(3) summary of the invention
The object of the invention is to provide a kind of preparation method of probiotic fermented rice milk.
The object of the present invention is achieved like this: related percentage is weight ratio except other has indicating among the present invention, and product of the present invention adopts such method to prepare:
1. the selection of enzyme preparation
High temperature resistant AMS, unit of activity 20,000 u/g; Carbohydrase, unit of activity 50,000 u/g; Papain, unit of activity 500,000 u/g.
2. the preparation of the selection of probio and culture medium
A bacterium---Lactobacillus rhamnosus CICC6162 (Lactobacillus rhamnosus), culture medium is: peptone 2g, yeast extract 1g, soy peptone 1g, protein digestion thing No.3 (Difco) 1g, 5% ironic citrate 2ml, artificial seawater 700ml, distilled water 300ml, pH 7.5, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
B bacterium---lactobacillus paracasei class cheese subspecies CICC6111 (Lactobacillus paracasei subsp.paracasei), culture medium is: casein peptone 10g/L, meat soup extract 10g/L, yeast extract 5g/L, glucose 20g/L, Tween 80 1g/L, K
2HPO
42g/L, sodium acetate 5g/L, dibasic ammonium citrate 2g/L, MgSO
4* 7H
2O 0.2g/L, MnSO
4XH
2O 0.05g/L, pH 6.2-6.5, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
C bacterium---Lactobacillus brevis CICC20023 (Lac tobacillus brevis) culture medium is: yeast extract 7.5 grams, peptone 7.5 grams, glucose 10 grams, potassium dihydrogen phosphate 2 grams, 100 milliliters of tomato juices, 0.5 milliliter of Tween 80,900 milliliters in water, agar 1.5~2.0%, pH7.0, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
3. the preparation process of compound probiotic freeze-dried powder
3.1 former actication of culture
Measure respectively 0.9% physiological saline 10ml in 3 in vitro, be cooled to 30 ℃ in 20 minutes through 121 ℃ of sterilizations, freeze-dried vaccine powder in the 3 probiotics ampullas all poured under germ-free condition in 0.9% the physiological saline, concussion makes its dissolving, static activation is 30 minutes in 30 ℃ of insulating boxs, and is for subsequent use.
3.2 enlarging, probio cultivates
3.2.1 the preparation of mother culture
Measure respectively 3 each 200ml of probiotics culture medium in 3 500ml triangular flasks, 121 ℃ of sterilizations were cooled to 30 ℃ in 20 minutes, press 10% of culture volume and inoculate activated bacterial classification in 3.1 steps, static cultivation is 24 hours in 30 ℃ of anaerobic box, as mother culture.
3.2.2 produce the preparation of leavening
At first prepare mass percent concentration and be 10% defatted milk emulsion, be sub-packed in three 500mL triangular flasks, every bottle of 200mL, 121 ℃ of sterilizations were cooled to 30 ℃ in 20 minutes, respectively by 2% volume ratio inoculation mother culture, 30 ℃ of anaerobism cultivate 30---36 hours, detect 3 probiotics zymotic fluid viable counts, each zymotic fluid viable count 〉=10
9Individual/ml, treat as and be fermenting-ripening, if viable count<10
9Individual/ml, continue to cultivate, until reach 10
9Individual/ml.
3.3 the preparation of powder freeze-drying lactobacillus
The production leavening of maturation is imported in the glass ampoule under aseptic condition, and liquid level is lower than 1cm, puts into-30 ℃ of refrigerator-freezer quick-frozens after adding a cover bottle stopper, after freezing glass ampoule is taken advantage of dress with pallet, puts into freeze dryer and carries out freeze drying.
3.4 the compound probiotic freeze-dried powder is composite
Press the freeze-dried vaccine powder weight, get A bacterium powder 2-4 part, B bacterium powder 3-5 part, C bacterium powder 1-2 part fully is used for rice and brown rice rice breast mixed slurry behind the fermentation enzymolysis behind the mixing, add the bacterium amount and be 0.01% of rice behind the enzymolysis and brown rice rice breast mixed slurry weight.
4. the preparation of probiotic fermented rice milk
4.1 the preparation of fermentation raw material
With rice, brown rice and the drinking water weight ratio feeding by 2: 1: 7, behind the colloid mill defibrination, cross 80 order dusting covers, make mixed with rice serum material.
