CN105661230A - Probiotic functional beverage and probiotic food prepared from Lens culinaris by fermentation - Google Patents
Probiotic functional beverage and probiotic food prepared from Lens culinaris by fermentation Download PDFInfo
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- CN105661230A CN105661230A CN201610036656.8A CN201610036656A CN105661230A CN 105661230 A CN105661230 A CN 105661230A CN 201610036656 A CN201610036656 A CN 201610036656A CN 105661230 A CN105661230 A CN 105661230A
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- Prior art keywords
- fermentation
- radix crotalariae
- crotalariae szemoensis
- lactobacillus plantarum
- solid
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- 230000004151 fermentation Effects 0.000 title claims abstract description 147
- 238000000855 fermentation Methods 0.000 title claims abstract description 146
- 239000006041 probiotic Substances 0.000 title claims abstract description 46
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 46
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 37
- 235000013305 food Nutrition 0.000 title abstract description 22
- 240000004322 Lens culinaris Species 0.000 title abstract 5
- 235000010666 Lens esculenta Nutrition 0.000 title abstract 5
- 235000020510 functional beverage Nutrition 0.000 title abstract 2
- 239000007788 liquid Substances 0.000 claims abstract description 49
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 28
- 235000013361 beverage Nutrition 0.000 claims abstract description 20
- 238000002360 preparation method Methods 0.000 claims abstract description 7
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 86
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 83
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 83
- 241000894006 Bacteria Species 0.000 claims description 79
- 241000186000 Bifidobacterium Species 0.000 claims description 66
- 241001465754 Metazoa Species 0.000 claims description 66
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 48
- 239000001963 growth medium Substances 0.000 claims description 40
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
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- 238000002474 experimental method Methods 0.000 description 3
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- 230000036541 health Effects 0.000 description 3
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- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical group OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
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- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/515—Animalis
Landscapes
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to the field of functional foods, particularly relates to a probiotic functional beverage and a probiotic food prepared from Lens culinaris by fermentation and discloses a liquid-state probiotic fermentation beverage prepared from Lens culinaris by liquid-state fermentation of a probiotic liquid mixture containing multiple kinds of probiotics, a probiotic food prepared from Lens culinaris by solid-state fermentation of a probiotic liquid mixture containing multiple kinds of probiotics and preparation methods of the liquid-state probiotic fermentation beverage and the probiotic food.Simplicity in preparation technology is achieved, and as a cereal food, the Lens culinaris is capable of resisting oxidation, lowering blood pressure and preventing cardiovascular and cerebrovascular diseases to different extents through lactobacillus fermentation or without fermentation, and can regulate intestinal tracts, thus, the probiotic-fermented beverage and the solid-fermented food can meet requirements of different consumer groups.
Description
Invention field:
The invention belongs to field of functional food, be utilize multiple probiotic bacteria to mix bacterium solution state, a kind of probiotic bacteria liquid fermentation beverage of solid fermentation Radix Crotalariae szemoensis acquisition and a kind of probiotic food.
Background technology:
In recent years, researching and developing new functional food and increasingly come into one's own, this comes from people to the concern of the health diets such as disease preventing and treating can be helped day by day to rise. The intestinal microbial population of human body can be produced actively impact by probiotic bacteria and prebiotics, makes the focus of research.
Probiotic bacteria is the active microorganism that a class is useful to host, is colonizated in host intestine, reproductive system, can produce definite health efficacy thus improving host's microecological balance, playing the active beneficial microorganism general name of beneficial effect. Common probiotic bacteria mainly includes lactobacillus class, bacillus bifidus class and some bacillus cereus, yeast etc. When probiotic bacteria balance in host, host would be at health status, but internal flora disequilibrium, there is significantly change or during beyond regime values in ratio when between strain, then diarrhoea, allergy, have no appetite, tired, immunity is low etc., and a series of diseases will be following.
Along with vegetarian diet worldwide becomes more and more popular, a lot of countries and regions also have different proportion lactose intolerant patient, and as the milk product of main probiotics fermention goods also containing higher metabolism of lipid and cholesterol, therefore develop the study hotspot that non-breast probiotic composition becomes new. Radix Crotalariae szemoensis, having significantly high nutrition and health care to be worth, wherein in protein, essential amino acids composition is rationally and content of a relatively high (about 25%), fat content relatively low (about 0.7%), carbohydrate content significantly high (60%) containing multivitamin and mineral. Therefore, eat as grain in some countries and regions. Soluble fiber in Radix Crotalariae szemoensis can reduce cholesterol; Ferrum, vitamin B and folate content are also higher, and its Folic Acid is particularly significant to women, it is possible to reduce fetal anomaly rate.
Fermented with Radix Crotalariae szemoensis for substrate, the special flavor material of pleasant can be produced, and select suitable combination bacterial strain mixed fungus fermentation, utilize the synergic fermentation effect between bacterial strain, can improving productivity, promote growth and breeding each other, the product enzyme of combination bacterial strain, product acid and product nutrient substance and the decomposition Utilization ability etc. to substrate to be significantly better than single bacterium and ferment. Combination bacterial strain mixed fungus fermentation also has the beneficial products (GABA (γ-aminobutyric acid), phenols, ACEI (angiotensin converting enzyme inhibitor) etc.) that acquisition pure-blood ferment cannot obtain, and saving of labor is energy-conservation.Therefore, it can, by liquid and two kinds of techniques of solid fermentation, Radix Crotalariae szemoensis is carried out single bacterium fermentation of single bacterial strain and the mixed fungus fermentation of combination bacterial strain, produce probiotic bacteria functional food.
