CN112450346B - Preparation method of sugar-free beer fermentation juice - Google Patents

Abstract

A preparation method of sugar-free beer fruit fermentation juice comprises the steps of raw material treatment, first yeast fermentation and second lactobacillus casei fermentation treatment, so that the sugar content of beer fruit juice is reduced from 13.7g/100mL to 0.119g/100mL, the national sugar-free food standard is achieved, and the requirements of diabetic consumer groups on low-sugar or sugar-free beverages can be met.

Description

Preparation method of sugar-free beer fermentation juice

Technical Field

The invention relates to the technical field of fruit juice foods and processing thereof, in particular to sugar-free fermented juice of Piteguo.

Technical Field

Pitot fruit original name, i.e. Pithecellobium praecox (local people also called "bud bread", sour pear and Pithecellobium praecox), is a unique ancient tree species growing on Taizi foot, has a planting history of more than 1000 years, is a local cultivar of Xinjiang pear series in apple subfamily of Rosaceae, is at the upper limit of pear plant distribution, and has the characteristics of strong tree vigor, long tree age, low soil requirement, damp-liking, cold resistance, disease and pest resistance, strong adaptability and the like. The Piteguo fruit has sweet and sour taste and warm nature, contains abundant proteins, reducing sugar, crude fibers, vitamins (C, B, B2), tannins, amino acids, potassium, calcium, iron and other mineral elements necessary for human bodies, and is a good food therapy product for moistening lung and nourishing stomach, relieving diarrhea and quenching thirst, softening blood vessels, dispelling alcohol and protecting liver. At present, the main way of processing the Piteguo is to prepare Piteguo juice, and researches are mainly focused on aspects of Piteguo juice beverage formulas, nutritional ingredients and the like. For example, yang Xuxing, ma Shu et al have formulated a beer juice beverage by studying the ratio of raw juice to sweetened sugar as follows: 30% of Piteguo juice, 5% of white granulated sugar, 0.1% of sodium cyclamate, 0.8% of caramel pigment, 0.8% of honey and 0.12% of citric acid. Ji Yong, luo Yan the best formulation for a juice beverage of Piteguo fruit determined from Piteguo fruit as a raw material is: 30% of Piteguo juice, 6% of white granulated sugar, 0.8% of mountain flower honey, 0.10% of caramel pigment and 0.06% of citric acid. However, the beer fruit beverage prepared by the existing formula has higher sugar content and is not suitable for diabetics.

The national GB 25080 standard prescribes that the low-sugar food is less than or equal to 5 g/100mL (liquid), sugar-free or sugar-free is less than or equal to 0.5 g/100mL (liquid), and the demand for low-sugar juice is on the rising trend due to the increasing year by year of diabetics in China. The most effective and simple method for reducing the sugar content of the juice is to use microorganism fermentation to consume sugar substances in the juice. Therefore, development of low-sugar beer fermentation juice, which meets the consumer group demand of diabetics, is urgent.

Disclosure of Invention

The invention provides a preparation method of sugar-free beer fermentation juice, which can meet the demands of diabetics consumer groups on low-sugar or sugar-free beverages.

The technical scheme adopted by the invention is as follows:

a preparation method of sugar-free beer fruit fermentation juice comprises the following specific steps:

(1) And (3) raw material treatment: sterilizing the Piteguo juice with 8 ° Brix of soluble solids content at 121deg.C for 20min, and cooling to room temperature for use;

(2) Primary fermentation: inoculating a compound bacteria liquid with the mass ratio of Angel red wine yeast to Hansen yeast of the family being 1:1, wherein the dosage is 1.0% -2.0% of the mass of fruit juice, and performing aerobic fermentation for 36-44 h at 26-30 ℃;

(3) And (3) secondary fermentation: after the fermentation of the saccharomycetes is finished, inoculating lactobacillus casei suspension, wherein the dosage of the lactobacillus casei suspension is 4-6% of the mass of the fruit juice, performing aerobic fermentation for 11-21h at the temperature of 35-39 ℃, and then heating the fermentation liquor to the temperature of 60-65 ℃ and keeping the temperature for 25-30min for secondary sterilization;

(4) And (3) packaging: and filling the fermentation raw juice into an aluminum-plastic composite bag by adopting a sterile filling machine and sealing.

