CN112094783B - Composite lactobacillus starter and application thereof - Google Patents
Composite lactobacillus starter and application thereof Download PDFInfo
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- CN112094783B CN112094783B CN202011048958.XA CN202011048958A CN112094783B CN 112094783 B CN112094783 B CN 112094783B CN 202011048958 A CN202011048958 A CN 202011048958A CN 112094783 B CN112094783 B CN 112094783B
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- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/06—Treating tea before extraction; Preparations produced thereby
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Abstract
The invention discloses a composite lactobacillus starter and application thereof, and belongs to the technical field of microorganisms and fermentation. The invention provides a compound lactobacillus starter for fermented tea, which is fully dissolved in purified water and then fermented at 37 ℃, so that the viable count of lactobacillus in the fermented tea beverage obtained by fermentation is 1 multiplied by 10 6 Proliferation of cfu/mL to over 2X 10 9 cfu/mL. The invention provides a fermented tea beverage, which is obtained by fermenting composite lactobacillus starter at constant temperature of 35-40 ℃ for 10-14 h, wherein the viable count of lactobacillus in the fermented tea beverage can be up to 2 x 10 9 cfu/mL or more, good effect of benefiting is achieved.
Description
Technical Field
The invention relates to a composite lactobacillus starter and application thereof, belonging to the technical field of microorganisms and fermentation.
Background
The tea is one of three beverages in the world, is rich in various nutritional ingredients such as tea polyphenol, tea polysaccharide and the like, has high nutritional value and multiple health care effects, and is popular with consumers due to convenient drinking, low toxic and side effects and remarkable effects in regulating abnormal glycolipid metabolism.
The lactobacillus fermented tea beverage is very rare in the market, and the viable bacteria type fermented tea beverage is more blank. While the rise of the Kangpu tea has been attracting interest of some researchers and exploring a composite fermented product of fruit juice and tea, for example, huang Taowu dragon composite fermented tea beverage obtained by fermenting tea with yeast dedicated to fruit wine after adding fruit juice to tea, dark plum Kangpu tea composite beverage obtained by compounding fruit juice concentrate with Kangpu tea, and the like. However, the Kangpu tea is prepared by composite fermentation of multiple strains such as saccharomycetes, acetic acid bacteria and lactic acid bacteria, and the like, and after inactivation, the tea is prepared into tea-containing drinks with various tastes according to requirements, and the complexity and uncontrollability of fermentation strains of the tea-containing drinks have influence on standardization of products.
Lactobacillus is an important probiotic for human body, and has been widely applied to yoghurt, active probiotic drinks (such as yangle) and the like. At present, lactobacillus starter of fermented tea and fermented tea beverage containing active probiotics are still blank. This is mainly because lactobacillus is auxotrophic and hardly proliferates in pure tea systems with poor nutrient content.
Therefore, there is an urgent need to develop a lactobacillus starter culture for tea to produce a fermented tea beverage rich in lactobacillus.
Disclosure of Invention
[ technical problem ]
The invention aims to provide a composite lactobacillus starter for tea which can be fermented.
Technical scheme
In order to solve the technical problem of the invention, the invention provides a composite lactobacillus starter, which contains tea powder and lactobacillus powder; the tea powder is prepared by leaching tea with water to obtain tea soup, adding soluble carbohydrate, proliferation factor, inorganic salt and active protectant into the tea soup to obtain mixed solution, and drying the mixed solution.
In one embodiment of the invention, the tea is one or more of black tea or jasmine tea.
In one embodiment of the invention, the tea leaves are 10-30 per mill of the water mass during leaching.
In one embodiment of the invention, the soluble carbohydrate is one or more of glucose, fructose, sucrose, lactose, maltose or fruit and vegetable juice.
In one embodiment of the invention, the fruit and vegetable juice is one or more of apple juice, pineapple juice or carrot juice.
In one embodiment of the invention, the addition amount of the soluble carbohydrate in the tea soup accounts for 1-8% of the total mass of the tea soup.
In one embodiment of the present invention, the proliferation factor is one or more of yeast extract, yeast protein, yeast peptone, soybean peptone, tryptone, and casein peptone.
