WO2018103118A1 - 一种发酵型大豆乳清饮料及其制备方法 - Google Patents

一种发酵型大豆乳清饮料及其制备方法 Download PDF

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WO2018103118A1
WO2018103118A1 PCT/CN2016/109536 CN2016109536W WO2018103118A1 WO 2018103118 A1 WO2018103118 A1 WO 2018103118A1 CN 2016109536 W CN2016109536 W CN 2016109536W WO 2018103118 A1 WO2018103118 A1 WO 2018103118A1
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lactic acid
acid bacteria
soybean whey
fermentation
fermented
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何鑫平
梁岩
于配配
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Shenzhen Institute of Advanced Technology of CAS
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Shenzhen Institute of Advanced Technology of CAS
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF
    • A23C11/00—Milk substitutes, e.g. coffee whitener compositions
    • A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
    • A23C11/06—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing non-milk proteins

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  • the invention relates to the technical field of food processing methods, in particular to a fermented soybean whey beverage and a preparation method thereof.
  • Soy whey is a by-product of soy protein processing. It is rich in whey protein, soybean oligosaccharides, inorganic salts, etc. Currently, it is directly discharged by waste, causing waste of resources and causing environmental pollution. Soy oligosaccharides in soy whey are a kind of functional sugar. The main components are sucrose, stachyose and raffinose. Except for sucrose, the other is not absorbed by the body, and can directly reach the intestine and promote the intestine. Bifidobacteria proliferate, can inhibit harmful bacteria, enhance beneficial bacteria, laxative, remove toxins, and improve intestinal immune function. Sucrose is a common disaccharide. Excessive intake can cause obesity and tooth decay, which is also extremely detrimental to diabetic patients.
  • patent 201510899907.0 discloses a method for extracting oligosaccharides in soybean deep processing, which is super-pretreated by soybean milk. Filtration, nanofiltration, spray drying to obtain high-purity oligosaccharides, and increase the added value of the soybean processing industry
  • Patent 201210393907.X discloses a health beverage using yellow pulp water as a main raw material and a preparation method thereof, which is soybean yellow pulp Water is the main raw material, and sucrose, fruit juice, sour agent, food flavor, and bean dregs fermentation broth are added to make a health drink.
  • the soybean product by-products are fully utilized to solve the problem of treatment of yellow pulp water and bean dregs;
  • the prior art methods have many disadvantages, such as the patent 201510899907.0 purification method, the oligosaccharide is not high in purity, the process is cumbersome, and is not suitable for large-scale production, and the energy consumption is high, the cost is large, and some oligosaccharides are inevitably produced. Loss of ingredients; bioactive substances contained in products made according to the method of patent 201210393907.X, such as low content of soybean oligosaccharides, have limited effect.
  • a first object of the present invention is to provide a method for preparing a fermented soybean whey beverage, wherein the fermented soybean whey beverage preparation method can efficiently maintain the lactic acid bacteria catalysis in the fermentation liquid while efficiently utilizing soybean industrial by-products.
  • the activity can also recycle the immobilized lactic acid bacteria after the continuous fermentation reaction, and the process is simple and the cost is low; in addition, the sucrose in the soybean whey is effectively removed, and the soybean oligosaccharide active ingredients such as stachyose and raffinose are sufficiently retained.
  • a second object of the present invention is to provide a fermented soybean whey beverage prepared by the above-described method for preparing a fermented soybean whey beverage, wherein the fermented soybean whey beverage has a low sucrose content, stachyose, Soybean saccharide and other soybean oligosaccharide active ingredients are high in content, can effectively promote the proliferation of human intestinal bifidobacteria, increase beneficial bacteria, inhibit harmful bacteria, laxative, remove toxins, and improve human intestinal immune function.
  • the invention relates to a method for preparing a fermented soybean whey beverage, which comprises the soybean whey as a raw material, which is sequentially concentrated and compounded, and then mixed with the immobilized lactic acid bacteria colloidal particles to obtain a soybean whey beverage.
  • the preparation method of the fermented soybean whey beverage of the invention effectively overcomes the backwardness of the extraction technology of the active component such as soybean oligosaccharide in the existing soybean whey, and the activity of the soybean oligosaccharide in the obtained product is
  • the invention adopts immobilized lactic acid bacteria glue for fermentation, while efficiently utilizing soybean industrial by-products,
  • the lactic acid bacteria catalytic activity in the fermentation liquid can be maintained, and the immobilized lactic acid bacteria can be recovered after the continuous fermentation reaction, and the process is simple and the cost is low.
  • the concentration comprises concentrating the soy whey by heating under pressure to a concentration of soybean whey of 10% by weight to 15% by weight, preferably 12% by weight to 13% by weight, further preferably 13% by weight.
  • the heating temperature is 50 to 70 ° C, preferably 55 to 65 ° C, and more preferably 60 ° C.
  • the pressurizing pressure is 80 to 90 kPa, preferably 83 to 87 kPa, and further preferably 85 kPa.
  • the compounding comprises: compounding the concentrated soybean whey with milk powder, and sterilizing to obtain a fermentation starting material, wherein the milk powder is added in an amount of 5%-20%, preferably 10%, of the mass of the obtained fermentation starting material. -20%, further preferably 15%.
  • the preparation method of the immobilized lactic acid bacteria colloid comprises: adding sterile sodium alginate to a lactic acid bacteria suspension selectively using sucrose, mixing, adding the obtained mixture to a CaCl 2 solution, and standing still.
  • the bacteria are washed with physiological saline to obtain immobilized lactic acid bacteria colloidal particles having a diameter of 2-3 mm, preferably 2.5-3 mm, and more preferably 2.5 mm.
  • the sterile sodium alginate is added in an amount of from 2% to 2.5%, preferably from 2.3% to 2.5%, further preferably 2.3%, based on the mass of the lactic acid bacteria suspension.
  • the concentration of the CaCl 2 solution is from 2% to 4%, preferably from 3% to 4%, further preferably 3%.
  • the fermenting comprises: mixing the immobilized lactic acid bacteria colloidal particles with the raw material to be fermented, adding sodium acetate, thermostatically fermenting, filtering, removing the immobilized lactic acid bacteria colloidal particles, and sterilizing to obtain the soybean whey beverage.
  • the mass ratio of the immobilized lactic acid bacteria colloid to the raw material to be fermented is 1:3-4, preferably 1:3.5-4, further preferably 1:3.5.
  • the sodium acetate is added in an amount of 0.2% to 0.6%, preferably 0.3% to 0.5%, further preferably 0.4%, based on the mass of the raw material to be fermented.
  • the fermentation temperature of the thermostatic fermentation is 37-42 ° C, preferably 38-40 ° C, further preferably 39 ° C;
  • the fermentation time of the isothermal fermentation is 24-48 h, preferably 24-36 h, further preferably 36 h.
  • the immobilized lactic acid bacteria colloidal particles obtained after the filtration are washed with sterile physiological saline, stored in a low temperature physiological saline, and reused.
  • a fermented soybean whey beverage prepared by the above-described method for preparing a fermented soybean whey beverage.
  • the fermented soybean whey beverage of the invention has low sucrose content, high content of soybean oligosaccharide active ingredients such as stachyose and raffinose, can effectively promote the proliferation of human intestinal bifidobacteria, increase beneficial bacteria, inhibit harmful bacteria, and moisten
  • the bowels are laxative, remove toxins, and improve the intestinal immune function of the human body.