4.2 batching
5% interpolation white granulated sugar by mixed with rice serum material weight heats while stirring, and white granulated sugar is fully dissolved, and makes to contain sugared mixed with rice serum material.
4.3 liquefaction
The sugared mixed with rice serum material that contains for preparing is squeezed into fermentation tank, Steam Heating feed liquid to 90 ℃, keep 20 minute by the 0.1% high temperature resistant AMS that adds unit of activity 20,000 u/g that contains sugared mixed with rice serum material weight this moment, constantly stir, then be cooled to 50 ℃-55 ℃.
4.4 saccharification
By contain sugared mixed with rice serum material weight 0.15% to add unit of activity be 50,000 u/g carbohydrase, again by contain sugared mixed with rice serum material weight 0.2% to add unit of activity be 500,000 u/g papains, after stirring, 50 ℃-55 ℃ kept 4 hours.
The enzyme 4.5 go out
To be heated to through the feed liquid of 4.4 step process 100 ℃, keep 15 minutes, then be cooled to 30 ℃.
4.6 composite probiotics ferment
Add the compound probiotic freeze-dried powder that 3.4 steps prepare by 0.01% of the feed liquid weight after 4.5 step process, stir, 30 ℃ are incubated static fermentation 4 hours.
4.7 sterile filling
After the feed liquid of 4.6 step process stirred, carry out sterile filling, obtain the probiotic fermented rice milk finished product.
(4) specific embodiment
For a more detailed description to the present invention below in conjunction with specific embodiment:
Embodiment one:
3 probiotics that the present invention adopts are all purchased in Chinese industrial microorganism fungus kind preservation center, be respectively: Lactobacillus rhamnosus CICC6162, lactobacillus paracasei class cheese subspecies CICC6111, Lactobacillus brevis CICC20023, among the present invention with above-mentioned bacterial strains referred to as A bacterium, B bacterium, C bacterium.
1. the selection of enzyme preparation
High temperature resistant AMS, unit of activity 20,000 u/g; Carbohydrase, unit of activity 50,000 u/g; Papain, unit of activity 500,000 u/g.
2. the preparation of the selection of probio and culture medium
A bacterium---Lactobacillus rhamnosus CICC6162 (Lactobacillus rhamnosus), culture medium is: peptone 2g, yeast extract 1g, soy peptone 1g, protein digestion thing No.3 (Difco) 1g, 5% ironic citrate 2ml, artificial seawater 700ml, distilled water 300ml, pH 7.5, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
B bacterium---lactobacillus paracasei class cheese subspecies CICC6111 (Lactobacillus paracasei subsp.paracasei), culture medium is: casein peptone 10g/L, meat soup extract 10g/L, yeast extract 5g/L, glucose 20g/L, Tween 80 1g/L, K
2HPO
42g/L, sodium acetate 5g/L, dibasic ammonium citrate 2g/L, MgSO
4* 7H
2O 0.2g/L, MnSO
4XH
2O 0.05g/L, pH 6.2-6.5, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
C bacterium---Lactobacillus brevis CICC20023 (Lactobacillus brevis) culture medium is: yeast extract 7.5 grams, peptone 7.5 grams, glucose 10 grams, potassium dihydrogen phosphate 2 grams, 100 milliliters of tomato juices, 0.5 milliliter of Tween 80,900 milliliters in water, agar 1.5~2.0%, pH7.0, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
3. the preparation process of compound probiotic freeze-dried powder
3.1 former actication of culture
Measure respectively 0.9% physiological saline 10ml in 3 in vitro, be cooled to 30 ℃ in 20 minutes through 121 ℃ of sterilizations, freeze-dried vaccine powder in the 3 probiotics ampullas all poured under germ-free condition in 0.9% the physiological saline, concussion makes its dissolving, static activation is 30 minutes in 30 ℃ of insulating boxs, and is for subsequent use.
3.2 enlarging, probio cultivates
3.2.1 the preparation of mother culture
Measure respectively 3 each 200ml of probiotics culture medium in 3 500ml triangular flasks, 121 ℃ of sterilizations were cooled to 30 ℃ in 20 minutes, press 10% of culture volume and inoculate activated bacterial classification in 3.1 steps, static cultivation is 24 hours in 30 ℃ of anaerobic box, as mother culture.