In one embodiment of the invention, the addition amount of the proliferation factor in the tea soup is 2-10 per mill of the total mass of the tea soup.
In one embodiment of the present invention, the inorganic salt is one or more of dipotassium hydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate or sodium potassium phosphate;
or the inorganic salt is one or more of sodium citrate or potassium citrate.
In one embodiment of the invention, when the inorganic salt is one or more of dipotassium hydrogen phosphate, disodium hydrogen phosphate, monopotassium phosphate, sodium dihydrogen phosphate, potassium phosphate or sodium potassium phosphate, the adding amount of the inorganic salt in the tea soup accounts for 2-8 per mill of the total mass of the tea soup;
when the inorganic salt is one or more than one of sodium citrate and potassium citrate, the adding amount of the inorganic salt in the tea soup is 9-20 per mill of the total mass of the tea soup.
In one embodiment of the present invention, the active protectant is one or more of whey protein isolate, casein, soy protein, collagen, or skim milk;
alternatively, the active protectant is a polysaccharide;
or the active protective agent is one or more of dietary fiber, resistant dextrin, maltodextrin, polydextrose, oligosaccharide or hydroxyethyl starch.
In one embodiment of the invention, when the active protective agent is one or more than one of whey protein isolate, casein, soy protein, collagen or skim milk, the addition amount of the active protective agent in the tea soup is 2-5% of the total mass of the tea soup;
when the active protective agent is polysaccharide, the addition amount of the active protective agent in the tea soup is 0.1-5% of the total mass of the tea soup;
when the active protective agent is one or more of dietary fiber, resistant dextrin, maltodextrin, polydextrose, oligosaccharide or hydroxyethyl starch, the addition amount of the active protective agent in the tea soup is 0.1-5% of the total mass of the tea soup.
In one embodiment of the invention, the lactobacillus powder is one or more of lactobacillus plantarum powder, lactobacillus casei powder, lactobacillus rhamnosus powder, lactobacillus paracasei powder, lactobacillus bulgaricus powder, lactobacillus sake powder, lactobacillus helveticus powder, lactobacillus salivarius powder, lactobacillus crispatus powder, lactobacillus grignard powder, lactobacillus johnsonii powder, lactobacillus reuteri powder or lactobacillus fermentum powder.
In one embodiment of the present invention, the viable count of lactobacillus in the composite lactobacillus fermentum is 1×10 8 ～2×10 9 cfu/g。
The invention also provides a fermented tea beverage, which is obtained by fermenting the composite lactobacillus starter at a constant temperature of between 35 and 40 ℃.
In one embodiment of the invention, the fermentation time is 10 to 14 hours.
The invention also provides application of the composite lactobacillus starter in preparing fermented tea beverage.
[ advantageous effects ]
(1) The invention provides a compound lactobacillus starter for fermented tea, which is fully dissolved in purified water and then fermented at 37 ℃, so that the viable count of lactobacillus in the fermented tea beverage obtained by fermentation is 1 multiplied by 10 6 Proliferation of cfu/mL to over 2X 10 9 In addition, cfu/mL is not easy to inactivate due to the active protective agent contained in the compound lactobacillus starter, and lactobacillus in the fermented tea drink obtained by fermenting the compound lactobacillus starter can have a certain active retention period under the refrigeration condition of 4 ℃.
(2) The invention provides a fermented tea beverage, which is obtained by fermenting composite lactobacillus starter at constant temperature of 35-40 ℃ for 10-14 h, wherein the viable count of lactobacillus in the fermented tea beverage can be up to 2 x 10 9 The cfu/mL or more has good beneficial effects, and in addition, as the composite lactobacillus starter used in the fermented tea beverage contains the active protective agent, the lactobacillus in the fermented tea beverage is not easy to inactivate, and the fermented tea beverage can have a certain active retention period under the refrigeration condition of 4 ℃.
Detailed Description
The invention is further illustrated below in conjunction with specific examples.