  • the preparation method of the fermented soybean whey beverage of the invention effectively overcomes the backward extraction technology of the active ingredient such as soybean oligosaccharide in the existing soybean whey, and the low content of the active ingredient such as soybean oligosaccharide in the obtained product, the soybean whey related product
  • the invention has the disadvantages or disadvantages of high energy consumption, high cost, unclear product efficacy, etc.
  • the invention adopts immobilized lactic acid bacteria gel for fermentation, and can effectively maintain the catalytic activity of lactic acid bacteria in the fermentation liquid while efficiently utilizing soybean industrial by-products.
  • the immobilized lactic acid bacteria can be recycled after the continuous fermentation reaction, and the process is simple and the cost is low.
  • the fermented soybean whey beverage of the invention has low sucrose content, high content of soybean oligosaccharide active ingredients such as stachyose and raffinose, can effectively promote the proliferation of human intestinal bifidobacteria, increase beneficial bacteria, inhibit harmful bacteria, and moisten
  • the bowels are laxative, remove toxins, and improve the intestinal immune function of the human body.
  • FIG. 1 is a process flow diagram of a method for preparing a fermented soybean whey beverage provided by a specific embodiment of the present invention.
  • FIG. 1 is a process flow diagram of a method for preparing a fermented soybean whey beverage provided by a specific embodiment of the present invention.
  • the invention provides a preparation method of a fermented soybean whey beverage, which comprises the soybean whey as a raw material, which is sequentially concentrated and compounded, and then mixed with the immobilized lactic acid bacteria colloidal particles to obtain a soybean whey beverage.
  • the preparation method of the fermented soybean whey beverage of the invention effectively overcomes the backward extraction technology of the active ingredient such as soybean oligosaccharide in the existing soybean whey, and the low content of the active ingredient such as soybean oligosaccharide in the obtained product, the soybean whey related product
  • the invention has the disadvantages or disadvantages of high energy consumption, high cost, unclear product efficacy, etc.
  • the invention adopts immobilized lactic acid bacteria gel for fermentation, and can effectively maintain the catalytic activity of lactic acid bacteria in the fermentation liquid while efficiently utilizing soybean industrial by-products.
  • the immobilized lactic acid bacteria can be recycled after the continuous fermentation reaction, and the process is simple and the cost is low.
  • the soybean whey may select a soybean whey by-product produced from a factory for producing soy protein isolate according to the difficulty of obtaining the raw material, or use the soybean meal as a raw material, and adopt the traditional alkali-soluble acid precipitation method to soybean Soybean whey is obtained by crushing, sieving, alkali-soluble, acid-sinking, and centrifugation.
  • the concentration comprises concentrating the soy whey by heating under pressure to a concentration of soybean whey of 10% by weight to 15% by weight, preferably 12% by weight to 13% by weight, further preferably 13% by weight.
  • Appropriate concentration of soy whey before fermentation helps to improve the efficiency of subsequent fermentation, promote the full metabolism of sucrose, and improve product quality.
  • the heating temperature is 50 to 70 ° C, preferably 55 to 65 ° C, and more preferably 60 ° C.
  • the pressurizing pressure is 80 to 90 kPa, preferably 83 to 87 kPa, and further preferably 85 kPa.
  • Evaporation and concentration of soy whey at a specific temperature and pressure helps to increase the concentration efficiency.
  • the evaporation concentration is carried out using a rotary evaporator.
  • the compounding comprises: compounding the concentrated soybean whey with milk powder, and sterilizing to obtain a fermentation starting material, wherein the milk powder is added in an amount of 5%-20%, preferably 10%, of the mass of the obtained fermentation starting material. -20%, further preferably 15%.
  • the concentrated soybean whey is compounded with the milk powder.
  • the milk powder also provides a part of the lactic acid bacteria metabolism raw material, and the addition of the milk powder can also improve the nutritional value of the obtained fermented product and improve the taste and taste.
  • the milk powder comprises milk powder and/or goat milk powder.
  • the milk powder comprises whole milk powder and/or skimmed milk powder, preferably comprising skimmed milk powder.
  • the preparation method of the immobilized lactic acid bacteria colloid comprises: adding sterile sodium alginate to a lactic acid bacteria suspension selectively using sucrose, mixing, adding the obtained mixture to a CaCl 2 solution, and standing still.
  • the bacteria are washed with physiological saline to obtain immobilized lactic acid bacteria colloidal particles having a diameter of 2-3 mm, preferably 2.5-3 mm, and more preferably 2.5 mm.
  • the invention adopts a specific method to prepare immobilized lactic acid bacteria colloidal particles with a specific particle size, and adopts the obtained immobilized lactic acid bacteria gel for fermentation, and can effectively maintain the catalytic activity of the lactic acid bacteria in the fermentation liquid while efficiently utilizing the soybean industrial by-products.
  • the immobilized lactic acid bacteria are recovered after the continuous fermentation reaction.
  • the sterile sodium alginate is added in an amount of from 2% to 2.5%, preferably from 2.3% to 2.5%, further preferably 2.3%, based on the mass of the lactic acid bacteria suspension.
  • Sterile sodium alginate can be used as an emulsion stabilizer and thickener.
  • the concentration of the CaCl 2 solution is from 2% to 4%, preferably from 3% to 4%, further preferably 3%.
  • calcium chloride acts as a sequestering agent and a curing agent.
  • the immobilized lactic acid bacteria colloidal particles having a specific particle size are obtained by using a specific amount of sterile sodium alginate and calcium chloride.
  • sterile sodium alginate is added to a suspension of lactic acid bacteria selectively utilizing sucrose, and after mixing, the resulting mixed droplets are added to the CaCl 2 solution.
  • the lactic acid bacteria suspension can be obtained by self-cultivation, screening and configuration.
  • the lactic acid bacteria suspension employed in the embodiment of the present invention is obtained by the following method:
  • the sucrose was replaced by glucose in the MRS medium to prepare a No. 1 MRS medium containing no glucose.
  • the sucrose and soybean oligosaccharide were mixed in a ratio of 1:1 to replace the glucose in the MRS medium to prepare a No. 2 MRS medium containing no glucose.
  • the lyophilized lactic acid bacteria strain was picked and inoculated on the No. 1 MRS agar plate medium containing no glucose. After anaerobic culture, a single colony was picked in 5 mL of glucose-free MRS liquid medium, anaerobic. After the culture, the amount of sucrose and soybean oligosaccharide residual sugar was measured.
  • the fermenting comprises: mixing the immobilized lactic acid bacteria colloidal particles with the raw material to be fermented (that is, the fermentation raw material mentioned above), adding sodium acetate, thermostatically fermenting, filtering, removing the immobilized lactic acid bacteria colloidal particles, and sterilizing the soybean milk. Clear drink.
  • the invention adopts immobilized lactic acid bacteria gel for fermentation, and can effectively utilize the soy industrial by-products while maintaining the catalytic activity of the lactic acid bacteria in the fermentation liquid, and recycling the immobilized lactic acid bacteria after the continuous fermentation reaction.
