WO2018049853A1

WO2018049853A1 - Lactobacillusplantarum proliferator, and fermented product added with the proliferator and preparation method thereof

Info

Publication number: WO2018049853A1
Application number: PCT/CN2017/087523
Authority: WO
Inventors: 张红发; 游春苹; 刘振民
Original assignee: 光明乳业股份有限公司
Priority date: 2016-09-14
Filing date: 2017-06-08
Publication date: 2018-03-22
Also published as: GB201903875D0; GB2568843A; GB2568843B; JP2018530307A; JP6858126B2

Abstract

A Lactobacillus plantarum proliferator is a proteolytic mixture obtained by using a protease or a substance containing a protease to perform a degradation operation on a plant protein. The protease is one of a plant protease, an animal protease, a microbial protease, or a mixture thereof. The plant protein having undergone the degradation operation is one of a soybean protein, a mung bean protein, a chickpea protein, a broad bean protein, a pea protein, a hyacinth bean protein, a wheat protein, or a protein mixture thereof. A fermented product added with the Lactobacillus plantarum proliferator contains viable Lactobacillus plantarum of 1.0×10⁹ cfu/g. A method of preparing the fermented product comprises the following steps: (1) mixing a main ingredient with Lactobacillus plantarum; (2) performing sterilization; and (3) performing inoculation and fermentation.

Description

Lactobacillus plantarum proliferation agent, fermentation product added with the same, and preparation method thereof

Technical field

The present invention belongs to the field of microbial fermentation and food processing technology, in particular, in a fermented food, a Lactobacillus plantarum proliferator which plays an important role in increasing the number of live bacteria of Lactobacillus plantarum, and a fermented product produced by using the proliferator And its preparation method.

Background technique

Probiotics are a kind of active microorganisms beneficial to the host. They are the general term for active beneficial microorganisms that colonize the human intestines and reproductive system and produce precise health effects to improve the balance of the host micro-ecology and play a beneficial role. At present, the most powerful products studied in the world are mainly compound active probiotics composed of various microorganisms, which are widely used in bioengineering, industrial and agricultural, food safety and life health fields, and their probiotic functions are increasingly favored by consumers. Identity and welcome. Lactobacillus plantarum is one of the probiotics, which can effectively tolerate the human digestive system, regulate intestinal flora balance, promote digestion and absorption, regulate cholesterol, regulate blood lipids, enhance human immunity, prevent cancer and inhibit tumor growth. Probiotic health function. Therefore, the development of high viable bacterial counts of Lactobacillus plantarum products is of positive significance for improving consumer health. At present, the probiotic products on the market are mainly Streptococcus thermophilus and Lactobacillus bulgaricus as the main starter, and other functional probiotics are added at the same time. The pure probiotic fermentation products are few and far, and all the live bacteria products are not carried out. Detection of bacteria.

Because Lactobacillus plantarum grows slowly in cow's milk, it has poor acid-producing ability and is difficult to curd. There are many studies at home and abroad to try to solve this problem, but there are some defects, and actual production is difficult. Paper "Study on Growth Limiting Factors of Lactobacillus Plantarum ST-III in Skim Milk System" [Chen Shuai et al., Food and Fermentation Industry, 2008, 34(10): 53-57] and "Efficient Exogenous Additive Fish Peptone Group" Study on the fractionation of fractions [Chen Shuai et al., Dairy Science and Technology, 2010, 4:201-204] studied the promotion of fermentation of Lactobacillus plantarum in milk by different protein components in fish peptone, but the fish used. Peptone is reagent grade; at the same time, the fermentation time is longer (24h), the cost is high, and it is difficult to meet the actual product production requirements. Patent "an active plant Lactobacillus drink and its preparation method" [Ma Chengjie et al, CN102715235A: 2012-10-10] discloses an active plant Lactobacillus proliferator formulation, planted Lactobacillus has a high live content. However, the reagents used are complex in composition and the fermentation time is longer (24-72h). Patent "a Lactobacillus plantarum fermented milk and preparation method thereof" [Ma Chengjie et al, CN103004984A: 2013-04-03] discloses a Lactobacillus plantarum fermented milk and a preparation method thereof, mainly adding an appropriate amount of amino acids and nucleotides, The limiting factor for the growth and reproduction of Lactobacillus plantarum in milk was removed. However, the additive composition is complicated and the cost is high; and the fermentation time of the pure Lactobacillus plantarum is longer (15h). The paper "Optimization of fermentation process of Lactobacillus plantarum ST-III skim milk" [Hua Baozhen et al., Journal of Agricultural Engineering, 2014, 30 (23711): 276-284] studied the proliferation of soybean polypeptide by Lactobacillus plantarum, live bacteria The number is higher; however, the fermentation time is longer (48h).