Lactobacillus plantarum CCFM8610 related to the following examples is described in the patent application text with the publication number of CN102827796A, and the preservation number is CGMCC No.6077; lactobacillus casei CCFM1074 as referred to in the following examples is described in the patent application publication No. CN110591986a with deposit number GDMCC No.60766; lactobacillus rhamnosus CCFM0528 related to the following examples is described in the patent application text with publication number CN103275905a, with preservation number CGMCC No.7317; lactobacillus reuteri CCFM1072 referred to in the following examples is described in the patent application publication No. CN110643542a with deposit number GDMCC No.60735; the yeast extracts, yeast extract, yeast protein, yeast peptone, soybean peptone, tryptone, casein peptone, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, potassium phosphate, sodium citrate, whey protein isolate, casein, soybean protein, collagen, skim milk, polysaccharide, dietary fiber, resistant dextrin, maltodextrin, polydextrose, oligosaccharide, hydroxyethyl starch, which are available from Shanghai Inset technology Co.
The following examples relate to the following media:
MRS solid medium: 10g/L peptone, 10g/L beef extract, 20g/L glucose, 2g/L sodium acetate, 5g/L yeast powder, 2g/L, K diammonium hydrogen citrate 2 HPO 4 ·3H 2 O 2.6g/L、MgSO 4 ·7H 2 O 0.1g/L、MnSO 4 ·H 2 O0.05 g/L, tween 801mL/L, agar powder 15g/L, and cysteine hydrochloride 0.5g/L.
MRS liquid medium: 10g/L peptone, 10g/L beef extract, 20g/L glucose, 2g/L sodium acetate, 5g/L yeast powder, 2g/L, K diammonium hydrogen citrate 2 HPO 4 ·3H 2 O 2.6g/L、MgSO 4 ·7H 2 O 0.1g/L、MnSO 4 ·H 2 O0.05 g/L, tween 801mL/L, cysteine hydrochloride 0.5g/L.
The preparation method of the lactobacillus powder in the following examples comprises the following steps:
drawing lines on an MRS solid culture medium by dipping lactobacillus liquid in a fungus-retaining tube with an inoculating loop, and culturing at a constant temperature of 37 ℃ for 36 hours to obtain single colonies; single colony is selected and inoculated in MRS liquid culture medium, and the culture is carried out for 12 hours at the constant temperature of 37 ℃ to obtain seed liquid; inoculating the seed solution into MRS liquid culture medium according to the inoculum size of 2% (v/v), and culturing at 37 ℃ for 24 hours to obtain culture solution; centrifuging the culture solution, and collecting thalli; suspending the strain in stachyose solution with the concentration of 200g/L to obtain heavy suspension; lyophilizing the resuspension to obtain lactobacillus powder;
wherein, the freeze-drying process comprises the following steps: placing the heavy suspension in an environment of-50 ℃ for 4 hours; primary drying at-30deg.C and 200 μbar, and maintaining for 30 hr; and (3) secondary drying, 25 ℃ and 0 μbar, and maintaining for 20h.
The detection method involved in the following examples is as follows:
measurement of viable count of lactic acid bacteria:
the national standard GB 4789.35-2016 food safety national standard food microbiology detection of lactobacillus detection is adopted.
Determination of lactic acid bacteria survival rate:
the survival rate of lactobacillus in different storage periods of the fermented tea soup is calculated according to the following formula:
survival rate= (number of viable bacteria of lactobacillus after storage of fermented tea beverage for different time/number of viable bacteria of lactobacillus when fermented tea beverage is stored for 0 day) ×100%.