  • the mass ratio of the immobilized lactic acid bacteria colloid to the raw material to be fermented is 1:3-4, preferably 1:3.5-4, further preferably 1:3.5.
  • the immobilized lactic acid bacteria colloidal particles and the raw materials to be fermented in a specific amount ratio help to improve the fermentation efficiency, promote the full metabolism of sucrose, and improve the product quality.
  • the sodium acetate is added in an amount of 0.2% to 0.6%, preferably 0.3% to 0.5%, further preferably 0.4%, based on the mass of the raw material to be fermented.
  • sodium acetate in a specific amount can inhibit the growth of other bacteria and ensure the growth and proliferation of lactic acid bacteria.
  • the fermentation temperature of the isothermal fermentation is 37-42 ° C, preferably 38-40 ° C, and further preferably 39 ° C.
  • the fermentation time of the isothermal fermentation is 24-48 h, preferably 24-36 h, further preferably 36 h.
  • the use of specific fermentation temperature and fermentation time helps to improve fermentation efficiency, promote sucrose full metabolism, and improve product quality.
  • the immobilized lactic acid bacteria colloidal particles obtained after the filtration are washed with sterile physiological saline, stored in a low temperature physiological saline, and reused.
  • the low temperature is 10 ° C or lower, preferably 5 ° C or lower, and more preferably 4 ° C.
  • a fermented soybean whey beverage prepared by the above-described method for preparing a fermented soybean whey beverage.
  • the fermented soybean whey beverage of the invention has low sucrose content, high content of soybean oligosaccharide active ingredients such as stachyose and raffinose, can effectively promote the proliferation of human intestinal bifidobacteria, increase beneficial bacteria, inhibit harmful bacteria, and moisten
  • the bowels are laxative, remove toxins, and improve the intestinal immune function of the human body.
  • Soybean meal was pulverized, sieved, and mixed with deionized water at a ratio of 1:13 by a conventional alkali-soluble acid precipitation method.
  • the pH was adjusted to 8.5 with a NaOH solution, and the mixture was stirred at a constant temperature of 250 ° C for 50 minutes at 50 ° C; the pH was adjusted with a HCl solution. 4.5, 40 ° C constant temperature, 250r / min stirred for 20min, centrifugation, the supernatant was concentrated by evaporation on a rotary evaporator 50 ° C, 80kPa pressure, retaining concentrated whey concentration of 10% whey.
  • the above-mentioned concentrated soybean whey is compounded with milk powder, and the raw material for fermentation is often sterilized by temperature and high pressure, and the amount of milk powder added is 5% of the quality of the raw material for fermentation.
  • the sucrose was replaced by glucose in the MRS medium to prepare a No. 1 MRS medium containing no glucose.
  • the sucrose and soybean oligosaccharide were mixed in a ratio of 1:1 to replace the glucose in the MRS medium to prepare a No. 2 MRS medium containing no glucose.
  • the lyophilized lactic acid bacteria were picked and inoculated on the No. 1 MRS agar plate medium containing no glucose. After anaerobic incubation for 24 hours at 38 °C, a single colony was picked in 5 mL of glucose-free MRS liquid medium. The amount of sucrose and soybean oligosaccharide residual sugar was measured after anaerobic incubation at 38 °C for 24 h.
  • lactic acid bacteria with sucrose residual sugar rate of 1% or less and soybean oligosaccharide residual sugar rate of 95% or higher, anaerobic culture at 38 °C until the logarithmic phase of growth take 200 ⁇ L of bacterial liquid, and transfer to sterile water to prepare the bacterial cell concentration.
  • Sterile sodium alginate was added to the lactic acid bacteria suspension selectively using sucrose.
  • the amount of sterile sodium alginate was 2% of the mass of the lactic acid bacteria suspension. After mixing, the mixture was dropped into CaCl 2 at a concentration of 2%.
  • the immobilized lactic acid bacteria colloidal particles having a diameter of 2 mm were prepared by standing and washing with sterile physiological saline.
  • the immobilized lactic acid bacteria colloidal particles and the fermentation raw material are mixed at a ratio of 1:3, 0.2wt% anhydrous sodium acetate is added to the initial fermentation raw material, and the fermentation is carried out at 38 ° C for 24 hours, and then filtered, and the immobilized lactic acid bacteria colloidal particles are removed, and the temperature is often high and high.
  • the soybean whey beverage is prepared; the filtered immobilized lactic acid bacteria colloidal particles are washed with sterile physiological saline and stored at 4 ° C in physiological saline for use.
  • Soybean meal was pulverized, sieved, and mixed with deionized water at a ratio of 1:11 by a conventional alkali-soluble acid precipitation method.
  • the pH was adjusted to 8.5 with a NaOH solution, and the mixture was stirred at a constant temperature of 250 ° C for 50 minutes at 50 ° C; the pH was adjusted with HCl solution. 4.5, 40 ° C constant temperature, 250r / min stirred for 20min, centrifugation, the supernatant was concentrated by evaporation on a rotary evaporator 70 ° C, 90kPa pressure, retaining whey concentration of 12% concentrated soy whey.
  • the concentrated soybean whey and the goat milk powder are compounded, and the fermentation raw material is often sterilized by temperature and high pressure, and the milk powder is added in an amount of 10% of the quality of the fermentation raw material.
  • the sucrose was replaced by glucose in the MRS medium to prepare a No. 1 MRS medium containing no glucose.
  • the sucrose and soybean oligosaccharide were mixed in a ratio of 1:1 to replace the glucose in the MRS medium to prepare a No. 2 MRS medium containing no glucose.
  • the lyophilized lactic acid bacteria strains were picked and inoculated on the No. 1 MRS agar plate medium containing no glucose. After anaerobic incubation for 36 hours at 37 °C, single colonies were picked in 5 mL of glucose-free MRS liquid medium. The amount of sucrose and soybean oligosaccharide residual sugar was measured after anaerobic incubation at 37 °C for 36 h.
  • the immobilized lactic acid bacteria colloidal particles having a diameter of 2.5 mm were prepared by standing and washing with sterile physiological saline.
  • the immobilized lactic acid bacteria colloidal particles and the fermentation starting raw material are mixed at a ratio of 1:3, 0.3 wt% anhydrous sodium acetate is added to the initial fermentation raw material, and the mixture is heated at 37 ° C for 36 h, and then filtered, and the immobilized lactic acid bacteria colloidal particles are removed, and the temperature is often high and high.
  • the soybean whey beverage is prepared; the filtered immobilized lactic acid bacteria colloidal particles are washed with sterile physiological saline and stored at 4 ° C in physiological saline for use.
  • Soybean meal was pulverized, sieved, and mixed with deionized water at a ratio of 1:10 by a conventional alkali-soluble acid precipitation method.
  • the pH was adjusted to 8.5 with a NaOH solution, and the mixture was stirred at a constant temperature of 50 ° C for 50 minutes at 250 ° C.
  • the pH was adjusted with a HCl solution. 4.5, 40 ° C constant temperature, 250r / min stirred for 20min, centrifugation, the supernatant was evaporated by a rotary evaporator at 55 ° C, 83kPa pressure, retaining concentrated whey with a whey concentration of 13%.
  • the concentrated soybean whey and the milk powder are compounded, and the fermentation raw material is often sterilized by temperature and high pressure, and the goat milk powder is added in an amount of 15% of the quality of the fermentation raw material.