Foreign studies on Lactobacillus plantarum are less proliferating. Among them, a Japanese company uses the grain distiller's grains extract to proliferate Lactobacillus plantarum, but only has a proliferative effect on some Lactobacillus plantarum, and the fermentation time is longer (more than 16h), especially for some The proliferation effect of Lactobacillus plantarum (such as Lactobacillus plantarum ST-III) is not satisfactory. This is mainly because the Lactobacillus plantarum genes have large differences, and the nutritional needs of different strains are different.

Both domestic and foreign studies have neglected the detection of contamination in the pure fermentation products of Lactobacillus plantarum. If the proliferation method studied has the characteristics of short fermentation time and high viable count, it is meaningless to detect no bacteria. Everyone knows that milk and common foods often contain heat-resistant Bacillus contamination, which is difficult to kill. If Lactobacillus plantarum does not grow as soon as possible to inhibit the growth of bacteria, the bacteria in the fermented milk will greatly exceed the standard; especially some harmful bacteria such as wax Bacillus, staphylococcus, etc. will seriously endanger human health. This requires that the proliferation method of the Lactobacillus plantarum studied not only has a good enrichment effect, but also has a high number of viable bacteria; and it is required to grow rapidly, thereby inhibiting the growth of the bacteria.

In order to shorten the growth and acid production time of Lactobacillus plantarum in milk, it is a feasible solution to add a proliferative agent during the fermentation process. Exploring the composition of the proliferator is the key to increasing the activity of Lactobacillus plantarum (increasing the number of viable bacteria) and reducing the fermentation time to achieve control of harmful bacteria. Fermented products made by Lactobacillus plantarum proliferator are first required to have higher requirements on the number of live bacteria in the product, and at the same time, the fermentation time is required to be short, and the content of other harmful bacteria is reduced as much as possible; in addition, the number of viable bacteria in the preservation period is Higher to ensure the long-term effectiveness of its products. In the field of food industry, research on high-efficiency Lactobacillus multiplication agents has become the focus and frontier of biotechnology research in this field of technology.

Summary of the invention

The technical problem to be solved by the technical solution of the present application is to propose a novel proliferation method for the existing Lactobacillus plantarum multiplication agent having a proliferative effect on some specific Lactobacillus plantarum, and the fermentation time is long and the proliferation effect is not obvious. Agent. The addition of the proliferator can significantly increase the content of Lactobacillus plantarum, the fermentation time is short, the mouthfeel is good, the contamination of the bacteria is reduced, and the technical problem of fermenting the live bacteria product is solved. At the same time, the fermenting agent is used to improve various fermented foods, and a fermented food having a high content of Lactobacillus plantarum and a preparation method thereof are proposed.

The technical solution for realizing the above-mentioned object of the invention is: a Lactobacillus plantarum multiplication agent which is an enzymatic hydrolysis mixture after degrading a vegetable protein by a protease or a protease-containing substance. The protease is one or a mixture of a vegetable protease, an animal protease or a microbial protease; the protease-containing substance is one or a mixture of a substance containing a plant protease, a substance of an animal protease or a substance of a microbial protease.

The technical solution for adding the Lactobacillus plantarum multiplication agent fermentation product is: a fermentation product to which a Lactobacillus plantarum multiplication agent is added, wherein the Lactobacillus plantarum proliferator is degraded by a protease or a protease-containing substance The enzymatically hydrolyzed mixture contains Lactobacillus plantarum proliferator in a mass percentage of 0.1-20%. The protease in the above Lactobacillus plantarum multiplying agent is one or a mixture of a plant protease, an animal protease or a microbial protease; and the protease-containing substance is one or a mixture of a substance containing a plant protease, a substance of an animal protease or a substance of a microbial protease. The fermented product is fermented milk or live bacteria drinking water or live bacteria cosmetics.

