermented bacterium liquid, be inoculated under the aseptic condition in the bottled fermented bacterium nutrient solution of 250ML, under 25 ℃ of constant temperatures, cultivated 3 days;
(3) first preparation of fermentation liquid:
With II level strain liquid, be inoculated in the fermentation culture under the aseptic condition, standing for fermentation is 3 days under 25 ℃ of conditions; Raw materials used and the proportioning of its culture medium is:
Potato soup 25000ml
Glucose 2500g
Maltose 250g
Lactose 50g
Starch sugar 250g
Carrot juice 1500ml
Water 23500ml
(4) original fermentation liquor: the first zymotic fluid that (3) are made is being lower than under 20 ℃ of conditions, and standing for fermentation 3 days gets drink stoste;
(5) filter;
(6) active ingredient check;
(7) seasoning transfers to sour and sweet palatability with white granulated sugar;
(8) can;
(9) sterilization.
Embodiment 4:
On October 18th, 2003 was 031018 drink 300000ml according to optimization ratio preparation lot number:
(1) separation of bacterial classification is cultivated
Under aseptic, room temperature condition, will be from " hiding clever mushroom " bacterium among the people through sterilization PBS washing three times, collude with platinum ear circle and to get its deep colony, be inoculated on the isolation medium, put then in 37 ℃ of constant incubators, through cultivation in 7 days; Select the perusal bacterium colony to be the cheese shape, milky or dark brown, surface smoothing, glossy, the edge is smooth, and microscopic examination is a large amount of mycelia and the independent bacterium colony of spore.Do secondary separation under the aseptic condition, used culture medium is the same, cultivates 5 days in rearmounted 37 ℃ of constant incubators, selects the same independent bacterium colony, is inoculated under the aseptic condition and preserves on the culture medium preservation under 4 ℃ of conditions;
The raw material of isolation medium and proportioning are:
Potato soup 200ml glucose 15g peptone 10g yeast extract 2g agar 20g adds water to 1000ml
The raw material and the proportioning of preserving culture medium are:
Potato soup 200ml glucose 7g peptone 10g agar 20g adds water to 1000ml
2) preparation of fermented bacterium liquid
The fermented bacterium liquid culture medium uses following raw material and proportioning preparation:
Potato soup 300ml glucose 20g peptone 10g maltose 2g adds water to 1000ml
1. I level fermented bacterium liquid: will be stored in 4 ℃ of kluyveromyces marxianus bacterial classifications under the condition, be inoculated under the aseptic condition in the fermented bacterium liquid culture medium test tube, cultivate 2 days in 30 ℃ of constant incubators, I level fermented bacterium liquid;
2. II level fermented bacterium liquid: with I level fermented bacterium liquid, be inoculated under the aseptic condition in the bottled fermented bacterium liquid culture medium of 250ML, under 30 ℃ of constant temperatures, cultivated 2 days;
(3) first preparation of fermentation liquid:
With II level strain liquid, be inoculated under the aseptic condition in the zymotic fluid culture medium, standing for fermentation is 3 days under 30 ℃ of conditions; Raw materials used and the proportioning of its culture medium is:
Potato soup 195000ml
Glucose 19500g
Maltose 400g
Lactose 200g
Starch sugar 400g
Carrot juice 3000ml
Water 102000ml
(4) original fermentation liquor: the first zymotic fluid that (3) are made is being lower than under 20 ℃ of conditions, and standing for fermentation three days gets drink stoste;
(5) filter;
(6) active ingredient check;
(7) seasoning transfers to sour and sweet palatability with white granulated sugar;
(8) can;
(9) sterilization.
Claims (4)
1, a kind of Marx's Crewe dimension fermented beverage, it is characterized in that adopting a strain kluyveromyces marxianus bacterium-Kluyveromyces marxianus, under aseptic condition, be inoculated in the fermentation broth, raw materials used and the proportioning of described culture medium is: potato soup 50-70 part, 5 parts of glucose, maltose 0.1-0.5 part, 0.1 part of lactose, starch sugar 2-5 part, carrot juice 3-5 part also add water to 1000 parts, standing for fermentation is three days under 25-30 ℃ of condition, makes zymotic fluid just; Just zymotic fluid is being lower than under 20 ℃ of conditions, and standing for fermentation three days is made through aseptic filtration and follow-up conventional treatment then.