  • the sucrose was replaced by glucose in the MRS medium to prepare a No. 1 MRS medium containing no glucose.
  • the sucrose and soybean oligosaccharide were mixed in a ratio of 1:1 to replace the glucose in the MRS medium to prepare a No. 2 MRS medium containing no glucose.
  • the lyophilized lactic acid bacteria strains were picked and inoculated on No. 1 MRS agar plate medium containing no glucose. After anaerobic incubation for 48 hours at 40 °C, single colonies were picked in 5 mL of MRS-free medium containing no glucose. The amount of sucrose and soybean oligosaccharide residual sugar was measured after anaerobic culture for 48 hours at 40 °C.
  • lactic acid bacteria with sucrose residual sugar content of 1% or less and soybean oligosaccharide residual sugar rate of 95% or higher, anaerobic culture at 40 °C until the logarithmic phase of growth take 200 ⁇ L of bacterial liquid, and transfer to sterile water to prepare the bacterial cell concentration.
  • Sterile sodium alginate was added to the lactic acid bacteria suspension selectively using sucrose. The amount of sterile sodium alginate was 2.5% of the mass of the lactic acid bacteria suspension. After mixing, the mixture was dropped into CaCl 2 at a concentration of 4%.
  • the immobilized lactic acid bacteria colloidal particles having a diameter of 3 mm were prepared by standing and washing with sterile physiological saline.
  • the immobilized lactic acid bacteria colloidal particles and the fermentation raw material are mixed at a ratio of 1:4, 0.5 wt% anhydrous sodium acetate is added to the initial fermentation raw material, and the fermentation is carried out at 40 ° C for 48 h, and then filtered, and the immobilized lactic acid bacteria colloidal particles are removed, and the temperature is often high and high.
  • the soybean whey beverage is prepared; the filtered immobilized lactic acid bacteria colloidal particles are washed with sterile physiological saline and stored at 4 ° C in physiological saline for use.
  • Soybean meal was pulverized, sieved, and mixed with deionized water by 1:8, adjusted to pH 8.5 with NaOH solution, stirred at 50 ° C for 50 min at 250 ° C, and adjusted to pH with HCl solution. 4.5, 40 ° C constant temperature, 250r / min stirred for 20min, centrifugation, the supernatant was concentrated by evaporation on a rotary evaporator 65 ° C, 87kPa pressure, retaining whey concentration of 15% concentrated soy whey.
  • the concentrated soybean whey and the goat milk powder are compounded, and the fermentation raw material is often sterilized by temperature and high pressure, and the goat milk powder is added in an amount of 20% of the quality of the fermentation raw material.
  • the sucrose was replaced by glucose in the MRS medium to prepare a No. 1 MRS medium containing no glucose.
  • the sucrose and soybean oligosaccharide were mixed in a ratio of 1:1 to replace the glucose in the MRS medium to prepare a No. 2 MRS medium containing no glucose.
  • the lyophilized lactic acid bacteria strains were picked and inoculated on No. 1 MRS agar plate medium containing no glucose. After anaerobic incubation for 24 hours at 42 °C, single colonies were picked in 5 mL of glucose-free MRS liquid medium. The amount of sucrose and soybean oligosaccharide residual sugar was measured after anaerobic culture for 24 hours at 42 °C.
  • the immobilized lactic acid bacteria colloidal particles having a diameter of 3 mm were prepared by standing and washing with sterile physiological saline.
  • the immobilized lactic acid bacteria colloidal particles and the fermentation raw material are mixed at a ratio of 1:4, 0.6 wt% anhydrous sodium acetate is added to the initial fermentation raw material, and the fermentation is carried out at 42 ° C for 24 h, and then filtered to remove the immobilized lactic acid bacteria colloidal particles, and the temperature is usually high and high.
  • the soybean whey beverage is prepared; the filtered immobilized lactic acid bacteria colloidal particles are washed with sterile physiological saline and stored at 4 ° C in physiological saline for use.
  • Soybean meal was pulverized, sieved, and mixed with deionized water at a ratio of 1:10 by a conventional alkali-soluble acid precipitation method.
  • the pH was adjusted to 8.5 with a NaOH solution, and the mixture was stirred at a constant temperature of 50 ° C for 50 minutes at 250 ° C.
  • the pH was adjusted with a HCl solution. 4.5, 40 ° C constant temperature, 250r / min stirred for 20min, centrifugation, the supernatant was evaporated by a rotary evaporator at 60 ° C, 85kPa pressure, retaining concentrated whey with a whey concentration of 13%.
  • the above-mentioned concentrated soybean whey is compounded with goat milk powder, and the raw material for fermentation is often sterilized by temperature and high pressure, and the amount of milk powder added is 15% of the quality of the raw material for fermentation.
  • the sucrose was replaced by glucose in the MRS medium to prepare a No. 1 MRS medium containing no glucose.
  • the sucrose and soybean oligosaccharide were mixed in a ratio of 1:1 to replace the glucose in the MRS medium to prepare a No. 2 MRS medium containing no glucose.
  • the lyophilized lactic acid bacteria strains were picked and inoculated on No. 1 MRS agar plate medium containing no glucose. After anaerobic incubation for 36 hours at 39 °C, single colonies were picked in 5 mL of glucose-free MRS liquid medium. The amount of sucrose and soybean oligosaccharide residual sugar was measured after anaerobic incubation at 39 °C for 36 h.
  • the immobilized lactic acid bacteria colloidal particles having a diameter of 2.5 mm were prepared by standing and washing with sterile physiological saline.
  • the immobilized lactic acid bacteria colloidal particles and the fermentation raw material are mixed at a ratio of 1:3.5, 0.4 wt% anhydrous sodium acetate is added to the initial fermentation raw material, and the fermentation is carried out at 39 ° C for 36 h, and then filtered, and the immobilized lactic acid bacteria colloidal particles are removed, and the temperature is often high and high.
  • the soybean whey beverage is prepared; the filtered immobilized lactic acid bacteria colloidal particles are washed with sterile physiological saline and stored at 4 ° C in physiological saline for use.
  • sucrose and soybean oligosaccharide content in the obtained soybean whey beverage was determined by HPLC, and the results are shown in Table 1:
  • Example Sucrose content wt% Soybean oligosaccharide content wt% Example 1 0.7 4.5
  • Example 2 0.6 4.9
  • Example 3 0.4 5.3
  • Example 4 0.6 5.7
  • Example 5 0.4 5.8
  • the soybean whey beverage prepared by the method of the present invention has a sucrose content of 0.7 wt% or less and a soybean oligosaccharide content of 4.5 wt% or more, and the soybean of the health beverage product obtained by the patent 201210393907.X
  • the highest content of oligosaccharides does not exceed 1.1 wt%, indicating that the method of the invention effectively removes sucrose from soybean whey, retains the active components of soybean oligosaccharides such as stachyose and raffinose; and the large preparation prepared by the method of the invention
  • Soy whey beverage can effectively promote the proliferation of human intestinal bifidobacteria, increase beneficial bacteria, inhibit harmful bacteria, laxative, remove toxins, and improve human intestinal immune function.