The vegetable protein may be selected from soy protein, mung bean protein, chickpea protein, pea protein, broad bean protein, lentil protein, wheat protein or a mixture of the above proteins. The above plant proteases usually select fruit protease, vegetable protease, medicinal plant protease (such as mogroside protease, yam protease) or a mixture thereof. The substance containing these plant proteases is a fruit, vegetable or medicinal plant. If an animal protease is selected, it may specifically be trypsin, pepsin, chymotrypsin or a mixture thereof.

If microbial protease is selected, it can be specifically Bacillus licheniformis and Aspergillus niger. Protease, Bacillus amyloliquefaciens, Bacillus subtilis, M. melilase, Aspergillus protease, Kluyveromyces protease, Trichoderma protease, Aspergillus foci, and parasitic internal capsid (P. pallidum) protease One or a mixture thereof.

The present invention also proposes that the Lactobacillus plantarum multiplication agent can reach 1.0×10 ⁹ cfu/g in increasing the live bacteria of the Lactobacillus plantarum.

The steps of the fermented milk production method of the above fermented product are: (1) milk and Lactobacillus plantarum multiplying agent are mixed according to a mass ratio of 100:0.1-20; (2) sterilized and cooled to 37 ° C; (3) inoculation to Lactobacillus plantarum The final concentration is 1.0×10 ⁵ -1.0×10 ⁸ cfu/g; (3) Fermentation to curd, the fermentation time does not exceed 12 h.

The steps for preparing the above-mentioned living bacteria drinking water are as follows: (1) pure water and Lactobacillus plantarum multiplying agent are mixed according to a mass ratio of 100:0.1-20; (2) pre-heating of the pre-mixed liquid is achieved at 65 ° C and 20 MPa. Quality, sterilization, cooling to 37 ° C; (4) inoculation, fermentation for 16h to reach a final concentration of Lactobacillus plantarum of 2.0 × 10 ⁹ cfu / g; (5) in the product of step (4) according to 200mg / 100ml added vitamin C And diluted with purified water to a final concentration of Lactobacillus plantarum 2.0 × 10 ⁸ cfu / g.

The method for preparing the above-mentioned living mushroom cosmetic is as follows: (1) mixing the cosmetic with the Lactobacillus plantarum multiplying agent at a mass ratio of 100:0.1-20; (2) sterilizing; (3) inoculating and fermenting.

The Lactobacillus plantarum multiplication agent and the fermented product thereof provided by the technical solution of the present application have the following advantages: (1) the proliferative agent belongs to a natural edible mixture, and does not cause side effects to the human body; (2) the proliferator shortens the fermentation in use. Time, thus avoiding the contamination of harmful bacteria; (3) using the proliferator to achieve the growth of Lactobacillus plantarum can reach 1.0 × 10 ⁹ cfu / g, the number of viable bacteria is much higher than that produced by other proliferators The number of viable bacteria; (4) The use cost of the proliferator is low, and the production cost of the Lactobacillus plantarum product is lowered.

detailed description

In order to explain the technical solution of the present invention more clearly, the technical solutions of the present application are specifically described below in conjunction with the experimental test results. The Lactobacillus plantarum proliferator uses a protease to degrade the enzymatic hydrolysis mixture produced by the vegetable protein; the vegetable protein can be selected from soybean protein or mung bean protein, chickpea protein, pea protein, broad bean protein, lentil protein, wheat protein or the above protein. mixture. The above plant proteases usually select fruit protease, vegetable protease, medicinal plant protease (such as mogroside protease, yam protease) or a mixture thereof. The substance containing these plant proteases is a fruit, vegetable or medicinal plant. If animal protease is selected, it may specifically be trypsin, pepsin or chymotrypsin.

If microbial protease is selected, it may specifically be Bacillus licheniformis, Aspergillus niger protease, Bacillus amyloliquefaciens, Bacillus subtilis protease, M. melilase, Aspergillus oryzae protease, Kluyveromyces lactis, Trichosporin , one or a mixture of a honey aspergillus protease, a parasitic inner shell (Chestnut bacillus) protease.