Summary of the invention:
The invention belongs to field of functional food, be utilize multiple probiotic bacteria mixed bacterium that Radix Crotalariae szemoensis carries out fermentation to obtain a kind of probiotics fermention food. Below all for Lactobacillus plantarum and animal bifidobacteria two strain bacterium, carry out the concrete introduction of the present invention.
To achieve these goals, the present invention provides a kind of liquid fermentation technical scheme: a kind of Radix Crotalariae szemoensis probiotic bacteria liquid fermentation beverage, it is as fermentation substrate using Radix Crotalariae szemoensis serosity raw material, adopt Lactobacillus plantarum and animal bifidobacteria after mixed culture liquid state fermentation, through emulsifying, add stabilizer, acid adjustment, perfumery, homogenizing, sterilization after preparation and obtain.
Described Radix Crotalariae szemoensis serosity is to prepare through making beating homogenizing after Radix Crotalariae szemoensis and water being mixed in proportion;
Described Lactobacillus plantarum viable count in fermentation liquid is no less than 1.8 × 108CFU/mL;
Described animal bifidobacteria viable count in fermentation liquid is no less than 1.0 × 108CFU/mL;
Preferably, described Lactobacillus plantarum is Lactobacillus plantarum (Lactobacillusplantarum) TK9, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on the 17th in December in 2015, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postcode 100101, deposit number is CGMCCNo.11891;
Preferably, described animal bifidobacteria is animal bifidobacteria (Bifidobacteriumanimalis) 937, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on the 17th in December in 2015, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, postcode 100101, deposit number is CGMCCNo.11892.
Described Radix Crotalariae szemoensis probiotic bacteria liquid fermentation beverage, is prepared by following method:
(1) actication of culture
Lactobacillus plantarum and animal bifidobacteria glycerol pipe are inoculated in activation medium by 1% respectively and activate, pH7.2-7.4, control temperature and are 34-38 DEG C; After activating 0～20h respectively, carry out re-activation by the bacterium amount that connects of 10% (v/w) activation medium of again transferring, after reactivation 16-24h secondary seed solution;
Described activation medium preparation method is as follows: after being cleaned by Radix Crotalariae szemoensis, namely obtains activation medium after adding 1-10g Radix Crotalariae szemoensis making beating homogenizing in every 100mL water;
(2) mixed fungus fermentation
It is 1 × 10 by Lactobacillus plantarum by connecing bacterium amount7-5×108CFU/mL is inoculated in fermentation medium, and inoculate animal bifidobacteria, Lactobacillus plantarum and animal bifidobacteria to connect bacterium amount ratio be 1-10:1-10 simultaneously, mixed-strains liquid fermentation 12～48h, and temperature is 34-38 DEG C;
Preferably, Lactobacillus plantarum connects bacterium amount is 5 × 108It is 1:10 that CFU/mL, Lactobacillus plantarum and animal bifidobacteria connect bacterium amount ratio;
Preferably, mixed-strains liquid fermentation 28h;
Described fermentation medium preparation method is consistent with activation medium.
(3) fermentation liquor treatment
After fermentation ends through emulsifying, add stabilizer, acid adjustment, perfumery, homogenizing, sterilization after obtain Radix Crotalariae szemoensis fermentation liquid be Radix Crotalariae szemoensis probiotic bacteria liquid fermentation beverage;
Preferably, stabilizer is composed as follows with fermentation liquid mass volume ratio: sodium carboxymethyl cellulose 0.9%, xanthan gum 0.4%, emulsifying agent 0.9%;
Preferably, adding after fermentation ends with fermentation liquid mass volume ratio is the sucrose of 8%;
Preferably, processing condition is that homogenizing temperature 60 DEG C-70 DEG C, pressure 30MPA, homogenization cycles are for once;
Preferably, sterilization conditions is 90 DEG C, 10min.
To achieve these goals, a kind of solid-state fermentation technology scheme provided by the invention: a kind of Radix Crotalariae szemoensis probiotic bacteria solid fermentation food, it is as fermentation substrate using Radix Crotalariae szemoensis raw material, adopts Lactobacillus plantarum and animal bifidobacteria to prepare after solid-state mixed fungus fermentation and obtain;
Described Lactobacillus plantarum viable count in solid medium is no less than 2.1 × 108CFU/g;
Described animal bifidobacteria in solid medium viable count no less than 9.0 × 107CFU/g;
Described Radix Crotalariae szemoensis probiotic bacteria solid fermentation food, is prepared by following method:
The actication of culture mode of solid fermentation is consistent with liquid fermentation;
Further solid fermentation is carried out as follows:
Secondary seed solution is accessed in solid-state fermentation culture medium so that in every gram of solid-state fermentation culture medium, Lactobacillus plantarum connects bacterium amount is 1 × 107-5×108CFU/g, Lactobacillus plantarum and animal bifidobacteria are inoculated in solid medium, mixed culture solid state fermentation 12～48h in the inoculum concentration of the ratio of 1-10:1-10 simultaneously, and temperature is natural, amounts to incubation time 28-92h; After-40 DEG C of pre-freezes 6 hours, put into vacuum freeze drier lyophilizing and be about 16-28h, finally pulverize, sieve, obtain Radix Crotalariae szemoensis solid fermentation food.