  • Example Reusable times / times Example 1 10
  • Example 2 12 Example 3 15
  • Example 4 15 Example 5 15
  • the immobilized lactic acid bacteria colloidal particles obtained by the present invention can be reused for 10-15 times, and the present invention adopts the microbial immobilization fermentation method to efficiently utilize the soybean industrial by-products while maintaining the catalysis of the lactic acid bacteria in the fermentation liquid.
  • the activity can also recycle the immobilized lactic acid bacteria after the continuous fermentation reaction, and the process is simple and the cost is low.

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Abstract

一种发酵型大豆乳清饮料及其制备方法，所述发酵型大豆乳清饮料的制备方法采用固定化乳酸菌胶进行发酵，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌，工艺简单，成本低廉，所述发酵型大豆乳清饮料中蔗糖含量低，水苏糖、棉子糖等大豆低聚糖活性成分含量高，能有效促进人体肠道双歧杆菌增殖，增加有益菌、抑制有害菌、润肠通便、清除毒素、提高人体肠道免疫功能。

Description

一种发酵型大豆乳清饮料及其制备方法
本申请要求于2016年12月7日提交中国专利局、申请号为2016111178628、名称为“一种发酵型大豆乳清饮料及其制备方法”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。
技术领域
本发明涉及食品加工方法技术领域，具体而言，涉及一种发酵型大豆乳清饮料及其制备方法。
背景技术
大豆乳清是大豆蛋白加工过程副产物，其富含乳清蛋白、大豆低聚糖、无机盐等，目前多以废物直接排放，造成资源浪费，引起环境污染。大豆乳清中的大豆低聚糖是一种功能性糖类，其主要成分为蔗糖、水苏糖和棉子糖，除蔗糖外，其余基本不被人体吸收，可直达肠道，促进肠道双歧杆菌增殖，可抑制有害菌，增进有益菌，润肠通便，清除毒素，提高肠道免疫功能。蔗糖是一种常见的双糖，摄入过多容易引起肥胖、蛀牙，对糖尿病患者也极为不利。
为除去蔗糖，提高功能性低聚糖纯度，现有技术采用了很多方法，如专利201510899907.0公开了一种大豆深加工过程中低聚糖的提取方法，其对大豆乳清除杂预处理后，经超滤、纳滤、喷雾干燥得到高纯度低聚糖，提高了大豆加工产业附加值；专利201210393907.X公开了一种以黄浆水为主要原料的保健饮料及其制作方法，其以大豆黄浆水为主要原料，添加蔗糖、果汁、酸味剂、食用香精以及豆渣发酵液制成保健饮料，该方法在保 留大豆低聚糖基础上，充分利用豆制品副产物，解决了黄浆水和豆渣的处理问题；
然而现有技术方法的缺点较多，如专利201510899907.0提纯方法低聚糖纯度不高、过程繁琐、不适宜大规模生产，而且能耗较高、成本较大，还不可避免的产生部分低聚糖成分的损失；按专利201210393907.X方法制作的产品中所含生物活性物质，如大豆低聚糖含量偏低，效果有限。
有鉴于此，特提出本发明。
发明内容
本发明的第一目的在于提供一种发酵型大豆乳清饮料的制备方法，所述的发酵型大豆乳清饮料的制备方法在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌，工艺简单，成本低廉；此外，有效去除大豆乳清中的蔗糖，充分保留水苏糖、棉子糖等大豆低聚糖活性成分。
本发明的第二目的在于提供一种采用上述的发酵型大豆乳清饮料的制备方法制备得到的发酵型大豆乳清饮料，所述的发酵型大豆乳清饮料中蔗糖含量低，水苏糖、棉子糖等大豆低聚糖活性成分含量高，能有效促进人体肠道双歧杆菌增殖，增加有益菌、抑制有害菌、润肠通便、清除毒素、提高人体肠道免疫功能。
为了实现本发明的上述目的，特采用以下技术方案：
一种发酵型大豆乳清饮料的制备方法，以大豆乳清为原料，依次经浓缩、复配后与固定化乳酸菌胶粒混合发酵制得大豆乳清饮料。
本发明发酵型大豆乳清饮料的制备方法有效克服了现有的大豆乳清中大豆低聚糖等活性成分提取技术落后，所得产品中大豆低聚糖等活性成 分含量低，大豆乳清相关产品生产过程能耗较高、成本较大、产品功效不明确等缺点或不足，本发明采用固定化乳酸菌胶进行发酵，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌，工艺简单，成本低廉。
优选地，所述浓缩包括将大豆乳清加热加压蒸发浓缩至大豆乳清浓度为10wt％-15wt％，优选为12wt％-13wt％，进一步优选为13wt％。
优选地，所述加热温度为50-70℃，优选为55-65℃，进一步优选为60℃。
优选地，所述加压压力为80-90kPa，优选为83-87kPa，进一步优选为85kPa。
优选地，所述复配包括：将浓缩后的大豆乳清与奶粉复配，经灭菌得发酵初始原料，其中奶粉添加量为所得发酵初始原料质量的5％-20％，优选为10％-20％，进一步优选为15％。
优选地，所述固定化乳酸菌胶粒的制备方法包括：将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，混匀后，将所得混合液加入CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为2-3mm，优选为2.5-3mm，进一步优选为2.5mm的固定化乳酸菌胶粒。
优选地，所述无菌海藻酸钠的加入量为乳酸菌悬液质量的2％-2.5％，优选为2.3％-2.5％，进一步优选为2.3％。
优选地，所述CaCl2溶液的浓度为2％-4％，优选为3％-4％，进一步优选为3％。
优选地，所述发酵包括：将固定化乳酸菌胶粒与待发酵原料混合，加入乙酸钠，恒温发酵，过滤，去除固定化乳酸菌胶粒，灭菌即得大豆乳清饮料。
优选地，所述固定化乳酸菌胶粒与待发酵原料的质量比为1:3-4，优选为1:3.5-4，进一步优选为1:3.5。
优选地，所述乙酸钠的加入量为待发酵原料质量的0.2％-0.6％，优选为0.3％-0.5％，进一步优选为0.4％。
优选地，所述恒温发酵的发酵温度为37-42℃，优选为38-40℃，进一步优选为39℃；
优选地，所述恒温发酵的发酵时间为24-48h，优选为24-36h，进一步优选为36h。
优选地，所述过滤后所得固定化乳酸菌胶粒经无菌生理盐水清洗，在低温生理盐水中保存，重复利用。