The above Lactobacillus plantarum multiplying agent is prepared by enzymatically reacting vegetable protein and vegetable protease in an aqueous solution at 25 ° C - 80 ° C for 0.5-8 h.

Example 1:

In order to determine the action of the proliferator, 0.5-10% of the Lactobacillus plantarum proliferator prepared above was added to milk, sterilized at 95 ° C for 10 min, and the final concentration was 1.0×10 ⁵ -1.0×10 ⁸ cfu. / g Lactobacillus plantarum ST-III, fermented to curd (not more than 12h), the total number of aerobic colonies was counted by LB plate to detect the contamination, and the number of live bacteria of Lactobacillus plantarum was counted by MRS plate. The curd test results obtained by the method are shown in Table 1:

Table 1. Results of pure fermented milk of Lactobacillus plantarum

The experimental results in Table 1 prove that the proliferative agent can promote the rapid growth of curd of Lactobacillus plantarum ST-III in milk, and the number of viable cells in the 30-day shelf life does not change much, no bacteria contamination; and the number of Lactobacillus plantarum viable counts Higher.

Example 2:

The soy protein and the ficin protease are used for enzymatic hydrolysis to form a proliferative agent, and the proliferative agent is mixed with the raw milk, inoculated, fermented, and the curd condition is observed. The sample is diluted by the sample, and the sample is refrigerated for 30 days and then counted; The effect was tested by using soy peptone and tryptone as a proliferation agent. The results are shown in Table 2.

Table 2. Results of controlled trials of soy peptone and tryptone as proliferator

增殖剂Proliferator	凝乳情况Curd condition	好氧菌落总数(cfu/g)Total aerobic colonies (cfu/g)	植物乳杆菌活菌计数(cfu/g)Lactobacillus plantarum viable count (cfu/g)
本技术方案的增殖剂Proliferator of the technical solution	凝乳curd	<1×10¹ <1×10 ¹	1.2×10⁹ 1.2×10 ⁹
大豆蛋白胨Soy protein	未凝乳Uncured	2.1×10³ 2.1×10 ³	3.1×10⁸ 3.1×10 ⁸
胰蛋白胨Tryptone	未凝乳Uncured	2.1×10⁵ 2.1×10 ⁵	1.5×10⁸ 1.5×10 ⁸

It can be seen from Table 2 that the enrichment effect of the enrichment agent of the technical solution of the present application far exceeds the effect of soy peptone and tryptone.

Example 3:

Using a method similar to the above-described examples, the enzymatic hydrolysis reaction was carried out using soy protein and bromelain to produce a proliferative agent, and the coagulation was observed by mixing, fermenting, and inoculation with the proliferative agent and raw milk, and the results are shown in Table 3.

Table 3. Experimental results of enzymatic hydrolysis of soy protein and bromelain to produce a proliferator

时间time	凝乳情况Curd condition	好氧菌落总数(cfu/g)Total aerobic colonies (cfu/g)	植物乳杆菌活菌计数(cfu/g)Lactobacillus plantarum viable count (cfu/g)
0天0 days	凝乳curd	<1×10¹ <1×10 ¹	1.4×10⁹ 1.4×10 ⁹
30天后30 days later	凝乳curd	<1×10¹ <1×10 ¹	5.0×10⁸ 5.0×10 ⁸

Example 4:

The above-mentioned soy protein and pineapple whole juice were subjected to enzymatic hydrolysis to form a proliferative agent, and the coagulant was observed by mixing, fermenting, and inoculating with the proliferative agent and raw milk, as shown in Table 4.

Table 4. Experimental results of enzymatic hydrolysis of soy protein and pineapple whole juice to produce proliferator

时间time	凝乳情况Curd condition	好氧菌落总数(cfu/g)Total aerobic colonies (cfu/g)	植物乳杆菌活菌计数(cfu/g)Lactobacillus plantarum viable count (cfu/g)
0天0 days	凝乳curd	<1×10¹ <1×10 ¹	1.0×10⁹ 1.0×10 ⁹
30天后30 days later	凝乳curd	<1×10¹ <1×10 ¹	4.5×10⁸ 4.5×10 ⁸

Example 5:

The test results of the enzymatic hydrolysis reaction using soybean protein and the following plant proteases to prepare a multiplying agent are shown in Table 5 below.