Preferably, described Lactobacillus plantarum is Lactobacillus plantarum (Lactobacillusplantarum) TK9;
Preferably, described animal bifidobacteria is animal bifidobacteria (Bifidobacteriumanimalis) 937;
Preferably, Lactobacillus plantarum connects bacterium amount is 1 × 108CFU/g, Lactobacillus plantarum and animal bifidobacteria are inoculated in the ratio of 1:5;
Described solid-state fermentation culture medium preparation method is as follows: after being cleaned by 10g Radix Crotalariae szemoensis, put in 15mL distilled water, soaks 12h at 4 DEG C, and making beating homogenizing obtains Radix Crotalariae szemoensis slurry-mud, adds additive;
Described additive is at least one in calcium carbonate, sucrose, yeast powder, glucose, Pidolidone;
Further, the interpolation concentration of described various additive is respectively as follows: every 25g Radix Crotalariae szemoensis slurry-mud interpolation 0-0.5g;
Preferably, described solid-state fermentation culture medium is: Radix Crotalariae szemoensis 10g, distilled water 15mL, calcium carbonate 0.25g, yeast powder 0.5g, glucose 0.25g, Pidolidone 0.25g;
Preferably, Radix Crotalariae szemoensis solid fermentation food adds protective agent;
Further, described protective agent mass volume ratio is composed as follows: defatted milk powder 5-15%, sodium glutamate 0.5-1.5%, glycerol 2-4%, trehalose 6-8%, dextrin 1-3%, and all the other are water;
Further, described protective agent mass volume ratio is composed as follows: defatted milk powder 10%, sodium glutamate 1%, glycerol 3%, trehalose 8%, dextrin 3%, and all the other are water;
Further, Radix Crotalariae szemoensis solid fermentation products and protectant mass volume ratio are 1:5, and namely 100mL protective agent adds 20g Radix Crotalariae szemoensis solid fermentation food.
Beneficial effect:
In this probiotic bacteria liquid fermentation beverage and solid fermentation food, Lactobacillus plantarum and the highest viable count of animal bifidobacteria are respectively up to 6.21 × 109CFU/mL、4.2×109CFU/g (Lactobacillus plantarum TK9) and 1.3 × 109CFU/mL、1.47×109CFU/g (animal bifidobacteria 937), and rich in GABA (2.605mg/g, 8.132mg/g), total phenols content (7.48mgGAE/g, 28.7mgGAE/g), free amine group content (0.375mmolLeu/g, 1.41mmolLeu/g), polyphenoils content (0.075mmolTE/g, 0.283mmolTE/g), ACEI (22%, 77.9%) prebiotic substance such as, thus Radix Crotalariae szemoensis is as a kind of cereal foods, through lactic acid bacteria fermentation (azymic), there is antioxidation in various degree, blood pressure lowering and the preventive effect to cardiovascular and cerebrovascular disease, conditioning intestinal, preparation technology is simple.Therefore this probiotic fermentation beverage, solid fermentation food and viable bacteria powder preparation are all by the demand of the satisfied different consumer groups. (GAE is gallic acid titer).
And the present invention uses Lactobacillus plantarum and animal bifidobacteria mixed fungus fermentation, effect is better than single bacterium fermentation sum, it is known that Lactobacillus plantarum and in animal bifidobacteria mixed fungus fermentation process, the two is mutually promoted, and creates unforeseeable effect.
Accompanying drawing explanation
Fig. 1: the mixed culture solid state fermentation culture medium additive impact on probiotic bacteria mixed culture solid state fermentation Radix Crotalariae szemoensis food viable count
Wherein, the impact (B) of viable count is the glucose impact on viable count for calcium carbonate by (A)
(C) it is the Pidolidone impact on viable count for yeast powder on the impact (D) of viable count
Fig. 2: the mixed fungus fermentation incubation time impact on probiotic bacteria mixed-strains liquid fermentation Radix Crotalariae szemoensis beverage viable count
Fig. 3: the mixed fungus fermentation incubation time impact on probiotic bacteria mixed culture solid state fermentation Radix Crotalariae szemoensis food viable count
Fig. 4: the single stable agent impact on probiotic bacteria mixed-strains liquid fermentation Radix Crotalariae szemoensis beverage stability
Fig. 5: the sucrose concentration impact on probiotic bacteria mixed-strains liquid fermentation Radix Crotalariae szemoensis beverage sugar-acid ratio
Fig. 6: the processing condition impact on probiotic bacteria mixed-strains liquid fermentation Radix Crotalariae szemoensis beverage stability
Fig. 7: process chart
Specific embodiments
Below in conjunction with form and specific embodiment, the invention will be further described, but the present invention is not limited to specific examples below.
(strain inoculum concentration selects with inoculative proportion, and fermentation parameter selects for embodiment 1, probiotic bacteria liquid fermentation researchonthe technology; Active component changes)
(1) strain inoculum concentration selects with inoculative proportion
Lactobacillus plantarum TK9 and animal bifidobacteria 937 glycerol pipe are inoculated in Radix Crotalariae szemoensis liquid culture medium by 1% respectively and once activate, all activate 20h, then all transfer in re-activation culture medium (with an activation medium) by the bacterium amount that connects of 10% (v/w), re-activation 24h, inoculates Lactobacillus plantarum (L.plantarumTK9) in Radix Crotalariae szemoensis liquid fermentation culture medium simultaneously and animal bifidobacteria 937 (B.animalis937) seed liquor carries out mixed fungus fermentation.
Strain inoculum concentration and inoculative proportion are shown in table 1 below, 2, and at pH7.2-7.4, cultivation temperature 37 ± 1 DEG C, mixed fungus fermentation 24h. Do the comparison of single bacterium fermentation Radix Crotalariae szemoensis liquid fermentation culture medium of Lactobacillus plantarum TK9 and animal bifidobacteria 937 simultaneously respectively, the number of live bacteria of probiotics in all culture medium is detected.