3.2.2 produce the preparation of leavening
At first prepare mass percent concentration and be 10% defatted milk emulsion, be sub-packed in three 500mL triangular flasks, every bottle of 200mL, 121 ℃ of sterilizations were cooled to 30 ℃ in 20 minutes, respectively by 2% volume ratio inoculation mother culture, 30 ℃ of anaerobism cultivate 30---36 hours, detect 3 probiotics zymotic fluid viable counts, each zymotic fluid viable count 〉=10
9Individual/ml, treat as and be fermenting-ripening, if viable count<10
9Individual/ml, continue to cultivate, until reach 10
9Individual/ml.
3.3 the preparation of powder freeze-drying lactobacillus
The production leavening of maturation is imported in the glass ampoule under aseptic condition, and liquid level is lower than 1cm, puts into-30 ℃ of refrigerator-freezer quick-frozens after adding a cover bottle stopper, after freezing glass ampoule is taken advantage of dress with pallet, puts into freeze dryer and carries out freeze drying.
3.4 the compound probiotic freeze-dried powder is composite
Press the freeze-dried vaccine powder weight, get 2 parts in A bacterium powder, 3 parts in B bacterium powder, 1 part in C bacterium powder fully is used for rice and brown rice rice breast mixed slurry behind the fermentation enzymolysis behind the mixing, add the bacterium amount and be 0.01% of rice behind the enzymolysis and brown rice rice breast mixed slurry weight.
4. the preparation of probiotic fermented rice milk
4.1 the preparation of fermentation raw material
With rice, brown rice and the drinking water weight ratio feeding by 2: 1: 7, behind the colloid mill defibrination, cross 80 order dusting covers, make mixed with rice serum material.
4.2 batching
5% interpolation white granulated sugar by mixed with rice serum material weight heats while stirring, and white granulated sugar is fully dissolved, and makes to contain sugared mixed with rice serum material.
4.3 liquefaction
The sugared mixed with rice serum material that contains for preparing is squeezed into fermentation tank, Steam Heating feed liquid to 90 ℃, keep 20 minute by the 0.1% high temperature resistant AMS that adds unit of activity 20,000 u/g that contains sugared mixed with rice serum material weight this moment, constantly stir, then be cooled to 50 ℃-55 ℃.
4.4 saccharification
By contain sugared mixed with rice serum material weight 0.15% to add unit of activity be 50,000 u/g carbohydrase, again by contain sugared mixed with rice serum material weight 0.2% to add unit of activity be 500,000 u/g papains, after stirring, 50 ℃-55 ℃ kept 4 hours.
The enzyme 4.5 go out
To be heated to through the feed liquid of 4.4 step process 100 ℃, keep 15 minutes, then be cooled to 30 ℃.
4.6 composite probiotics ferment
Add the compound probiotic freeze-dried powder that 3.4 steps prepare by 0.01% of the feed liquid weight after 4.5 step process, stir, 30 ℃ are incubated static fermentation 4 hours.
4.7 sterile filling
After the feed liquid of 4.6 step process stirred, carry out sterile filling, obtain the probiotic fermented rice milk finished product.
Embodiment two:
1. the selection of enzyme preparation
High temperature resistant AMS, unit of activity 20,000 u/g; Carbohydrase, unit of activity 50,000 u/g; Papain, unit of activity 500,000 u/g.
2. the preparation of the selection of probio and culture medium
A bacterium---Lactobacillus rhamnosus CICC6162 (Lactobacillus rhamnosus), culture medium is: peptone 2g, yeast extract 1g, soy peptone 1g, protein digestion thing No.3 (Difco) 1g, 5% ironic citrate 2ml, artificial seawater 700ml, distilled water 300ml, pH 7.5, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
B bacterium---lactobacillus paracasei class cheese subspecies CICC6111 (Lactobacillus paracasei subsp.paracasei), culture medium is: casein peptone 10g/L, meat soup extract 10g/L, yeast extract 5g/L, glucose 20g/L, Tween 80 1g/L, K
2HPO
42g/L, sodium acetate 5g/L, dibasic ammonium citrate 2g/L, MgSO
4* 7H
2O 0.2g/L, MnSO
4XH
2O 0.05g/L, pH 6.2-6.5, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
C bacterium---Lactobacillus brevis CICC20023 (Lactobacillus brevis) culture medium is: yeast extract 7.5 grams, peptone 7.5 grams, glucose 10 grams, potassium dihydrogen phosphate 2 grams, 100 milliliters of tomato juices, 0.5 milliliter of Tween 80,900 milliliters in water, agar 1.5~2.0%, pH7.0, through 121 ℃ of sterilizations be cooled in 20 minutes 35 ℃ for subsequent use.