Table 5. Effect of different plant protease extracts

植物蛋白酶Plant protease	凝乳情况Curd condition	好氧菌落总数(cfu/g)Total aerobic colonies (cfu/g)	植物乳杆菌活菌计数(cfu/g)Lactobacillus plantarum viable count (cfu/g)
木瓜蛋白酶papain	凝乳curd	<1×10¹ <1×10 ¹	1.1×10⁹ 1.1×10 ⁹
无花果蛋白酶Fig protease	凝乳curd	<1×10¹ <1×10 ¹	1.0×10⁹ 1.0×10 ⁹
菠萝蛋白酶Bromelain	凝乳curd	<1×10¹ <1×10 ¹	9.5×10⁸ 9.5×10 ⁸
罗汉果蛋白酶Mogroside protease	凝乳curd	<1×10¹ <1×10 ¹	9.6×10⁸ 9.6×10 ⁸

Example 6

The above soy protein can also be replaced with other vegetable proteins, which are enzymatically hydrolyzed with pineapple juice. The proliferation was prepared by the reaction and the test results are shown in Table 6 below.

Table 6. Effect of proliferation agent enrichment experiment on enzymatic hydrolysis of different plant proteins and pineapple juice

Example 7

The above plant protease can also be replaced with an animal protease, which is subjected to enzymatic hydrolysis with soybean protein to prepare a proliferation agent, and tested, and the results are shown in Table 7 below.

Table 7. Effect of different animal protease enrichment experiments

蛋白酶Protease	凝乳情况Curd condition	好氧菌落总数(cfu/g)Total aerobic colonies (cfu/g)	植物乳杆菌活菌计数(cfu/g)Lactobacillus plantarum viable count (cfu/g)
胰蛋白酶Trypsin	凝乳curd	<1×10¹ <1×10 ¹	9.2×10⁸ 9.2×10 ⁸
胃蛋白酶Pepsin	凝乳curd	<1×10¹ <1×10 ¹	9.6×10⁹ 9.6×10 ⁹
胰凝乳蛋白酶Chymotrypsin	凝乳curd	<1×10¹ <1×10 ¹	8.3×10⁸ 8.3×10 ⁸

Example 8

The above plant protease can also be replaced with a microbial protease, which is subjected to enzymatic hydrolysis with soybean protein to prepare a proliferator, and tested. The results are shown in Table 8 below.

Table 8. Different microbial protease enrichment experiments

Example 9

Lactobacillus plantarum proliferator is soybean + pineapple juice

Lactobacillus plantarum ST-III can also be replaced with other Lactobacillus plantarum strains.

Table 9. Different Lactobacillus plantarum proliferation assays

A variety of fermented products having a high viable cell count can be produced by using the above-mentioned Lactobacillus plantarum multiplying agent, and the production processes of several fermented products are listed below in Examples 10-18.

Example 10: Lactobacillus plantarum fermented milk

Lactobacillus plantarum fermented milk formula: 80% raw milk, 2% of proliferator (solid content), appropriate amount of thickener, 7% sucrose, 5% orange juice.

The preparation method comprises the following steps: preheating and heating the mixture liquid, homogenizing at 65° C. and 20 MPa, sterilizing at 95° C. for 10 minutes, and cooling to 37° C. Inoculation (final concentration of ST-III inoculum 1.0 × 10 ⁶ cfu/g) was fermented to curd. Results The number of viable bacteria of Lactobacillus plantarum was 1.0×10 ⁹ cfu/g, and the number of viable cells in the 21-day preservation period was more than 3.0×10 ⁸ cfu/g, which was sweet and sour. At present, there is no pure Lactobacillus fermented milk product in the domestic and foreign markets.