Result: inoculum concentration (see table 1 below, 2) best in liquid fermentation beverage is 5 × 10 for Lactobacillus plantarum TK98It is 1:10 that CFU/mL, Lactobacillus plantarum TK9 and animal bifidobacteria 937 access ratio.
It is 1 × 10 that table 1 Lactobacillus plantarum TK9 connects bacterium amount7-5×108CFU/mL, Lactobacillus plantarum TK9 and animal bifidobacteria 937 connect bacterium ratio when being 1-10:1-10, the viable count of thing lactobacillus TK9.
| TK9 meets bacterium amount CFU/mL | The mono-bacterium comparison of TK9 | 1:1 | 1:5 | 1:10 | 5:1 | 10:1 |
| 1×107 | 2.1×108 | 2.5×108 | 4.5×108 | 1.03×109 | 3.2×108 | 4.6×108 |
| 5×107 | 8.75×108 | 1.25×109 | 1.8×109 | 3.1×109 | 6.5×108 | 1.41×109 |
| 1×108 | 8.95×108 | 1.9×109 | 2.7×109 | 4.17×109 | 9.1×108 | 1.82×109 |
| 5×108 | 9.7×108 | 2.7×109 | 3.3×109 | 5.21×109 | 1.1×109 | 1.93×109 |
It is 1 × 10 that table 2 Lactobacillus plantarum TK9 connects bacterium amount7-5×108CFU/mL, Lactobacillus plantarum TK9 and animal bifidobacteria 937 connect bacterium ratio when being 1-10:1-10, the viable count of animal bifidobacteria 937.
(2) fermentation parameter selects
1), the determination of incubation time
When inoculum concentration optimizes, by 1%, Lactobacillus plantarum TK9 and animal bifidobacteria 937 glycerol pipe are inoculated in Radix Crotalariae szemoensis liquid culture medium respectively cultivate once to activate and all activate 20h, all transfer in Radix Crotalariae szemoensis powder re-activation culture medium by the bacterium amount that connects of 10% (v/w), re-activation 24h, inoculates Lactobacillus plantarum (L.plantarumTK9) and animal bifidobacteria 937 (B.animalis937) seed liquor mixed fungus fermentation in Radix Crotalariae szemoensis liquid fermentation culture medium simultaneously.
Making the Lactobacillus plantarum (L.plantarum) accessed in liquid fermentation culture medium is 5 × 108CFU/mL, the access ratio of Lactobacillus plantarum (L.plantarumTK9) and animal bifidobacteria 937 (B.animalis937) is 1:10, pH7.2-7.4, cultivation temperature 37 ± 1 DEG C, mixed fungus fermentation 4,10,16,22,28,34,40h, respectively the number of live bacteria of probiotics in culture medium is detected.
Result: mixed-strains liquid fermentation the best incubation time is 28h, total incubation time is 72h (see accompanying drawing 2). The total viable count recorded is 7.51 × 109CFU/mL, wherein Lactobacillus plantarum (L.plantarumTK9) viable count is 6.21 × 109CFU/mL, animal bifidobacteria 937 (B.animalis937) viable count are 1.3 × 109CFU/mL。
2), the determination of emulsifying agent and stabilizer in Radix Crotalariae szemoensis liquid fermentation beverage
The technological process of production of Radix Crotalariae szemoensis fermented beverage is as shown in Figure 7;
The stabilizers such as sodium carboxymethyl cellulose, sodium alginate, carrageenan, pectin, xanthan gum are made an addition in Radix Crotalariae szemoensis fermented beverage by variable concentrations, at 60 DEG C-70 DEG C, when 30MPa, homogenizing is once, after 90 DEG C sterilize 15min, with centrifugation rate for index, investigate the impact on Radix Crotalariae szemoensis fermented beverage stability of the different stabilizers, and then select preferably stabilizer.
Result: as seen from the figure, when sodium carboxymethyl cellulose and xanthan gum addition are respectively when 0.2%-0.4%, the centrifugation rate of Radix Crotalariae szemoensis fermented beverage reduces notable. When sodium carboxymethyl cellulose and xanthan gum addition are respectively when 0.4%-0.6%, the centrifugation rate of Radix Crotalariae szemoensis fermented beverage is almost constant. When sodium carboxymethyl cellulose and xanthan gum addition are 0.6%, its sedimentation rate is reduced to 2.2%, 2.3% respectively. And during identical addition, sodium alginate, carrageenan, pectin interpolation can not reach and add sodium carboxymethyl cellulose and the suitable effect of xanthan gum. (see accompanying drawing 4).
Sodium carboxymethyl cellulose (CMC-Na), xanthan gum and emulsifying agent are carried out orthogonal test as three factors, with coefficient of stability for index, empirical factor level such as following table:
Result: can be obtained by following orthogonal experiments, the impact order of Radix Crotalariae szemoensis fermented beverage stability is A > B > C by it, and the optimum proportioning of complex stabilizer is A3B2C3, i.e. CMC-Na0.9%, xanthan gum 0.4%, emulsifying agent 0.9%. Wherein CMC-Na is notable to Radix Crotalariae szemoensis fermented beverage stability influence.
3), sugar-acid ratio is determined
Sucrose is added in Radix Crotalariae szemoensis fermented beverage by the Concentraton gradient of 4%, 6%, 8%, 10%, with subjective appreciation for inspection target.