Advantageous effects or advantages of the invention

(1) 2 high-sugar-resistance yeasts and lactobacillus casei are selected, and the sugar content of the Piteguo juice is reduced from 13.7g/100mL to 0.119g/100mL by optimizing the yeast compounding proportion and the fermentation process conditions of the yeasts and the lactobacillus casei, so that the national sugar-free food standard is achieved.

(2) The selected probiotics lactobacillus casei can effectively reduce the alcohol content while fermenting and reducing the blood sugar, and improve the flavor and quality of the product.

(3) The sugar-free probiotic-containing beer juice produced by adopting the two-stage fermentation is beneficial to health and has wide application.

Drawings

FIG. 1 is a graph of a single-factor test (inoculum size) of a composite yeast of the invention;

FIG. 2 is a graph of a single factor test of a composite yeast of the present invention (fermentation time);

FIG. 3 is a graph of a single factor test of a composite yeast of the present invention (fermentation temperature);

FIG. 4 is a graph of a single factor test of Lactobacillus casei of the present invention (inoculum size);

FIG. 5 is a graph of single factor test of Lactobacillus casei (fermentation time) according to the present invention;

FIG. 6 is a graph of a single factor test of Lactobacillus casei (fermentation temperature) according to the present invention.

Detailed Description

The invention is further illustrated by the following detailed description and the principles of the formulation.

The principle of selecting strains in the secondary fermentation process is as follows:

yeast is a single-cell fungus that can survive both aerobically and anaerobically, and is a natural starter. The EMP pathway is the main glycolytic pathway of yeasts and is also a common pathway for glucose to undergo aerobic or anaerobic metabolism. Saccharomycetes can decompose sugar into carbon dioxide and water under the condition of oxygen; under anaerobic conditions, yeasts decompose sugar into alcohol and carbon dioxide. Since the saccharomycetes are numerous, and the sugar content of the Pitot juice is high (12-14%), the selection of the high sugar-resistant saccharomycetes for fermentation is important.

Lactic acid bacteria are important strains of traditional lactic acid fermented foods, and have the potential effect of improving body health. The main metabolites in the anaerobic fermentation process of lactic acid bacteria are acids and enzymes. After the fermentation of the saccharomycetes is finished, lactobacillus is inoculated for fermentation, ethanol and the produced carboxylic acid can react to generate ester substances, and partial ethanol generated by the fermentation of the saccharomycetes is neutralized. Meanwhile, the produced lactic acid, extracellular polysaccharide and generated ethyl acetate can increase the flavor and taste of the product and are beneficial to health.

The Angel red wine yeast (Hubei Angel active dry Yeast Co., ltd.) and the Hansen pure Saccharomyces cerevisiae (chr. Hansen Holding A/S) adopted by the invention have the characteristics of high glucose resistance, acid resistance, high fermentation speed and the like. The lactobacillus casei (China general microbiological culture Collection center, strain number: 1.8727) belongs to probiotics, is suitable for low pH value conditions, can ferment fructose, galactose and glucose, cannot utilize melibiose, raffinose and xylose, cannot decompose arginine to produce ammonia, and the generated organic acid is mainly lactic acid, acetic acid, phenyllactic acid, propionic acid, malic acid and citric acid. Lactobacillus casei has good acid resistance and bile resistance, and can reduce plasma cholesterol, enhance host nonspecific resistance to microbial pathogens, promote removal of pathogens in intestinal tract, treat intestinal flora disorder and enhance intestinal permeability, thereby preventing food allergy and acute diarrhea. In addition, lactobacillus casei can increase anti-low density oxidized lipid antibody and lymphocyte, obviously enhance granulocyte phagocytosis, and perform immunoregulation on host to prevent tumor. Lactobacillus casei is widely used as a probiotic for functional foods due to its ability to survive stably in the human intestinal tract and its numerous medical and health functions.