采用上述的发酵型大豆乳清饮料的制备方法制备得到的一种发酵型大豆乳清饮料。
本发明发酵型大豆乳清饮料中蔗糖含量低，水苏糖、棉子糖等大豆低聚糖活性成分含量高，能有效促进人体肠道双歧杆菌增殖，增加有益菌、抑制有害菌、润肠通便、清除毒素、提高人体肠道免疫功能。
与现有技术相比，本发明的有益效果为：
本发明发酵型大豆乳清饮料的制备方法有效克服了现有的大豆乳清中大豆低聚糖等活性成分提取技术落后，所得产品中大豆低聚糖等活性成分含量低，大豆乳清相关产品生产过程能耗较高、成本较大、产品功效不明确等缺点或不足，本发明采用固定化乳酸菌胶进行发酵，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌，工艺简单，成本低廉。
本发明发酵型大豆乳清饮料中蔗糖含量低，水苏糖、棉子糖等大豆低聚糖活性成分含量高，能有效促进人体肠道双歧杆菌增殖，增加有益菌、抑制有害菌、润肠通便、清除毒素、提高人体肠道免疫功能。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案，下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图是本发明的一些实施方式，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为本发明一种具体实施方式提供的发酵型大豆乳清饮料的制备方法工艺流程图。
具体实施方式
下面将结合附图和具体实施方式对本发明的技术方案进行清楚、完整地描述，但是本领域技术人员将会理解，下列所描述的实施例是本发明一部分实施例，而不是全部的实施例，仅用于说明本发明，而不应视为限制本发明的范围。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。
图1为本发明一种具体实施方式提供的发酵型大豆乳清饮料的制备方法工艺流程图。
本发明提供了一种发酵型大豆乳清饮料的制备方法，以大豆乳清为原料，依次经浓缩、复配后与固定化乳酸菌胶粒混合发酵制得大豆乳清饮料。
本发明发酵型大豆乳清饮料的制备方法有效克服了现有的大豆乳清中大豆低聚糖等活性成分提取技术落后，所得产品中大豆低聚糖等活性成分含量低，大豆乳清相关产品生产过程能耗较高、成本较大、产品功效不明确等缺点或不足，本发明采用固定化乳酸菌胶进行发酵，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌，工艺简单，成本低廉。
优选地，所述大豆乳清可根据原料取材难易程度，选择源于工厂生产大豆分离蛋白时产生的大豆乳清副产物，或者以大豆豆粕为原料，采用传统碱溶酸沉法，将大豆豆粕粉碎、筛分、碱溶、酸沉、离心后得大豆乳清。
优选地，所述浓缩包括将大豆乳清加热加压蒸发浓缩至大豆乳清浓度为10wt％-15wt％，优选为12wt％-13wt％，进一步优选为13wt％。
在发酵前对大豆乳清进行适当浓缩，有助于提高后续发酵效率，促进蔗糖充分代谢，提高产品品质。
优选地，所述加热温度为50-70℃，优选为55-65℃，进一步优选为60℃。
优选地，所述加压压力为80-90kPa，优选为83-87kPa，进一步优选为85kPa。
采用特定温度和压力对大豆乳清进行蒸发浓缩，有助于提高浓缩效率。
优选地，所述蒸发浓缩采用旋转蒸发器进行。
优选地，所述复配包括：将浓缩后的大豆乳清与奶粉复配，经灭菌得发酵初始原料，其中奶粉添加量为所得发酵初始原料质量的5％-20％，优选为10％-20％，进一步优选为15％。
将浓缩后的大豆乳清与奶粉复配，一方面奶粉也提供了部分乳酸菌代谢原料，此外奶粉的加入，也可以提高所得发酵产品的营养价值，改善口感风味。
优选地，所述奶粉包括牛奶粉和/或羊奶粉。
优选地，所述奶粉包括全脂奶粉和/或脱脂奶粉，优选包括脱脂奶粉。
优选地，所述固定化乳酸菌胶粒的制备方法包括：将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，混匀后，将所得混合液加入CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为2-3mm，优选为2.5-3mm，进一步优选为2.5mm的固定化乳酸菌胶粒。
本发明采用特定方法，制备得到特定粒径大小的固定化乳酸菌胶粒，采用所得固定化乳酸菌胶进行发酵，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌。
优选地，所述无菌海藻酸钠的加入量为乳酸菌悬液质量的2％-2.5％，优选为2.3％-2.5％，进一步优选为2.3％。
无菌海藻酸钠可作为乳化稳定剂和增稠剂。
优选地，所述CaCl2溶液的浓度为2％-4％，优选为3％-4％，进一步优选为3％。
作为一种食品添加剂，氯化钙可起到多价螯合剂和固化剂的作用。
采用特定用量的无菌海藻酸钠和氯化钙，能够得到具有特定粒径尺寸的固定化乳酸菌胶粒。
进一步优选地，将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，混匀后，将所得混合液滴加入CaCl2溶液中。
优选地，所述乳酸菌悬液可采用自行培养、筛选、配置的方式得到。
进一步优选地，本发明具体实施方式所采用的乳酸菌悬液通过如下方法获得：
将蔗糖代替MRS培养基中葡萄糖，制成不含葡萄糖的1号MRS培养基。将蔗糖、大豆低聚糖以1:1混合，代替MRS培养基中葡萄糖，制成不含葡萄糖的2号MRS培养基。挑取冻干保藏的乳酸菌菌种，接种于不含葡萄糖的1号MRS琼脂平板培养基上，厌氧培养后，挑取单菌落于5mL不含葡萄糖的2号MRS液体培养基中，厌氧培养后检测蔗糖、大豆低聚糖残糖量。选择蔗糖残糖率为1％以下，大豆低聚糖残糖率为95％以上的乳酸菌，厌氧培养至生长对数期，取200μL菌液，移入无菌水中制成所需菌体浓度的活性乳酸菌悬液。
优选地，所述发酵包括：将固定化乳酸菌胶粒与待发酵原料(即上述的发酵初始原料)混合，加入乙酸钠，恒温发酵，过滤，去除固定化乳酸菌胶粒，灭菌即得大豆乳清饮料。
本发明采用固定化乳酸菌胶进行发酵，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌。
优选地，所述固定化乳酸菌胶粒与待发酵原料的质量比为1:3-4，优选为1:3.5-4，进一步优选为1:3.5。
采用特定用量比例的固定化乳酸菌胶粒与待发酵原料，有助于提高发酵效率，促进蔗糖充分代谢，提高产品品质。
优选地，所述乙酸钠的加入量为待发酵原料质量的0.2％-0.6％，优选为0.3％-0.5％，进一步优选为0.4％。
采用特定用量的乙酸钠，能够抑制其他杂菌的生长，保证乳酸菌的生长和增殖。
优选地，所述恒温发酵的发酵温度为37-42℃，优选为38-40℃，进一步优选为39℃。
优选地，所述恒温发酵的发酵时间为24-48h，优选为24-36h，进一步优选为36h。
采用特定发酵温度和发酵时间，有助于提高发酵效率，促进蔗糖充分代谢，提高产品品质。
优选地，所述过滤后所得固定化乳酸菌胶粒经无菌生理盐水清洗，在低温生理盐水中保存，重复利用。
优选地，所述低温为10℃以下，优选为5℃以下，进一步优选为4℃。
采用上述的发酵型大豆乳清饮料的制备方法制备得到的一种发酵型大豆乳清饮料。