Example 11: Lactobacillus plantarum instant fermented milk powder

The Lactobacillus plantarum fermented milk obtained in Example 10 was first spray-dried to a water content of about 20% powder. The spray-dried inlet air temperature is 110 ° C, the spray-dried exhaust air temperature is 37 ° C, the powder outlet temperature is 35 ° C; and then fluidized and dried in a fluidized bed at a low temperature of 50 ° C. A fermented milk powder having a water content of <5% in the powder. The method for using the fermented milk powder fermented yoghurt comprises the steps of: mixing the fermented milk powder prepared above with a mass ratio of warm water (50 ° C) of 1:4, and maintaining the instant fermented milk for half an hour. The number of viable bacteria of Lactobacillus plantarum was 3.0 × 10 ⁸ cfu / g. At present, there is no Lactobacillus plantarum instant fermented milk product on the domestic and international markets. Example 12: Lactobacillus planta yoghurt ice cream

Raw materials: raw milk 40%, proliferative agent 6% (solid content), white sugar 10%, skim milk powder 10%, cream 20%, stabilizer 4% (main ingredient is sodium carboxymethyl cellulose, starch) All the raw materials were mixed, homogenized at 65 ° C, and sterilized at 99 ° C for 10 min; cooled to 37 ° C for inoculation (final concentration of ST-III inoculum 1.0 × 10 ⁷ cfu / g) for 12 h. The fermentation product was stirred for 20 min at 45 ° C and 150 rpm, and cooled to 15 ° C for 16 h. O ° C was allowed to stand for aging for 12 h, and then homogenized under a pressure of 40 ° C. Finally, put the ice cream machine at -30 ° C to make yogurt ice cream. The obtained product is sweet and sour, and has a probiotic function of Lactobacillus plantarum (the number of viable cells within 3 months > 4.0 × 10 ⁸ cfu / g). There are currently no Lactobacillus plantarum yogurt ice cream products on the domestic and international markets.

Example 13: Lactobacillus plantarum live drink

(1) Fermented milk base formula: 15% skim milk powder, 4% proliferation agent (solid content), appropriate amount of thickener, 7% sucrose, juice 10%. Fermented milk base preparation method: preheating and heating the feed liquid, after homogenization, heat sterilization at 105 ° C for 10 min, and cooling to 37 ° C. Inoculation (final concentration of ST-III inoculum 1.0 × 10 ⁶ cfu/g) was fermented to curd.

(2) Production of live mushroom type yogurt drink

Preparation: white sugar 10%, water 45%, stabilizer (pectin) 0.5%, will be stirred according to the above ratio of stabilizer and an appropriate amount of white sugar, water, high pressure homogenization, sterilization and cooling. Add about 40% of the fermented milk base, stir and mix, adjust the acid to pH 4.4, and adjust the fragrance. Sterile homogenization at 20 MPa to obtain a Lactobacillus plantarum live bacterial beverage. After refrigerating at 4 ° C for 1 month, the number of viable bacteria of Lactobacillus plantarum was >1.0×10 ⁸ cfu/g. At present, there are few live Lactobacillus plantarum beverages on the market, generally using direct injection bacteria powder, the cost is high, and the number of viable bacteria is small, only 2.0×10 ⁷ cfu/g.

Example 14: Lactobacillus plantarum live bacteria water

Lactobacillus plantarum high-density fermented milk formula: 10% skim milk powder, 5% proliferation agent (solid content).

Preparation method: preheating the feed liquid, homogenizing at 65 ° C, 20 MPa, sterilizing at 115 ° C for 10 min, and cooling to 37 ° C. The inoculation (final concentration of ST-III inoculum was 5.0×10 ⁶ cfu/g), and the number of viable bacteria of Lactobacillus plantarum was 2.0×10 ⁹ cfu/g after fermentation for 16 hours. The diluted water was added to purified water containing 200 mg/100 ml of vitamin C and 0.2% of a pectin stabilizer to prepare a living bacteria water having a final concentration of 2.0 × 10 ⁸ cfu/g. At present, there is no Lactobacillus plantarum live aquatic product in the domestic and foreign markets.

Example 15: Lactobacillus plantarum probiotic preparation

Lactobacillus plantarum high-density fermented milk formula: 12% skim milk powder, 5% proliferation agent (solid content).