Result: when sucrose addition is 8% (see accompanying drawing 5), Radix Crotalariae szemoensis fermented beverage has its distinctive fresh scent, and tissue is fine and smooth uniformly, in lilac, substantially precipitates out without clear liquid, generates without precipitation, and sour-sweet suitable, now sensory evaluation is preferred.
4), processing condition is determined
By above-mentioned fermented beverage at 60 DEG C-70 DEG C, the barometric gradient by certain: 20,30,40MPA homogenizing once or twice, with beverage sedimentation rate for index.
Result: Radix Crotalariae szemoensis fermented beverage homogenizing temperature 60 DEG C-70 DEG C, pressure 30MPA, homogenization cycles be once when, beverage stability preferably (see accompanying drawing 6).
5), sterilization conditions is determined
By above-mentioned fermented beverage when sterilization temperature respectively 85,90,95,100 DEG C, each temperature carry out respectively sterilization 10,15,20min, with total plate count, coliform, mycete and yeast, pathogenic bacterium for index, detect with reference to GB4789.2.3 1.
Result: best sterilization conditions is 90 DEG C, 10min. Under this sterilization conditions, the microbiological indicator of Radix Crotalariae szemoensis fermented beverage is as follows
(3) culture medium active component change (GABA, total phenols, free ammonical nitrogen, polyphenoils, ACEI) before and after fermentation
Fermentation condition:
Lactobacillus plantarum and animal bifidobacteria glycerol pipe are inoculated in activation medium by 1% respectively and activate, pH7.3, and controlling temperature is 37 DEG C; Respectively after activation 10h, carry out re-activation by the bacterium amount that connects of 10% (v/w) activation medium of again transferring, after reactivation 20h secondary seed solution;
Described activation medium preparation method is as follows: after being cleaned by Radix Crotalariae szemoensis, namely obtains activation medium after adding 5g Radix Crotalariae szemoensis making beating homogenizing in every 100mL water;
It is 5 × 10 by Lactobacillus plantarum by connecing bacterium amount8CFU/mL is inoculated in fermentation medium, and inoculate animal bifidobacteria, Lactobacillus plantarum and animal bifidobacteria to connect bacterium amount ratio be 1:10 simultaneously, mixed-strains liquid fermentation 28h, and temperature is 37 DEG C;
Described fermentation medium preparation method is consistent with activation medium.
(3) fermentation liquor treatment
After fermentation ends through emulsifying, add stabilizer, acid adjustment, perfumery, homogenizing, sterilization after obtain Radix Crotalariae szemoensis fermentation liquid be Radix Crotalariae szemoensis probiotic bacteria liquid fermentation beverage;
Stabilizer is composed as follows with fermentation liquid mass volume ratio: sodium carboxymethyl cellulose 0.9%, xanthan gum 0.4%, emulsifying agent 0.9%;
Adding after fermentation ends with fermentation liquid mass volume ratio is the sucrose of 8%;
Processing condition is that homogenizing temperature 65 DEG C, pressure 30MPA, homogenization cycles are for once;
Sterilization conditions is 90 DEG C, 10min.
Utilize Syrups by HPLC GABA content; Article (Rozan with reference to Rozan et al., Kuo, andLambein (2000) .Freeaminoacidspresentincommerciallyavailableseedlingsso ldforhumanconsumption.Apotentialhazardforconsumers.Journ alofAgricultureandFoodChemistry, 48,716 723.);
Utilize the total phenols content of forint phenol reaction assay; Article (Adler-Nissen with reference to Adler-Nissen et al., J. (1979) .Determinationofthedegreeofhydrolysisoffoodproteinhydrol ysatesbytrinitrobenzenesulfonicacid.JournalofAgricultura landFoodChemistry, 27,1256 1262.);
2,4,6-trinitro-benzene-sulfonic acid and microplate reader is utilized to measure free amine group content;
Oxygen index method (ORAC) of living is utilized to measure polyphenoils content;
ACEI content assaying method is with reference to Hyun and Shin et al. article (Hyun, C.K., &Shin, H.K. (2000) .Utilizationofbovinebloodplasmaproteinsfortheproductiono fangotensinIconvertingenzymeinhibitorypeptides.ProcessBi ochemistry, 36,65 71.).
Liquid fermentation result such as following table:
Note: single same mixed fungus fermentation of bacterium fermentation process, it is only necessary to select single culture corresponding in mixed fungus fermentation and the inoculum concentration of corresponding single culture.
By upper form it can be seen that in Radix Crotalariae szemoensis fermented beverage GABA content, total phenols, free amine group, polyphenoils and ACEI content all increase significantly. Particularly, after Lactobacillus plantarum (CGMCC1.6971) and animal bifidobacteria (CGMCC1.3003) mixed fungus fermentation, GABA content after GABA content and Lactobacillus plantarum TK9 and animal bifidobacteria 937 mixed fungus fermentation is above the summation of single bacterium tunning, and hypertension is had by this composition to be alleviated and effect for the treatment of;The Fermentation Data of free amine group shows that the effect of mixed fungus fermentation is better than single bacterium fermentation sum equally.
Embodiment 2, solid-state fermentation process (strain inoculum concentration and inoculative proportion select, fermentation parameter selects, active component changes)
(1) strain inoculum concentration selects with inoculative proportion
Lactobacillus plantarum TK9 and animal bifidobacteria 937 glycerol pipe are inoculated in Radix Crotalariae szemoensis liquid culture medium by 1% respectively and once activate, all activate 20h, then all transfer in re-activation culture medium (with an activation medium) by the bacterium amount that connects of 10% (v/w), re-activation 24h, inoculates Lactobacillus plantarum (L.plantarumTK9) in Radix Crotalariae szemoensis solid-state fermentation culture medium simultaneously and animal bifidobacteria 937 (B.animalis937) seed liquor carries out mixed fungus fermentation.