The juice produced by combining sugar-free and probiotics has wide application and market prospect, and if producing oral liquid or beverage, the juice is characterized in that: xylitol as a functional sweetener can be selected to avoid the rise of blood sugar level; the probiotics with unique health care function can help digestion, prevent constipation and cell aging, resist tumor and regulate human body functions. Thus, at present, the development of sugar-free products and probiotic products will be a future development direction, as food products are being developed towards natural and functional types.

Based on the principle, the specific implementation mode of the invention is as follows:

example 1, a method for preparing sugar-free beer fermentation juice, which comprises the following steps:

(1) 100kg of Piteguo juice with the content of soluble solids of 8 DEG Brix qualified by inspection is sterilized at 121 ℃ for 20min and cooled to room temperature.

(2) Primary fermentation: inoculating Angel red wine yeast (Hubei Angel active dry Yeast Co., ltd., china) and Hansen Yeast (Chur. Hansen Holding A/S) at a mass ratio of 1:1, fermenting with oxygen at 28deg.C for 40 hr.

(3) And (3) secondary fermentation: after the fermentation of the saccharomycetes is finished, lactobacillus casei suspension is inoculated, the dosage is 5 percent of the mass of the fruit juice, the fermentation is carried out for 16 hours at 37 ℃, and then the fermentation liquor is heated to 65 ℃ and kept for 25 minutes for secondary sterilization. Lactobacillus casei was purchased from the China general microbiological culture Collection center, strain number: 1.8727.

(4) And (3) packaging: filling the fermentation raw juice into an aluminum-plastic composite bag by adopting a sterile filling machine, sealing and obtaining the specification

20kg。

The final sugar content of this example was measured to be 0.119/g/100 mL by secondary fermentation.

Example 2, a method for preparing sugar-free beer fermentation juice, comprises the following specific implementation steps:

(1) 100kg of Piteguo juice with the content of soluble solids of 8 DEG Brix qualified by inspection is sterilized at 121 ℃ for 20min and cooled to room temperature.

(2) Primary fermentation: inoculating Angel red wine yeast (Hubei Angel active dry Yeast Co., ltd., china) and Hansen Yeast (Chur. Hansen Holding A/S) at a mass ratio of 1:1, fermenting with oxygen at 30deg.C for 36 hr.

(3) And (3) secondary fermentation: after the fermentation of the saccharomycetes is finished, lactobacillus casei suspension is inoculated, the dosage is 5 percent of the mass of the fruit juice, the fermentation is carried out for 21 hours at 35 ℃, and then the fermentation liquor is heated to 65 ℃ and kept for 25 minutes for secondary sterilization. Lactobacillus casei was purchased from the China general microbiological culture Collection center, strain number: 1.8727.

(4) And (3) packaging: filling the fermentation raw juice into an aluminum-plastic composite bag by adopting a sterile filling machine, sealing and obtaining the specification

20kg。

The final sugar content of the sample obtained in this example was 0.1551g/100mL by secondary fermentation.

Example 3, a method for preparing sugar-free beer fermentation juice, comprises the following specific implementation steps:

(1) 100kg of Piteguo juice with the content of soluble solids of 8 DEG Brix qualified by inspection is sterilized at 121 ℃ for 20min and cooled to room temperature.

(2) Primary fermentation: inoculating Angel red wine yeast (Hubei Angel active dry Yeast Co., ltd., china) and Hansen Yeast (chr. Hansen Holding A/S) at a mass ratio of 1:1, fermenting with 2% of the fruit juice mass under aerobic condition at 26deg.C for 40 hr.