本发明发酵型大豆乳清饮料中蔗糖含量低，水苏糖、棉子糖等大豆低聚糖活性成分含量高，能有效促进人体肠道双歧杆菌增殖，增加有益菌、抑制有害菌、润肠通便、清除毒素、提高人体肠道免疫功能。
实施例1
采用传统碱溶酸沉法，将大豆豆粕粉碎、筛分后与去离子水以1:13混合，用NaOH溶液调pH至8.5，50℃恒温、250r/min搅拌50min；用HCl溶液调pH至4.5，40℃恒温、250r/min搅拌20min，离心，上清液经旋转蒸发器50℃，80kPa压力蒸发浓缩，保留乳清浓度为10％的浓缩型大豆乳清。将上述浓缩大豆乳清与牛奶粉复配，经常温高压灭菌得发酵初始原料，牛奶粉添加量为发酵初始原料质量的5％。
将蔗糖代替MRS培养基中葡萄糖，制成不含葡萄糖的1号MRS培养基。将蔗糖、大豆低聚糖以1:1混合，代替MRS培养基中葡萄糖，制成不含葡萄糖的2号MRS培养基。挑取冻干保藏的乳酸菌菌种，接种于不含葡萄糖的1号MRS琼脂平板培养基上，38℃厌氧培养24h后，挑取单菌落于5mL 不含葡萄糖的2号MRS液体培养基中，38℃厌氧培养24h后检测蔗糖、大豆低聚糖残糖量。选择蔗糖残糖率为1％以下，大豆低聚糖残糖率为95％以上的乳酸菌，38℃厌氧培养至生长对数期，取200μL菌液，移入无菌水中制成菌体浓度为0.5×1010cfumL-1的选择性利用蔗糖的乳酸菌悬液。将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，无菌海藻酸钠的用量为乳酸菌悬液质量的2％，混匀后，将所得混合液滴入浓度为2％的CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为2mm的固定化乳酸菌胶粒。将固定化乳酸菌胶粒与发酵初始原料以1:3比例混合，加入占发酵初始原料0.2wt％无水乙酸钠，38℃恒温发酵24h后，过滤，去除固定化乳酸菌胶粒，并经常温高压二次灭菌，制备得到大豆乳清饮料；过滤后的固定化乳酸菌胶粒，经无菌生理盐水洗清，4℃生理盐水保存待用。
实施例2
采用传统碱溶酸沉法，将大豆豆粕粉碎、筛分后与去离子水以1:11混合，用NaOH溶液调pH至8.5，50℃恒温、250r/min搅拌50min；用HCl溶液调pH至4.5，40℃恒温、250r/min搅拌20min，离心，上清液经旋转蒸发器70℃，90kPa压力蒸发浓缩，保留乳清浓度为12％的浓缩型大豆乳清。将上述浓缩大豆乳清与羊奶粉复配，经常温高压灭菌得发酵初始原料，牛奶粉添加量为发酵初始原料质量的10％。
将蔗糖代替MRS培养基中葡萄糖，制成不含葡萄糖的1号MRS培养基。将蔗糖、大豆低聚糖以1:1混合，代替MRS培养基中葡萄糖，制成不含葡萄糖的2号MRS培养基。挑取冻干保藏的乳酸菌菌种，接种于不含葡萄糖的1号MRS琼脂平板培养基上，37℃厌氧培养36h后，挑取单菌落于5mL不含葡萄糖的2号MRS液体培养基中，37℃厌氧培养36h后检测蔗糖、大豆低聚糖残糖量。选择蔗糖残糖率为1％以下，大豆低聚糖残糖率为95％以上的乳酸菌，37℃厌氧培养至生长对数期，取200μL菌液，移入无菌水 中制成菌体浓度为0.7×1010cfumL-1的选择性利用蔗糖的乳酸菌悬液。将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，无菌海藻酸钠的用量为乳酸菌悬液质量的2％，混匀后，将所得混合液滴入浓度为3％的CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为2.5mm的固定化乳酸菌胶粒。将固定化乳酸菌胶粒与发酵初始原料以1:3比例混合，加入占发酵初始原料0.3wt％无水乙酸钠，37℃恒温发酵36h后，过滤，去除固定化乳酸菌胶粒，并经常温高压二次灭菌，制备得到大豆乳清饮料；过滤后的固定化乳酸菌胶粒，经无菌生理盐水洗清，4℃生理盐水保存待用。
实施例3
采用传统碱溶酸沉法，将大豆豆粕粉碎、筛分后与去离子水以1:10混合，用NaOH溶液调pH至8.5，50℃恒温、250r/min搅拌50min；用HCl溶液调pH至4.5，40℃恒温、250r/min搅拌20min，离心，上清液经旋转蒸发器55℃，83kPa压力蒸发浓缩，保留乳清浓度为13％的浓缩型大豆乳清。将上述浓缩大豆乳清与牛奶粉复配，经常温高压灭菌得发酵初始原料，羊奶粉添加量为发酵初始原料质量的15％。
将蔗糖代替MRS培养基中葡萄糖，制成不含葡萄糖的1号MRS培养基。将蔗糖、大豆低聚糖以1:1混合，代替MRS培养基中葡萄糖，制成不含葡萄糖的2号MRS培养基。挑取冻干保藏的乳酸菌菌种，接种于不含葡萄糖的1号MRS琼脂平板培养基上，40℃厌氧培养48h后，挑取单菌落于5mL不含葡萄糖的2号MRS液体培养基中，40℃厌氧培养48h后检测蔗糖、大豆低聚糖残糖量。选择蔗糖残糖率为1％以下，大豆低聚糖残糖率为95％以上的乳酸菌，40℃厌氧培养至生长对数期，取200μL菌液，移入无菌水中制成菌体浓度为0.99×1010cfumL-1的选择性利用蔗糖的乳酸菌悬液。将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，无菌海藻酸钠的用量为乳酸菌悬液质量的2.5％，混匀后，将所得混合液滴入浓度为4％的CaCl2 溶液中，静置、无菌生理盐水洗涤制得直径为3mm的固定化乳酸菌胶粒。将固定化乳酸菌胶粒与发酵初始原料以1:4比例混合，加入占发酵初始原料0.5wt％无水乙酸钠，40℃恒温发酵48h后，过滤，去除固定化乳酸菌胶粒，并经常温高压二次灭菌，制备得到大豆乳清饮料；过滤后的固定化乳酸菌胶粒，经无菌生理盐水洗清，4℃生理盐水保存待用。
实施例4
采用传统碱溶酸沉法，将大豆豆粕粉碎、筛分后与去离子水以1:8混合，用NaOH溶液调pH至8.5，50℃恒温、250r/min搅拌50min；用HCl溶液调pH至4.5，40℃恒温、250r/min搅拌20min，离心，上清液经旋转蒸发器65℃，87kPa压力蒸发浓缩，保留乳清浓度为15％的浓缩型大豆乳清。将上述浓缩大豆乳清与羊奶粉复配，经常温高压灭菌得发酵初始原料，羊奶粉添加量为发酵初始原料质量的20％。
将蔗糖代替MRS培养基中葡萄糖，制成不含葡萄糖的1号MRS培养基。将蔗糖、大豆低聚糖以1:1混合，代替MRS培养基中葡萄糖，制成不含葡萄糖的2号MRS培养基。挑取冻干保藏的乳酸菌菌种，接种于不含葡萄糖的1号MRS琼脂平板培养基上，42℃厌氧培养24h后，挑取单菌落于5mL不含葡萄糖的2号MRS液体培养基中，42℃厌氧培养24h后检测蔗糖、大豆低聚糖残糖量。选择蔗糖残糖率为1％以下，大豆低聚糖残糖率为95％以上的乳酸菌，42℃厌氧培养至生长对数期，取200μL菌液，移入无菌水中制成菌体浓度为1.1×1010cfumL-1的选择性利用蔗糖的乳酸菌悬液。将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，无菌海藻酸钠的用量为乳酸菌悬液质量的2.5％，混匀后，将所得混合液滴入浓度为4％的CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为3mm的固定化乳酸菌胶粒。将固定化乳酸菌胶粒与发酵初始原料以1:4比例混合，加入占发酵初始原料0.6wt％无水乙酸钠，42℃恒温发酵24h后，过滤，去除固定化乳酸菌胶粒， 并经常温高压二次灭菌，制备得到大豆乳清饮料；过滤后的固定化乳酸菌胶粒，经无菌生理盐水洗清，4℃生理盐水保存待用。