Strain inoculum concentration and inoculative proportion are shown in table 3 below, 4, and at pH7.2-7.4, cultivation temperature 37 ± 1 DEG C, after mixed fungus fermentation 24h. Do the comparison of single bacterium fermentation Radix Crotalariae szemoensis solid-state fermentation culture medium of Lactobacillus plantarum TK9 and animal bifidobacteria 937 simultaneously respectively, the number of live bacteria of probiotics in all culture medium is detected.
Result shows: inoculum concentration best in solid fermentation beverage is Lactobacillus plantarum TK9 is 1 × 108It is 1:5 (see accompanying drawing 2) that CFU/g, Lactobacillus plantarum TK9 and animal bifidobacteria 937 access ratio.
It is 1 × 10 that table 3 Lactobacillus plantarum TK9 connects bacterium amount7-5×108CFU/g, Lactobacillus plantarum TK9 and animal bifidobacteria 937 connect bacterium ratio when being 1-10:1-10, the viable count of thing lactobacillus TK9.
| TK9 meets bacterium amount CFU/g | The mono-bacterium comparison of TK9 | 1:1 | 1:5 | 1:10 | 5:1 | 10:1 |
| 1×107 | 2.7×108 | 2.9×108 | 5.2×108 | 1.47×109 | 3.9×108 | 5.7×108 |
| 5×107 | 9.2×108 | 9.2×108 | 2.21×109 | 3.4×109 | 8.1×108 | 9.95×108 |
| 1×108 | 2.17×109 | 2.17×109 | 3.9×109 | 2.74×109 | 1.02×109 | 3.1×109 |
| 5×108 | 8.75×108 | 1.02×109 | 2.1×109 | 3.15×109 | 1.1×109 | 1.47×109 |
It is 1 × 10 that table 4 Lactobacillus plantarum TK9 connects bacterium amount7-5×108CFU/g, Lactobacillus plantarum TK9 and animal bifidobacteria 937 connect bacterium ratio when being 1-10:1-10, the viable count of animal bifidobacteria 937.
(2) fermentation parameter selects
1), the determination of fermentation culture based additive
When inoculum concentration optimizes, by 1%, Lactobacillus plantarum TK9 and animal bifidobacteria 937 glycerol pipe are inoculated in Radix Crotalariae szemoensis liquid culture medium respectively cultivate once to activate and all activate 20h, all transfer in Radix Crotalariae szemoensis powder re-activation culture medium by the bacterium amount that connects of 10% (v/w), re-activation 24h.
Fermentation medium: add variable concentrations gradient (0,0.25,0.5g/25mL) calcium carbonate, yeast powder, glucose, Pidolidone in Radix Crotalariae szemoensis slurry respectively.
Radix Crotalariae szemoensis solid-state fermentation culture medium is inoculated Lactobacillus plantarum (L.plantarumTK9) and animal bifidobacteria 937 (B.animalis937) seed liquor mixed fungus fermentation simultaneously. Making the Lactobacillus plantarum (L.plantarum) accessed in solid-state fermentation culture medium is 1 × 108CFU/g, the access ratio of Lactobacillus plantarum (L.plantarumTK9) and animal bifidobacteria 937 (B.animalis937) is 1:5, pH7.2-7.4, cultivation temperature 37 ± 1 DEG C, mixed fungus fermentation 24h, detects the number of live bacteria of probiotics in culture medium respectively.
Result: mixed culture solid state fermentation culture medium optimum formula is: Radix Crotalariae szemoensis 10g, distilled water 15mL, calcium carbonate 0.25g, yeast powder 0.5g, glucose 0.25g, Pidolidone 0.25g (see accompanying drawing 1).
When calcium carbonate addition is 0.25g (/ 25g), the total viable count recorded is 4.395 × 109CFU/g, wherein Lactobacillus plantarum (L.plantarumTK9) viable count is 3.7 × 109CFU/g, animal bifidobacteria 937 (B.animalis937) viable count are 6.95 × 108CFU/g; When yeast powder addition is 0.5g (/ 25g), the total viable count recorded is 4.82 × 109CFU/g, wherein Lactobacillus plantarum (L.plantarumTK9) viable count is 3.9 × 109CFU/g, animal bifidobacteria 937 (B.animalis937) viable count are 9.2 × 108CFU/g;When glucose addition is 0.25g (/ 25g), the total viable count recorded is 3.825 × 109CFU/g, wherein Lactobacillus plantarum (L.plantarumTK9) viable count is 3.1 × 109CFU/g, animal bifidobacteria 937 (B.animalis937) viable count are 7.25 × 108CFU/g; When Pidolidone addition is 0.25g (/ 25g), the total viable count recorded is 4.89 × 109CFU/g, wherein Lactobacillus plantarum (L.plantarumTK9) viable count is 3.8 × 109CFU/g, animal bifidobacteria 937 (B.animalis937) viable count are 1.09 × 109CFU/g。
2), the determination of incubation time
When inoculum concentration optimizes, by 1%, Lactobacillus plantarum TK9 and animal bifidobacteria 937 glycerol pipe are inoculated in Radix Crotalariae szemoensis liquid culture medium respectively cultivate once to activate and all activate 20h, all transfer activation 24h in re-activation culture medium by the bacterium amount that connects of 10% (v/w), Radix Crotalariae szemoensis solid-state fermentation culture medium is inoculated Lactobacillus plantarum (L.plantarumTK9) and animal bifidobacteria 937 (B.animalis937) seed liquor mixed fungus fermentation simultaneously.