(3) And (3) secondary fermentation: after the fermentation of the saccharomycetes is finished, lactobacillus casei suspension is inoculated, the quality of the juice is 6%, the juice is subjected to aerobic fermentation for 11 hours at 37 ℃, and then the fermentation liquor is heated to 63 ℃ and kept for 28min for secondary sterilization. Lactobacillus casei was purchased from the China general microbiological culture Collection center, strain number: 1.8727.

(4) And (3) packaging: filling the fermentation raw juice into an aluminum-plastic composite bag by adopting a sterile filling machine, sealing and obtaining the specification

20kg。

The final detection of sugar content of this example was 0.1698 g/100mL by secondary fermentation.

Example 4, a method for preparing sugar-free beer fermentation juice, the specific implementation method is as follows:

(1) 100kg of Piteguo juice with the content of soluble solids of 8 DEG Brix qualified by inspection is sterilized at 121 ℃ for 20min and cooled to room temperature.

(2) Primary fermentation: inoculating Angel red wine yeast (Hubei Angel active dry Yeast Co., ltd., china) and Hansen Yeast (Chur. Hansen Holding A/S) at a mass ratio of 1:1, fermenting with 1% of fruit juice mass under aerobic condition at 28deg.C for 44 hr.

(3) And (3) secondary fermentation: after the fermentation of the saccharomycetes is finished, inoculating lactobacillus casei suspension, using 4% of the mass of the fruit juice, performing aerobic fermentation for 16 hours at 39 ℃, and then heating the fermentation liquor to 60 ℃ and keeping for 30min for secondary sterilization. Lactobacillus casei was purchased from the China general microbiological culture Collection center, strain number: 1.8727.

(4) And (3) packaging: filling the fermentation raw juice into an aluminum-plastic composite bag by adopting a sterile filling machine, sealing and obtaining the specification

20kg。

The final detection of sugar content of this example was 0.1602 g/100mL by secondary fermentation.

The invention is further illustrated below in terms of recipe and process parameter determination.

(1) And (3) raw material treatment: diluting or concentrating the juice to soluble solid content of 8 ° Brix, sterilizing at 121deg.C for 20min, and cooling to room temperature.

(2) Determining the proportion of saccharomycetes strains: the method comprises the steps of selecting Angel red wine yeast (Hubei Angel active dry Yeast Co., ltd., china) and Hansen yeast (Chr. Hansen Holding A/S) with different mass ratios, fermenting at 28 ℃ for 40h, inoculating 5% lactobacillus casei, fermenting at 37 ℃ for 16h, and measuring the soluble solid content of a sample.

(3) Process optimization test

1) Single factor test design

A. Composite saccharomycetes: the method (Wang Yu. Study on preparation of low-sugar red date juice based on composite fermentation method [ D ]. Hebei university of agriculture, 2015.) is slightly modified, and under the basic conditions that the mass ratio of Angel red wine yeast to Hansen yeast is 1:1, the inoculation amount of yeast is 1.5% of the mass of fruit juice, the fermentation time of yeast is 40h, and the fermentation temperature is 28 ℃, other conditions are unchanged, only one factor is changed to perform a single factor test in the fermentation stage of yeast, the content of soluble solids is used as an index, and the optimal parameters are optimized. Wherein, the parameters of the inoculation amount of the saccharomycetes are set to 0.5%, 1%, 1.5%, 2% and 2.5%, the parameters of the fermentation temperature of the saccharomycetes are set to 26 ℃, 28 ℃, 30 ℃, 32 ℃ and 34 ℃ and the parameters of the fermentation time of the saccharomycetes are set to 28h, 32h, 36h, 40h and 44h.