Preparation method: preheating the feed liquid, homogenizing at 65 ° C, 20 MPa, sterilizing at 115 ° C for 10 min, and cooling to 37 ° C. The inoculation (final concentration of ST-III inoculum was 2.0×10 ⁷ cfu/g), and the number of viable bacteria of Lactobacillus plantarum was 1.5×10 ⁹ cfu/g after fermentation for 12 hours. The fermentation broth was centrifuged, and 5% sucrose, L-cysteine 0.01%, and the Lactobacillus plantarum powder obtained by vacuum drying or freeze-drying were added to the slime. The Lactobacillus plantarum powder obtained by vacuum drying is mixed with lactose after grinding, and is tableted or made into a powder capsule. The viable cell count was 1.0×10 ¹⁰ cfu/g, and the survival rate after drying at room temperature for one year was more than 40%. The medium used in the probiotics preparation of Lactobacillus plantarum on the market is inedible, and it is necessary to separate the cells, and the production cost is high.

Example 16: Lactobacillus plantarum mixed fermented milk

Lactobacillus plantarum fermented milk formula: 80% raw milk, 2% of proliferator (solid content), appropriate amount of thickener, 7% sucrose.

Preparation method: preheating the feed liquid, homogenizing at 65 ° C, 20 MPa, sterilizing at 90 ° C for 8 min, and cooling to 37 ° C. Inoculation (final concentration of ST-III inoculum 2.0×10 ⁶ cfu/g, Lactobacillus bulgaricus powder 0.02 g/kg, Streptococcus thermophilus powder 0.03 g/kg), fermented to curd, with or without added fruit material such as: Strawberries, kiwis, etc. Results The number of viable bacteria of Lactobacillus plantarum was 4.0×10 ⁸ cfu/g, and the number of viable cells in the 21-day preservation period was more than 2.0×10 ⁸ cfu/g, which was sweet and sour. At present, some fermented milk on the market contains Lactobacillus plantarum, which is generally direct injection of bacteria powder, and has high cost and low viable count (close to 1.0×10 ⁷ cfu/g.).

Example 17: Lactobacillus plantarum lactic acid bacteria beverage

Fermented milk base: The fermented milk produced in Example 10 was used as a fermented milk base.

Preparation: white sugar 10%, water 45%, stabilizer (pectin) 0.5%, will be stirred according to the above ratio of stabilizer and appropriate amount of white sugar, water, high pressure homogenization, sterilization and cooling. Add the fermented milk base (40%), stir and mix, adjust the acid to pH 4.4, and adjust the fragrance. Sterile homogenization at 20 MPa to obtain a Lactobacillus plantarum live bacterial beverage. After refrigerating at 4 ° C for 1 month, the number of viable bacteria of Lactobacillus plantarum was >1.0×10 ⁸ cfu/g. Since Lactobacillus plantarum is difficult to proliferate in milk, the Lactobacillus plantarum lactic acid bacteria beverage currently on the market adopts direct injection of bacteria powder, has high cost, and has a low viable cell count of less than 1.0×10 ⁷ cfu/g.

Example 18: Living bacteria cosmetic containing Lactobacillus plantarum

Formulation 1: A Vaseline glycerin moisturizer was used as a substrate, and the Lactobacillus plantarum viable bacteria prepared in Example 1 was mixed at 100:1-20. Applying to the skin surface, it has the effect of inhibiting the growth of bacteria and acne.

Formulation 2: A mask material was used as a substrate (e.g., beeswax, sodium hyaluronate, collagen), and the Lactobacillus plantarum viable bacteria prepared in Example 1 was mixed at 100:1-20. Applying to the skin surface, it has the effect of inhibiting the growth of bacteria and acne.

The above is only the preferred embodiment of the present invention, and is not intended to limit the present invention. Any modifications, equivalent substitutions, and simple modifications made in the substance of the present invention are included in the scope of the present invention. Inside.