Making the Lactobacillus plantarum (L.plantarum) accessed in solid-state fermentation culture medium is 1 × 108CFU/g, the access ratio of Lactobacillus plantarum (L.plantarumTK9) and animal bifidobacteria 937 (B.animalis937) is 1:5, pH7.2-7.4, cultivation temperature 37 ± 1 DEG C, mixed fungus fermentation 4,10,16,22,28,34,40h, respectively the number of live bacteria of probiotics in culture medium is detected.
Result: mixed culture solid state fermentation the best incubation time is 34h, total incubation time is 78h (see accompanying drawing 3). The total viable count recorded is 5.67 × 109CFU/g, wherein Lactobacillus plantarum (L.plantarumTK9) viable count is 4.2 × 109CFU/g, animal bifidobacteria 937 (B.animalis937) viable count are 1.47 × 109CFU/g。
3), the protectant selection of mixed culture solid state fermentation product vacuum lyophilization
Defatted milk powder, sodium glutamate, glycerol, trehalose, dextrin are added into by different concentration in the culture medium fermented respectively, culture medium and protective agent ratio 1:5 (namely 100ml protective agent adds 20g culture medium); After stirring at-40 DEG C pre-freeze 6h ,-60 DEG C carry out vacuum lyophilization, and vacuum is 4pa, vacuum lyophilization 20h, then pulverizes, and sieves. In lyophilized powder, viable count is inspection target, and orthogonal experiment factor level is shown in following table:
Result: above-mentioned freeze drying protectant Orthogonal experiment results is carried out range analysis, show that the order to Radix Crotalariae szemoensis solid fermentation food frozen-dried protective effect is A > B > E > D > C, the optimum formula of freeze drying protectant is than for A2B2E3D3C3, i.e. defatted milk powder 10%, sodium glutamate 1%, dextrin 3%, trehalose 8%, glycerol 3%.
(3) active component change (GABA, total phenols, free ammonical nitrogen, polyphenoils, ACEI) before and after fermentation
Fermentation condition:
Lactobacillus plantarum and animal bifidobacteria glycerol pipe are inoculated in activation medium by 1% respectively and activate, pH7.3, and controlling temperature is 35 DEG C; Respectively after activation 10h, carry out re-activation by the bacterium amount that connects of 10% (v/w) activation medium of again transferring, after reactivation 20h secondary seed solution;
Described activation medium preparation method is as follows: after being cleaned by Radix Crotalariae szemoensis, namely obtains activation medium after adding 5g Radix Crotalariae szemoensis making beating homogenizing in every 100mL water;
Further solid fermentation is carried out as follows:
Secondary seed solution is accessed in solid-state fermentation culture medium so that in every gram of solid medium, Lactobacillus plantarum connects bacterium amount is 1 × 108CFU/g, Lactobacillus plantarum and animal bifidobacteria are inoculated in solid medium, mixed culture solid state fermentation 34h in the inoculum concentration of the ratio of 1:5 simultaneously, and temperature is natural, amounts to incubation time 64h;After-40 DEG C of pre-freezes 6 hours, put into vacuum freeze drier lyophilizing and be about 20h, finally pulverize, sieve, obtain Radix Crotalariae szemoensis solid fermentation food.
Described solid-state fermentation culture medium is: Radix Crotalariae szemoensis 10g, distilled water 15mL, calcium carbonate 0.25g, yeast powder 0.5g, glucose 0.25g, Pidolidone 0.25g;
The fresh bent material of Radix Crotalariae szemoensis solid fermentation adds protective agent;
Described protective agent mass volume ratio is composed as follows: defatted milk powder 10%, sodium glutamate 1%, glycerol 3%, trehalose 8%, dextrin 3%, and all the other are water.
The same liquid fermentation of parametric measurement method.
Solid fermentation result such as following table:
Note: single same mixed fungus fermentation of bacterium fermentation process, it is only necessary to select single culture corresponding in mixed fungus fermentation and the inoculum concentration of corresponding single culture.
From upper form; in Radix Crotalariae szemoensis solid fermentation food, GABA content, total phenols, free amine group, polyphenoils and ACEI content all increase significantly, particularly TK9, the content 1.49mmolLeu/g of free amine group is higher than the 0.63mmolLeu/g sum of the single bacterium fermentation in 0.77mmolLeu/g and 937 of TK9 mono-bacterium fermentation after 937 mixed fungus fermentations.
Claims (10)
1. a Radix Crotalariae szemoensis fermented food, it is characterised in that described Radix Crotalariae szemoensis fermented food is with Radix Crotalariae szemoensis for fermentation substrate, adopts Lactobacillus plantarum and animal bifidobacteria to prepare after liquid or solid-state mixed fungus fermentation and obtain.
2. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 1, it is characterised in that described Radix Crotalariae szemoensis fermented food is prepared with liquid fermentation method, specific as follows:
(1) actication of culture
Lactobacillus plantarum and animal bifidobacteria glycerol pipe are inoculated in activation medium by 1% respectively and activate, pH7.2-7.4, control temperature and are 34-38 DEG C; After activating 0～20h respectively, carry out re-activation by the bacterium amount that connects of 10% activation medium of again transferring, after reactivation 16-24h secondary seed solution;
Described activation medium preparation method is as follows: after being cleaned by Radix Crotalariae szemoensis, namely obtains activation medium after adding 1-10g Radix Crotalariae szemoensis making beating homogenizing in every 100mL water;
(2) mixed fungus fermentation
It is 1 × 10 by Lactobacillus plantarum by connecing bacterium amount7-5×108CFU/mL is inoculated in fermentation medium, and inoculate animal bifidobacteria, Lactobacillus plantarum and animal bifidobacteria to connect bacterium amount ratio be 1-10:1-10 simultaneously, mixed-strains liquid fermentation time 12～48h, and temperature is 34-38 DEG C;
Described fermentation medium preparation method is consistent with activation medium;
Described Lactobacillus plantarum viable count in fermentation liquid is no less than 1.8 × 108CFU/mL; Described animal bifidobacteria viable count in fermentation liquid is no less than 1.0 × 108CFU/mL;
(3) fermentation liquor treatment
Fermentation ends after fermentation liquid through emulsifying, add stabilizer, acid adjustment, perfumery, homogenizing, sterilization after the fresh bent material of Radix Crotalariae szemoensis liquid fermentation that obtains be Radix Crotalariae szemoensis probiotic bacteria liquid fermentation beverage.
3. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 2, it is characterised in that it is 5 × 10 that Lactobacillus plantarum described in mixed fungus fermentation connects bacterium amount8It is 1:10 that CFU/mL, Lactobacillus plantarum and animal bifidobacteria connect bacterium amount ratio; Described mixed-strains liquid fermentation is 28h.
4. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 2, it is characterised in that
Described stabilizer is by composed as follows with fermentation liquid mass volume ratio: sodium carboxymethyl cellulose 0.9%, xanthan gum 0.4%, emulsifying agent 0.9%;
Described acid adjustment is that after fermentation ends, interpolation and fermentation liquid mass volume ratio are the sucrose of 8%;
The condition of described homogenizing is that homogenizing temperature 60 DEG C-70 DEG C, pressure 30MPA, homogenization cycles are for once;
The condition of described sterilization is 90 DEG C, 10min.
5. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 1, it is characterised in that described Radix Crotalariae szemoensis fermented food is prepared with solid state fermentation, specific as follows:
(1) actication of culture
Lactobacillus plantarum and animal bifidobacteria glycerol pipe are inoculated in activation medium by 1% respectively and activate, pH7.2-7.4, control temperature and are 34-38 DEG C; After activating 0～20h respectively, carry out re-activation by the bacterium amount that connects of 10% activation medium of again transferring, after reactivation 16-24h secondary seed solution;
Described activation medium preparation method is as follows: after being cleaned by Radix Crotalariae szemoensis, namely obtains activation medium after adding 1-10g Radix Crotalariae szemoensis making beating homogenizing in every 100mL water;
(2) mixed fungus fermentation
Secondary seed solution is accessed in solid-state fermentation culture medium so that in every gram of solid medium, Lactobacillus plantarum connects bacterium amount is 1 × 107-5×108CFU/g, Lactobacillus plantarum and animal bifidobacteria are inoculated in solid medium, mixed culture solid state fermentation 12～48h in the inoculum concentration of the ratio of 1-10:1-10 simultaneously, and temperature is natural, amounts to incubation time 28-92h;
(3) post processing
After having fermented by fermentation substrate after-40 DEG C of pre-freezes 6 hours, put into vacuum freeze drier lyophilizing and be about 16-28h, finally pulverize, sieve, obtain the fresh bent material of Radix Crotalariae szemoensis solid fermentation.
6. a kind of Radix Crotalariae szemoensis fermented food as described in claim 1,2 or 5 any one, it is characterised in that described Lactobacillus plantarum is Lactobacillus plantarum (Lactobacillusplantarum) TK9, and deposit number is CGMCCNo.11891; Described animal bifidobacteria is animal bifidobacteria (Bifidobacteriumanimalis) 937, and deposit number is CGMCCNo.11892.
7. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 5, it is characterised in that it is 1 × 10 that described Lactobacillus plantarum connects bacterium amount8CFU/g, Lactobacillus plantarum and animal bifidobacteria are inoculated in the ratio of 1:5.
8. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 5, it is characterised in that the preparation method of described solid-state fermentation culture medium is as follows: after being cleaned by 10g Radix Crotalariae szemoensis, put in 15mL distilled water, soaking 12h at 4 DEG C, making beating homogeneous preparation becomes Radix Crotalariae szemoensis mud, adds additive;
Described additive is at least one in calcium carbonate, sucrose, yeast powder, glucose, Pidolidone;
The interpolation concentration of described every kind of additive is: every 25g Radix Crotalariae szemoensis slurry-mud adds additive 0-0.5g.
9. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 8, it is characterised in that described solid-state fermentation culture medium is: Radix Crotalariae szemoensis 10g, distilled water 15mL, calcium carbonate 0.25g, yeast powder 0.5g, glucose 0.25g, Pidolidone 0.25g.
10. a kind of Radix Crotalariae szemoensis fermented food as claimed in claim 5; it is characterized in that; the fresh bent material of described Radix Crotalariae szemoensis solid fermentation is added with protective agent; described protective agent mass volume ratio is composed as follows: defatted milk powder 5-15%; sodium glutamate 0.5-1.5%, glycerol 2-4%, trehalose 6-8%, dextrin 1-3%, all the other are water.
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| CN113662139A (en) * | 2021-08-23 | 2021-11-19 | 黑龙江八一农垦大学 | Bean composite fermentation liquor and preparation method thereof |
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