B. Lactobacillus casei: reference is made to the method Liu Renlu (Liu Renlu. Process study of fermentation of wort by composite strains [ D ]. University of light industry of Wuhan, 2015.) with minor modifications. Inoculating a compound bacteria liquid with the mass ratio of Angel red wine yeast to Hansen yeast of Hansen family of 1:1, inoculating yeast with the mass of fruit juice of 1.5%, fermenting at 28deg.C for 40h, and inoculating a bacteria suspension. The method is characterized in that the content of soluble solids is used as a research index, the inoculation amount of lactobacillus casei is fixed to be 5% of the quality of fruit juice, the fermentation time is 16h, the fermentation temperature is 37 ℃, and the influence of the inoculation amount of lactobacillus, 3%, 4%, 5% and 6% on the content of the soluble solids of the beer juice, the fermentation time of 6h, 11h, 16h, 21h, 26h and the fermentation temperature of 31 ℃, 33 ℃, 35 ℃, 37 ℃ and 39 ℃ is respectively researched under the basic condition that the fermentation temperature is 37 ℃.

2) Response surface optimization test design

Based on a single factor test, design-expert8.0 software is adopted to carry out Box-Behnken test Design. Three factors of inoculum size (A), fermentation time (B) and fermentation temperature (C) are selected, and three-factor three-level test design is carried out by taking total sugar content (mg/mL) as a response value. The yeast factor levels are shown in Table 1 and the Lactobacillus casei factor levels are shown in Table 2.

TABLE 1 Yeast response surface test factor level and coding

TABLE 2 Lactobacillus casei response surface test factor level and coding

3) Determination of total sugar content: after the fermentation of yeasts, reference is made to the method of Lv Wei (Lv Wei. Research on the preparation of fructooligosaccharides from garlic [ D ]. Nanchang: nanchang university, 2010.) the total sugar content in the fermentation broth is determined by the phenol sulfuric acid method.

(4) Test results

1) Single factor test results

In the range of 0.5% -1.5% of yeast inoculum size, the soluble solids content of the Piteguo juice is reduced with the increase of inoculum size, and the soluble solids content is the lowest when the inoculum size is 1.5%. As the inoculum size increased (2% -2.5%), the soluble solids content increased, but not so much (FIG. 1). The range of inoculum size optimized for this subsequent trial was therefore chosen to be between 1% and 2%. In the range of 28h-44h of saccharomycete fermentation time, the content of the soluble solid matters is in a trend of decreasing and then increasing along with the prolongation of the fermentation time, and the content of the soluble solid matters reaches the minimum value when the fermentation time is 40h (figure 2), so that the fermentation time range optimized by the subsequent experiment is selected to be between 36h and 44h. The soluble solid content of the yeast is reduced to different degrees along with the rise of the fermentation temperature within the range of 26-34 ℃, and the sugar reducing effect is best (figure 3) at the fermentation temperature of 28 ℃, and the lowest value of the yeast is taken as the middle value to be taken into the two ends in consideration of shortening the fermentation time and reducing the cost. Thus, the fermentation temperature range optimized for the subsequent experiments was selected between 26℃and 30 ℃.

In the range of 2% -6% of lactobacillus casei inoculum size, the soluble solids content of the raw juice of the Piteguo is reduced with the increase of inoculum size, and at 5% of inoculum size, the soluble solids content is the lowest, and with the increase of inoculum size (6%), the soluble solids content is increased (FIG. 4). Thus, 5% was chosen for subsequent experimental optimization. In the range of 6-26 h of lactobacillus casei fermentation time, the content of soluble solids is reduced and then increased along with the prolongation of the fermentation time, and the content of the soluble solids reaches the minimum value at 16h of the fermentation time (figure 5), so that 16h is selected for subsequent experimental optimization. The soluble solids content tended to decrease and then increase with increasing fermentation temperature in the range of 31℃to 39℃for Lactobacillus casei, and was the lowest at 37 ℃. Thus, 37℃was chosen for subsequent experimental optimisation.