Example 1: preparation of composite lactobacillus starter
The method comprises the following specific steps:
scheme one: crushing black tea leaves to 80 meshes to obtain crushed tea leaves; mixing the crushed tea with water, boiling and leaching for 30min to obtain tea soup, wherein the mass of the crushed tea accounts for 20 per mill of the mass of the water during leaching; centrifuging 3000g of tea soup for 10min to obtain supernatant; adding glucose, 5%o yeast protein, 2%o soybean peptone, 5%o dipotassium hydrogen phosphate, 20%o sodium citrate and 4% whey protein isolate accounting for 7% of the total mass of the supernatant into the supernatant to obtain a mixed solution; heating the mixed solution at 85deg.C for 15min for sterilization, vacuum concentrating at 45deg.C, and freeze drying to obtain tea powder; mixing tea powder and lactobacillus rhamnosus CCFM0528 powder in aseptic environment to obtain composite lactobacillus starter 1, wherein the viable count of lactobacillus rhamnosus CCFM0528 in the composite lactobacillus starter 1 is 1×10 9 cfu/g。
Scheme II: based on the first scheme, 7% of glucose is removed, or 7% of glucose is replaced by 6% of fructose, 6% of sucrose, 5% of lactose, 5% of maltose or 8% of fruit and vegetable juice respectively, so that 2-7 of the composite lactobacillus starter is obtained.
Scheme III: on the basis of the first scheme, 5 permillage of yeast protein and 2 permillage of soybean peptone are removed, or 5 permillage of yeast protein and 2 permillage of soybean peptone are respectively replaced by 8 permillage of yeast extract, 8 permillage of yeast extract powder, 5 permillage of yeast protein and 2 permillage of tryptone or 5 permillage of yeast peptone and 2 permillage of casein peptone, so that the composite lactobacillus starter 8-12 is obtained.
Scheme IV: on the basis of the scheme one, 5 permillage of dipotassium hydrogen phosphate and 20 permillage of sodium citrate are removed, or 5 permillage of dipotassium hydrogen phosphate and 20 permillage of sodium citrate are respectively replaced by 8 permillage of disodium hydrogen phosphate, 8 permillage of potassium dihydrogen phosphate and 15 permillage of sodium citrate, 5 permillage of potassium phosphate and 15 permillage of sodium citrate or 5 permillage of potassium sodium phosphate and 15 permillage of potassium citrate, so as to obtain the composite lactobacillus starter 13-17.
Scheme five: based on the first scheme, 4% of whey protein isolate is removed, or 4% of whey protein isolate is replaced by 4% of soybean protein, 3% of polysaccharide, 3% of maltodextrin or 4% of oligosaccharide respectively, so as to obtain the composite lactobacillus starter 18-22.
Scheme six: based on the first scheme, lactobacillus rhamnosus CCFM0528 is replaced by lactobacillus plantarum CCFM8610, lactobacillus casei CCFM1074 or lactobacillus reuteri CCFM1072 respectively to obtain the composite lactobacillus starter 23-25.
Example 2: preparation of fermented tea beverage
The method comprises the following specific steps:
scheme one: 3g of the composite lactobacillus fermentum 1 obtained in example 1 was dissolved in 500mL of purified water and fermented at 37℃for 12 hours to obtain a fermented tea beverage 1.
Scheme II: 3g of the composite lactobacillus leavening agent 2-7 obtained in the example 1 is respectively taken and dissolved in 500mL of purified water, and then fermented for 12 hours at 37 ℃ to obtain fermented tea beverages 2-7.
Scheme III: 3g of the composite lactobacillus starter obtained in example 1 was dissolved in 500mL of purified water, and then fermented at 37℃for 12 hours to obtain fermented tea beverages 8 to 12.
Scheme IV: 3g of the composite lactobacillus leavening agent 13-17 obtained in the example 1 is respectively taken and dissolved in 500mL of purified water, and then fermented for 12 hours at 37 ℃ to obtain the fermented tea beverage 13-17.
Scheme five: 3g of the composite lactobacillus leavening agent 18-22 obtained in the example 1 is respectively taken and dissolved in 500mL of purified water, and then fermented for 12 hours at 37 ℃ to obtain the fermented tea beverage 18-22.
Scheme six: 3g of the composite lactobacillus leavening agent 23-25 obtained in the example 1 is respectively taken and dissolved in 500mL of purified water, and then fermented for 12 hours at 37 ℃ to obtain the fermented tea beverage 23-25.
After the fermented tea beverage 1 to 25 was left at 4℃for 0, 6, 12 and 18 hours, the viable count of lactic acid bacteria in the fermented tea beverage 1 to 25 was measured (the measurement results are shown in Table 1).