Claims

A Lactobacillus plantarum multiplying agent characterized in that the Lactobacillus plantarum multiplying agent is an enzymatic mixture after degrading a vegetable protein by a protease or a protease-containing substance.
The Lactobacillus plantarum multiplying agent according to claim 1, wherein the protease is one of a plant protease, an animal protease or a microbial protease or a mixture thereof.
The Lactobacillus plantarum multiplying agent according to claim 1, wherein the protease-containing substance is one of a substance containing a plant protease, a substance of an animal protease, or a substance of a microbial protease, or a mixture thereof.
The Lactobacillus plantarum multiplication agent according to claim 1, wherein the vegetable protein for degradation is one of soybean protein, mung bean protein, chickpea protein, broad bean protein, pea protein, lentil protein, and wheat protein. Or a mixed protein thereof.
The Lactobacillus plantarum multiplying agent according to claim 1, wherein the protease or the protease-containing substance degrades the vegetable protein reaction in a mass ratio of 1:1 to 50.
The Lactobacillus plantarum multiplying agent according to claim 2, wherein the plant protease is a fruit protease, a vegetable protease, a mogroside protease, a yam protease, or a mixture thereof.
The Lactobacillus plantarum multiplying agent according to claim 2, wherein the animal protease is one of trypsin, pepsin, chymotrypsin or a mixture thereof.
The Lactobacillus plantarum multiplication agent according to claim 2, wherein the microbial protease is Bacillus licheniformis, Aspergillus niger protease, B. amyloliquefaciens, Bacillus subtilis protease, Rhizomucor miehei protease, Aspergillus protease, One of a Kluyveromyces lactis protease, a micromycin protease, a honey Aspergillus protease, a parasitic internal capsid (P. infestans) protease, or a mixture thereof.
The Lactobacillus plantarum multiplying agent according to claim 3, wherein the plant protease-containing substance is one of a fruit, a vegetable, a medicinal plant or a mixture thereof.
A fermented product to which the Lactobacillus plantarum multiplying agent according to any one of claims 1 to 9 is added, characterized in that the Lactobacillus plantarum viable bacteria in the fermented product reaches 1.0 × 10 9 cfu/g.
The fermentation product to which a Lactobacillus plantarum multiplication agent is added according to claim 10, wherein the Lactobacillus plantarum multiplication agent is an enzymatic hydrolysis mixture after degrading a vegetable protein by a protease or a protease-containing substance, the product containing the plant The quality of Lactobacillus proliferation agent The fraction is 0.1-20%, and the fermented product is fermented milk or live bacteria drinking water or live bacteria cosmetics.
The fermented product to which a Lactobacillus plantarum multiplication agent is added according to claim 11, wherein when the product is fermented milk, it contains 0.1 to 20% by mass of a Lactobacillus plantarum multiplier.
The fermented product to which Lactobacillus plantarum multiplying agent is added according to claim 11, wherein when the product is a fermented milk beverage, it contains 0.1 to 20% by mass of a Lactobacillus plantarum multiplier.
The fermented product to which Lactobacillus plantarum multiplying agent is added according to claim 11, wherein when the product is a live drinking water, it contains 0.1 to 20% by mass of a Lactobacillus plantarum multiplying agent.
The fermented product to which a Lactobacillus plantarum multiplication agent is added according to claim 11, wherein when the product is a live cosmetic, it contains 0.1 to 20% by mass of a Lactobacillus plantarum multiplying agent.
A method for producing a fermented product of the Lactobacillus plantarum multiplication agent according to claim 11, characterized in that the method for preparing the fermented milk is as follows: (1) milk and Lactobacillus plantarum multiplying agent according to a mass ratio of 100:0.1- 20 mixing; (2) sterilization; (3) inoculation to reach a final concentration of Lactobacillus plantarum 1.0 × 10 5 - 1.0 × 10 8 cfu / g; (3) fermentation to curd, fermentation time does not exceed 12h.
A method for producing a fermented product of the Lactobacillus plantarum multiplication agent according to claim 11, wherein the step of preparing the living bacteria drinking water is as follows: (1) pure water and Lactobacillus plantarum multiplying agent according to a mass ratio of 100 : (0) mixing; (2) pre-mixing liquid preheating at 65 ° C, 20 MPa to achieve homogenization, sterilization, cooling to 37 ° C; (4) inoculation, fermentation 16h to reach the final concentration of Lactobacillus plantarum is 2.0 × 10 9 cfu/g; (5) Vitamin C was added in 200 mg/100 ml in the product of step (4), and diluted with purified water to a final concentration of Lactobacillus plantarum 2.0×10 8 cfu/g.
A method for producing a fermented product of the Lactobacillus plantarum multiplication agent according to claim 11, wherein the method for preparing the viable mushroom cosmetic is as follows: (1) the cosmetic and the Lactobacillus plantarum multiplying agent are in a mass ratio of 100: 0.1-20 mixing; (2) sterilization, cooling to 37 ° C; (3) inoculation, fermentation.