In the process of process optimization, the primary fermentation, namely the saccharomycete fermentation stage, is the main fermentation stage, the sugar substances are basically consumed, and the secondary fermentation stage, namely the lactobacillus fermentation stage, is used for enabling acid substances generated by the lactobacillus casei probiotics during fermentation to be combined with ethanol generated by the saccharomycete fermentation to generate ester substances, so that the flavor of the Piteger fruits is improved, and the quality of the Piteger fruits is improved.

2) Response surface test results

TABLE 3 Yeast response surface test results

TABLE 4 regression equation analysis of variance

Note that: * Represent a significant level difference of 0.01; * Indicating a significant level difference of 0.05

Performing quadratic polynomial regression fitting on the table 4 data by using Design-Expert8.0 software to obtain a fitting equation: total sugar = 199.42778-10.88585A-1.19575B-11.698012 +0.10562ab-0.083500 AC-0.0127719 bc +2.88420a ² +0.017769B ² +0.21764C ²

Analysis of variance (Table 4) of regression model, model P<0.01, indicating that the model equation is highly significant, the quench test p= 0.2538>0.05, the model mismatch term is not obvious, and the model is properly selected. Correlation coefficient R ² =0.9954，R _Adj ² The equation is better fit to the test, and can also better reflect the relation between each factor and the response value, and the optimal formula of the sugar-free Piteger fermentation juice is possible to be predicted by using the equation model analysis.

TABLE 5 Lactobacillus casei response surface test results

TABLE 6 regression equation analysis of variance

Note that: * Represent a significant level difference of 0.01; * Indicating a significant level difference of 0.05

Performing quadratic polynomial regression fitting on the table 6 data by using Design-Expert8.0 software to obtain a fitting equation: total sugar = 147.62972-5.05937a-0.55689B-6.99231C-0.015750ab+0.045625ac+2.6e-003bc+0.35255A ² +0.016252B ² +0.090950C ²

Analysis of variance (Table 6) of regression model, model P<0.01, indicating that the model equation is highly significant, the quench test p= 0.2359>0.05, the model mismatch term is not obvious, and the model is properly selected. Correlation coefficient R ² =0.9886，R _Adj ² The equation is better fit to the test, and can also better reflect the relation between each factor and the response value, and the optimal formula of the sugar-free Piteger fermentation juice is possible to be predicted by using the equation model analysis.

3) Optimal recipe determination

The optimal technological parameters of the fermentation of the sugar-free Piteguo juice are predicted by a model: the inoculation amount of the saccharomycetes is 1.5%, the fermentation time is 40h, the fermentation temperature is 28 ℃, the inoculation amount of the lactobacillus casei is 5%, the fermentation time is 16h, and the fermentation temperature is 37 ℃. And verifying the validity of the regression model according to the obtained optimal parameters. 3 verification tests show that the average total sugar content of the beer juice after fermentation is 0.119g/100mL, and the actual measured value is 0.0326 higher than the theoretical predicted value, but the two values are close and have no obvious difference. The model is reasonable and effective, and the regression equation can well predict the theoretical total sugar content and has certain practical guiding significance.

Claims (1)

1. A preparation method of sugar-free beer fermentation juice is characterized by comprising the following specific steps:

(1) And (3) raw material treatment: sterilizing the Piteguo juice with 8 ° Brix of soluble solids content at 121deg.C for 20min, and cooling to room temperature for use;

(2) Primary fermentation: inoculating compound bacteria liquid of Angel red wine yeast and Hansen yeast with mass ratio of 1:1, and performing aerobic fermentation at 28deg.C for 40 hr;

(3) And (3) secondary fermentation: after the fermentation of the saccharomycetes is finished, inoculating lactobacillus casei suspension, wherein the dosage is 5% of the mass of the fruit juice, performing aerobic fermentation for 16 hours at 37 ℃, and then heating the fermentation liquor to 60-65 ℃ and keeping for 25-30min for secondary sterilization;

(4) And (3) packaging: and filling the fermentation raw juice into an aluminum-plastic composite bag by adopting a sterile filling machine and sealing.

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