As can be seen from Table 1, the method obtained in example 1 was usedWhen the composite lactobacillus starter 1 is used, the viable count of lactobacillus in the fermented tea beverage is highest and reaches 3.6X10 9 cfu/mL。
TABLE 1 viable count of lactic acid bacteria in fermented tea beverages for various times
Example 3: preparation of fermented tea beverage
The method comprises the following specific steps:
based on the first scheme of the embodiment 2, the black tea in the composite lactobacillus leavening agent 1 is replaced by jasmine tea, puer tea, tieguanyin or Dahongpao tea respectively to obtain fermented tea beverages 1-5.
The viable count of the lactic acid bacteria in the fermented tea beverage 1-5 is detected, and the detection result is as follows: the viable count of the lactic acid bacteria in the fermented tea beverage 1-5 is 3.6X10 respectively 9 cfu/mL、3.3×10 9 cfu/mL、5.4×10 7 cfu/mL、7.6×10 7 cfu/mL and 4.1X10 7 cfu/mL。
It can be seen that only black tea and jasmine tea can be used to prepare a fermented tea beverage rich in lactobacillus.
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (3)
1. The composite lactobacillus starter is characterized by comprising tea powder and lactobacillus powder; the tea powder is prepared by leaching tea leaves with water to obtain tea soup, adding soluble carbohydrate, multiplication factors, inorganic salt and an active protective agent into the tea soup to obtain mixed liquor, and finally drying the mixed liquor; the mass of the tea leaves accounts for 10-30 per mill of the mass of the water; the addition amount of the soluble carbohydrate in the tea soup accounts for 1-8% of the total mass of the tea soup; the addition amount of the proliferation factors in the tea soup is 2-10 per mill of the total mass of the tea soup; when the inorganic salt is one or more of dipotassium hydrogen phosphate, disodium hydrogen phosphate, monopotassium phosphate, sodium dihydrogen phosphate, potassium phosphate or sodium potassium phosphate, the adding amount of the inorganic salt in the tea soup accounts for 2-8 per mill of the total mass of the tea soup; when the inorganic salt is one or more than one of sodium citrate and potassium citrate, the adding amount of the inorganic salt in the tea soup is 9-20 per mill of the total mass of the tea soup; the tea is black tea and jasmine tea; the soluble carbohydrate is one or more of glucose, fructose, sucrose, lactose, maltose or fruit and vegetable juice; the proliferation factor is one or more than one of yeast extract, yeast extract powder, yeast protein, yeast peptone, soybean peptone, tryptone or casein peptone; the active protective agent is one or more of whey protein isolate, soybean protein and maltodextrin.
2. A fermented tea beverage, which is obtained by fermenting the compound lactobacillus starter according to claim 1 at a constant temperature of 35-40 ℃.
3. Use of the compound lactobacillus leavening agent of claim 1 for preparing a fermented tea beverage.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101748082A (en) * | 2008-12-11 | 2010-06-23 | 吉林省农业科学院 | Lactobacillus leavening agent, preparation method thereof and special bacterial strain |
| CN109527145A (en) * | 2017-09-22 | 2019-03-29 | 勐海茶业有限责任公司 | A kind of apple Pu'er tea mixed fermentation beverage and preparation method thereof |
| CN109864233A (en) * | 2018-09-14 | 2019-06-11 | 浙江李子园食品股份有限公司 | A kind of fermented apple juice and preparation method thereof rich in active plant lactobacillus |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101748082A (en) * | 2008-12-11 | 2010-06-23 | 吉林省农业科学院 | Lactobacillus leavening agent, preparation method thereof and special bacterial strain |
| CN109527145A (en) * | 2017-09-22 | 2019-03-29 | 勐海茶业有限责任公司 | A kind of apple Pu'er tea mixed fermentation beverage and preparation method thereof |
| CN109864233A (en) * | 2018-09-14 | 2019-06-11 | 浙江李子园食品股份有限公司 | A kind of fermented apple juice and preparation method thereof rich in active plant lactobacillus |
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