实施例5
采用传统碱溶酸沉法，将大豆豆粕粉碎、筛分后与去离子水以1:10混合，用NaOH溶液调pH至8.5，50℃恒温、250r/min搅拌50min；用HCl溶液调pH至4.5，40℃恒温、250r/min搅拌20min，离心，上清液经旋转蒸发器60℃，85kPa压力蒸发浓缩，保留乳清浓度为13％的浓缩型大豆乳清。将上述浓缩大豆乳清与羊奶粉复配，经常温高压灭菌得发酵初始原料，牛奶粉添加量为发酵初始原料质量的15％。
将蔗糖代替MRS培养基中葡萄糖，制成不含葡萄糖的1号MRS培养基。将蔗糖、大豆低聚糖以1:1混合，代替MRS培养基中葡萄糖，制成不含葡萄糖的2号MRS培养基。挑取冻干保藏的乳酸菌菌种，接种于不含葡萄糖的1号MRS琼脂平板培养基上，39℃厌氧培养36h后，挑取单菌落于5mL不含葡萄糖的2号MRS液体培养基中，39℃厌氧培养36h后检测蔗糖、大豆低聚糖残糖量。选择蔗糖残糖率为1％以下，大豆低聚糖残糖率为95％以上的乳酸菌，39℃厌氧培养至生长对数期，取200μL菌液，移入无菌水中制成菌体浓度为1.05×1010cfumL-1的选择性利用蔗糖的乳酸菌悬液。将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，无菌海藻酸钠的用量为乳酸菌悬液质量的2.3％，混匀后，将所得混合液滴入浓度为3％的CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为2.5mm的固定化乳酸菌胶粒。将固定化乳酸菌胶粒与发酵初始原料以1:3.5比例混合，加入占发酵初始原料0.4wt％无水乙酸钠，39℃恒温发酵36h后，过滤，去除固定化乳酸菌胶粒，并经常温高压二次灭菌，制备得到大豆乳清饮料；过滤后的固定化乳酸菌胶粒，经无菌生理盐水洗清，4℃生理盐水保存待用。
采用HPLC测定所得大豆乳清饮料中的蔗糖和大豆低聚糖含量，结果如表1所示：
表1 本发明大豆乳清饮料中的蔗糖和大豆低聚糖含量
实施例 蔗糖含量wt％ 大豆低聚糖含量wt％
实施例1 0.7 4.5
实施例2 0.6 4.9
实施例3 0.4 5.3
实施例4 0.6 5.7
实施例5 0.4 5.8
通过表1可以看出，采用本发明方法制备得到的大豆乳清饮料中蔗糖含量为0.7wt％以下，而大豆低聚糖含量为4.5wt％以上，而专利201210393907.X所得保健饮料产品中大豆低聚糖的最高含量不超过1.1wt％，说明本发明方法有效去除了大豆乳清中蔗糖，保留了水苏糖、棉子糖等大豆低聚糖活性成分；采用本发明方法制备得到的大豆乳清饮料能有效促进人体肠道双歧杆菌增殖，增加有益菌、抑制有害菌、润肠通便、清除毒素、提高人体肠道免疫功能。
本发明所得固定化乳酸菌胶粒可重复利用次数如表2所示：
表2 本发明固定化乳酸菌胶粒可重复利用次数
实施例 可重复利用次数/次
实施例1 10
实施例2 12
实施例3 15
实施例4 15
实施例5 15
通过表2可以看出，本发明所得固定化乳酸菌胶粒可重复利用10-15次，本发明采用微生物固定化发酵法，在高效利用大豆工业副产物的同时，既可保持发酵液中乳酸菌催化活性，又可在连续发酵反应后回收利用固定化乳酸菌，工艺简单，成本低廉。
尽管已用具体实施例来说明和描述了本发明，然而应意识到，以上各实施例仅用以说明本发明的技术方案，而非对其限制；本领域的普通技术人员应当理解：在不背离本发明的精神和范围的情况下，可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围；因此，这意味着在所附权利要求中包括属于本发明范围内的所有这些替换和修改。

Claims (10)

  1. 一种发酵型大豆乳清饮料的制备方法，其特征在于，以大豆乳清为原料，依次经浓缩、复配后与固定化乳酸菌胶粒混合发酵制得大豆乳清饮料。
  2. 根据权利要求1所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述浓缩包括将大豆乳清加热加压蒸发浓缩至大豆乳清浓度为10wt％-15wt％，优选为12wt％-13wt％，进一步优选为13wt％。
  3. 根据权利要求2所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述加热温度为50-70℃，优选为55-65℃，进一步优选为60℃；
    优选地，所述加压压力为80-90kPa，优选为83-87kPa，进一步优选为85kPa。
  4. 根据权利要求1所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述复配包括：将浓缩后的大豆乳清与奶粉复配，经灭菌得发酵初始原料，其中奶粉添加量为所得发酵初始原料质量的5％-20％，优选为10％-20％，进一步优选为15％。
  5. 根据权利要求1所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述固定化乳酸菌胶粒的制备方法包括：将无菌海藻酸钠加入选择性利用蔗糖的乳酸菌悬液中，混匀后，将所得混合液加入CaCl2溶液中，静置、无菌生理盐水洗涤制得直径为2-3mm，优选为2.5-3mm，进一步优选为2.5mm的固定化乳酸菌胶粒。
  6. 根据权利要求5所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述无菌海藻酸钠的加入量为乳酸菌悬液质量的2％-2.5％，优选为2.3％-2.5％，进一步优选为2.3％；
    优选地，所述CaCl2溶液的浓度为2％-4％，优选为3％-4％，进一步优选为3％。
  7. 根据权利要求1所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述发酵包括：将固定化乳酸菌胶粒与待发酵原料混合，加入乙酸钠，恒温发酵，过滤，去除固定化乳酸菌胶粒，灭菌即得大豆乳清饮料。
  8. 根据权利要求7所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述固定化乳酸菌胶粒与待发酵原料的质量比为1:3-4，优选为1:3.5-4，进一步优选为1:3.5；
    优选地，所述乙酸钠的加入量为待发酵原料质量的0.2％-0.6％，优选为0.3％-0.5％，进一步优选为0.4％；
    优选地，所述恒温发酵的发酵温度为37-42℃，优选为38-40℃，进一步优选为39℃；
    优选地，所述恒温发酵的发酵时间为24-48h，优选为24-36h，进一步优选为36h。
  9. 根据权利要求7所述的一种发酵型大豆乳清饮料的制备方法，其特征在于，所述过滤后所得固定化乳酸菌胶粒经无菌生理盐水清洗，在低温生理盐水中保存，重复利用。
  10. 采用权利要求1-9任一所述的发酵型大豆乳清饮料的制备方法制备得到的一种发酵型大豆乳清饮料。
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