2, a kind of preparation method of Marx's Crewe dimension fermented beverage according to claim 1 is characterized in that it is according to following method and process preparation:
The separation of A bacterial classification is cultivated
Under aseptic, room temperature condition, will be from " hiding clever mushroom " bacterium among the people through sterilization PBS washing three times, collude with platinum ear circle and to get its deep colony, be inoculated on the isolation medium, put then in 37 ℃ of constant incubators, through cultivation in 5-7 days; Select the perusal bacterium colony to be the cheese shape, milky or dark brown, surface smoothing is glossy, the edge is smooth, microscopic examination is a large amount of mycelia and the independent bacterium colony of spore, does secondary separation under the aseptic condition, and used culture medium is the same, cultivated 3-5 days in rearmounted 37 ℃ of constant incubators, select the same independent bacterium colony, be inoculated under the aseptic condition and preserve on the culture medium preservation under 4 ℃ of conditions;
Above-mentioned isolation medium is by following raw material and proportioning preparation:
10 parts of 200 parts of glucose 10-15 of potato soup part peptones
2 parts of agar of yeast extract add water to 1000 parts for 20 parts
Above-mentioned preservation culture medium is by following raw material and proportioning preparation:
10 parts of 200 parts of glucose 5-7 of potato soup part peptones
Agar adds water to 1000 parts for 20 parts
The preparation of B fermented bacterium liquid
The fermented bacterium liquid culture medium uses following raw material and proportioning preparation:
Potato soup 200-300 part glucose 15-20 part peptone 10-20 part
Maltose 2-10 part adds water to 1000 parts
1. I level fermented bacterium liquid: will be stored in 4 ℃ of kluyveromyces marxianus bacterial classifications under the condition, be inoculated under the aseptic condition in the fermented bacterium fluid nutrient medium test tube, cultivate 2-3 days in the 25-30 ℃ of constant incubator, I level fermented bacterium liquid;
2. II level fermented bacterium liquid: with I level fermented bacterium liquid, be inoculated under the aseptic condition in the bottled fermented bacterium fluid nutrient medium of 250ML, under 25-30 ℃ of constant temperature, cultivated 2-3 days;
C is preparation of fermentation liquid just:
With II level strain liquid, be inoculated under the aseptic condition in the zymotic fluid corporal punishment culture medium, standing for fermentation is three days under the 25-30 ℃ of condition; Raw materials used and the proportioning of its culture medium is:
0.1 part of 5 parts of maltose 0.1-0.5 of potato soup 50-70 part glucose part lactose
Starch sugar 2-5 part carrot juice 3-5 part adds water to 1000 parts
The preparation of D original fermentation liquor:
The first zymotic fluid that C is made is being lower than under 20 ℃ of conditions, and standing for fermentation three days gets drink stoste;
E filters;
The G seasoning;
The H can;
The J sterilization.
3, a kind of preparation according to claim 2 method of Marx's Crewe dimension fermented beverage according to claim 1, the preparation method who it is characterized in that described separation, preserves culture medium is:
Potato is cleaned peeling, be cut into small pieces, add water, boiled filtered through gauze 30-50 minute in 1: 4 ratio, the all liq of squeezing out, moisturizing adds other batchings respectively to 1000ml, fully dissolving, the accent pH value is 5.5-5.7, packing test tube, 120 ℃ of 30 minutes autoclavings, bevel.
4, a kind of preparation according to claim 2 method of Marx's Crewe dimension fermented beverage according to claim 1 is characterized in that the preparation method of described fermentation broth and fermented bacterium fluid nutrient medium is:
Potato is cleaned peeling, be cut into small pieces, add water, boiled 30-50 minute in 1: 4 ratio, filtered through gauze, all liq of squeezing out, moisturizing adds other batchings respectively to 1000ml, fully dissolving, the accent pH value is 5.5-5.7, packing, 120 ℃ of 30 minutes autoclavings are standby.
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| CNB2003101177462A CN1315417C (en) | 2003-12-30 | 2003-12-30 | Kluyveromyces marxianus yeast fermented drink and its preparation method |
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| CNB2003101177462A CN1315417C (en) | 2003-12-30 | 2003-12-30 | Kluyveromyces marxianus yeast fermented drink and its preparation method |
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| CN1315417C true CN1315417C (en) | 2007-05-16 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1088064A (en) * | 1992-12-12 | 1994-06-22 | 齐齐哈尔轻工学院 | The preparation technology of edible strain fermentation non-alcoholic beverage |
| CN1148479A (en) * | 1996-08-12 | 1997-04-30 | 马永强 | Cereal-bean mixed culture lactic acid fermented drink and preparing method thereof |
| CN1296073A (en) * | 1999-11-16 | 2001-05-23 | 遵义天楼野木瓜有限公司 | Zymophyte and wild oblonga fermented beverage prepared from same, and preparation process thereof |
| US6641856B1 (en) * | 1997-06-13 | 2003-11-04 | Kikue Hashimoto | Beverage containing a kayu-like fermentation product |
-
2003
- 2003-12-30 CN CNB2003101177462A patent/CN1315417C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1088064A (en) * | 1992-12-12 | 1994-06-22 | 齐齐哈尔轻工学院 | The preparation technology of edible strain fermentation non-alcoholic beverage |
| CN1148479A (en) * | 1996-08-12 | 1997-04-30 | 马永强 | Cereal-bean mixed culture lactic acid fermented drink and preparing method thereof |
| US6641856B1 (en) * | 1997-06-13 | 2003-11-04 | Kikue Hashimoto | Beverage containing a kayu-like fermentation product |
| CN1296073A (en) * | 1999-11-16 | 2001-05-23 | 遵义天楼野木瓜有限公司 | Zymophyte and wild oblonga fermented beverage prepared from same, and preparation process thereof |
Non-Patent Citations (1)
| Title |
|---|
| 《西藏干酪乳酸菌降胆固醇特性研究》 肖琳琳，《食品科学》，第24卷第10期 2003 * |
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