3. the preparation process of compound probiotic freeze-dried powder
3.1 former actication of culture
Measure respectively 0.9% physiological saline 10ml in 3 in vitro, be cooled to 30 ℃ in 20 minutes through 121 ℃ of sterilizations, freeze-dried vaccine powder in the 3 probiotics ampullas all poured under germ-free condition in 0.9% the physiological saline, concussion makes its dissolving, static activation is 30 minutes in 30 ℃ of insulating boxs, and is for subsequent use.
3.2 enlarging, probio cultivates
3.2.1 the preparation of mother culture
Measure respectively 3 each 200ml of probiotics culture medium in 3 500ml triangular flasks, 121 ℃ of sterilizations were cooled to 30 ℃ in 20 minutes, press 10% of culture volume and inoculate activated bacterial classification in 3.1 steps, static cultivation is 24 hours in 30 ℃ of anaerobic box, as mother culture.
3.2.2 produce the preparation of leavening
At first prepare mass percent concentration and be 10% defatted milk emulsion, be sub-packed in three 500mL triangular flasks, every bottle of 200mL, 121 ℃ of sterilizations were cooled to 30 ℃ in 20 minutes, respectively by 2% volume ratio inoculation mother culture, 30 ℃ of anaerobism cultivate 30---36 hours, detect 3 probiotics zymotic fluid viable counts, each zymotic fluid viable count 〉=10
9Individual/ml, treat as and be fermenting-ripening, if viable count<10
9Individual/ml, continue to cultivate, until reach 10
9Individual/ml.
3.3 the preparation of powder freeze-drying lactobacillus
The production leavening of maturation is imported in the glass ampoule under aseptic condition, and liquid level is lower than 1cm, puts into-30 ℃ of refrigerator-freezer quick-frozens after adding a cover bottle stopper, after freezing glass ampoule is taken advantage of dress with pallet, puts into freeze dryer and carries out freeze drying.
3.4 the compound probiotic freeze-dried powder is composite
Press the freeze-dried vaccine powder weight, get 4 parts in A bacterium powder, 5 parts in B bacterium powder, 2 parts in C bacterium powder fully is used for rice and brown rice rice breast mixed slurry behind the fermentation enzymolysis behind the mixing, add the bacterium amount and be 0.01% of rice behind the enzymolysis and brown rice rice breast mixed slurry weight.
4. the preparation of probiotic fermented rice milk
4.1 the preparation of fermentation raw material
With rice, brown rice and the drinking water weight ratio feeding by 2: 1: 7, behind the colloid mill defibrination, cross 80 order dusting covers, make mixed with rice serum material.
4.2 batching
5% interpolation white granulated sugar by mixed with rice serum material weight heats while stirring, and white granulated sugar is fully dissolved, and makes to contain sugared mixed with rice serum material.
4.3 liquefaction
The sugared mixed with rice serum material that contains for preparing is squeezed into fermentation tank, Steam Heating feed liquid to 90 ℃, keep 20 minute by the 0.1% high temperature resistant AMS that adds unit of activity 20,000 u/g that contains sugared mixed with rice serum material weight this moment, constantly stir, then be cooled to 50 ℃-55 ℃.
4.4 saccharification
By contain sugared mixed with rice serum material weight 0.15% to add unit of activity be 50,000 u/g carbohydrase, again by contain sugared mixed with rice serum material weight 0.2% to add unit of activity be 500,000 u/g papains, after stirring, 50 ℃-55 ℃ kept 4 hours.
The enzyme 4.5 go out
To be heated to through the feed liquid of 4.4 step process 100 ℃, keep 15 minutes, then be cooled to 30 ℃.
4.6 composite probiotics ferment
Add the compound probiotic freeze-dried powder that 3.4 steps prepare by 0.01% of the feed liquid weight after 4.5 step process, stir, 30 ℃ are incubated static fermentation 4 hours.
4.7 sterile filling
After the feed liquid of 4.6 step process stirred, carry out sterile filling, obtain the probiotic fermented rice milk finished product.