CN102754782A - Preparation method of fermentation type fruit and vegetable powder rich in gamma-aminobutyric acid and lactobacillus plantarum - Google Patents
Preparation method of fermentation type fruit and vegetable powder rich in gamma-aminobutyric acid and lactobacillus plantarum Download PDFInfo
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Abstract
The invention provides a preparation method of fermentation type fruit and vegetable powder rich in gamma-aminoburyric acid and lactobacillus plantarum. According to the preparation method, screened lactobacillus plantarum capable of generating GABA (Gamma-Aminoburyric Acid) is taken as fermentation strains, fruits and vegetables are taken as basic raw materials, a fruit and vegetable fermentation liquor is obtained by utilizing metabolization to control anaerobic digestion and conducting spray drying, the content of GABA and the viable count of bacterium lacticum are high, the GABA is more than 6.9g/L, the viable count is more than 5*109cuf/ml, and the yield of the fruit and vegetable powder is 8%-20%. According to the preparation method, the lactobacillus plantarum is utilized for preparing the fruit and vegetable powder rich in GABA and lactobacillus plantarum, so that not only is the nutrition of the fruit and vegetable powder improved, but also the preparation method is the technological extension of the processing of the fruit and vegetable powder, and the additional value of the fruit and vegetable powder is increased, so that the preparation method has a practical significance and an application prospect.
Description
(1) technical field
The present invention relates to a kind of preparation method who is rich in the fermented type fruits and vegetables powder of GABA (GABA) and Lactobacillus plantarum.
(2) background technology
Because GABA has the important physical function, at aspects such as development functionality cereal article, beverage and health products good application potential is arranged so produce the Lactobacillus plantarum of GABA, these products not only contain GABA, also contain abundant probio.Add to after the soy sauce in the ferment process like document report Lactobacillus plantarum, can produce the functional form soy sauce that is rich in GABA and Lactobacillus plantarum.Utilize cheese production to make that the content of GABA has reached 383 mg/kg in the cheese.Park filters out Lactobacillus brevis fermentation and obtains GABA output and reach 2.5 g/L from pickles, and uses germinated soybean extract and milk to be fermented material, and adding mixing thalline carries out the ferment making sour milk, obtains that GABA content is 424.67 μ g/g in the sour milk.
Work for the exploitation Japanology that is rich in the GABA functional food is more; More existing realized commercialization production, like the Jia Yelong tea of Japan, Ao Lizha company produces is rich in GABA rice germ extracts; The sprouted unpolished rice of Fa Kaier company; Also having ocean spices company is the GABA enriched food " good Braak holder " that primary raw material is produced through lactic fermentation with cow's milk and natural glutamic acid, and excellent flavor is fit to process drinks and food.And China is in the starting stage in this respect, and at present, China Zhejiang benefit ten thousand Bioisystech Co., Ltd successfully produce the GABA food additives that are rich in GABA rice plumule and Anhui Life Co., Ltd. production etc.
The southern fruits and vegetables aboundresources of present China; Cheap, and garden stuff processing industry seriously lags behind the development of plant husbandry, exists every year a large amount of inferior fruits and vegetables and the residue that can't sell really to dredge; Make the fruits and vegetables powder product that is rich in GABA if utilize Lactobacillus plantarum; Not only improve the nutrition of fruits and vegetables powder, and be the technology extension of garden stuff processing, increased an important directions of fruits and vegetables added value.Nowadays GABA and Lactobacillus plantarum are new resource food by Ministry of Health of the People's Republic of China's approval, therefore utilize the fruits and vegetables powder that the Lactobacillus plantarum fermenting process of preparing is rich in GABA to have more realistic meaning and wide prospect.
(3) summary of the invention
The object of the invention provides a kind of production technology that is rich in the fermented type fruits and vegetables powder of GABA (GABA) and Lactobacillus plantarum.
The technical scheme that the present invention adopts is:
A kind of preparation method who is rich in the fermented type fruits and vegetables powder of GABA and Lactobacillus plantarum, said method comprises:
(1) with raw material fruits and vegetables peeling squeezing, add cellulase and pectase (acid pectase) complex enzyme powder and carry out the enzyme processing, centrifugal filtration is got supernatant and is obtained Juice again;
(2) Juice being diluted to sugar content is 2～50g/L, adds yeast extract 20 g/L again, sodium acetate 2 g/L, and sodium glutamate 2～50 g/L, regulating the pH value is 6.6～6.8 (allocating with citric acid, hydrochloric acid or NaOH), obtains fermentation medium; Need to allocate, generally need dilution to make sugar content roughly at 2～50g/L for 2 ~ 10 times, to reduce high sugar content producing the inhibitory action of GABA according to the difference of the sugared content in the Juice;
(3) fermentation medium is after sterilization, and inoculation Lactobacillus plantarum (Lactobacillus plantarum) CCTCC NO:M 2011364 leaves standstill cultivation 80～200h 30～35 ℃ of following anaerobism, obtains zymotic fluid; Bacterial strain need carry out the seed enlarged culture usually before being seeded to fermentation medium; Used seed culture medium is the MRS culture medium; Initial pH value is 6.2～6.4; Condition of culture is that 30～35 ℃ of following anaerobism are cultivated 18～24h, obtains seed liquor and is seeded to fermentation medium with 2 ~ 5% volume ratio inoculum concentrations again; After the fermentation ends, adopt high performance liquid chromatography can more accurately draw the content of the GABA in the fruits and vegetables powder of fermentation back, adopt different dilution factors to coat on the solid fruit-vegetable juice fermentation medium simultaneously and can draw clump count;
(4) zymotic fluid concentrates (generally being concentrated into solid content 30 ~ 40% gets final product), spray-drying, promptly gets said fermented type fruits and vegetables powder.
Said raw material fruits and vegetables are one of following: citrus, apple, grape, strawberry, tomato, watermelon.
Preferably, cellulase and pectase enzyme powder consumption are 1:1 (among the present invention, the enzyme of used cellulose enzyme powder is lived and is that it is 30000U/g that 10000U/g, the enzyme of pectase enzyme powder live, and both mass ratioes are 3:1) with enzyme work in the said step (1).The complex enzyme total addition level is 2 ~ 50,000 U/kg Juices.
Said step has been added glutamic acid or sodium glutamate in (2), and Lactobacillus plantarum CCTCC NO:M 2011364 of the present invention can produce GABA in the culture medium that with glutamic acid or sodium glutamate is substrate.
Raw materials used when being citrus; Because citrus contains a large amount of bitter substances; Main component is an aurantiin, therefore need carry out the enzyme process debitterize in the anaerobic fermentation later stage and handle, to remove wherein bitter substance; Can effectively reduce the bitter taste of zymotic fluid; Be specially: the naringinase (debitterize enzyme) that when step (3) anaerobic fermentation to 60～150 h, adds 500 ~ 1200U/L fermentation medium carries out synchronous debitterize, and said naringinase is obtained through fermentation by aspergillus niger (Aspergillus niger) CCTCC NO:M 206047, and concrete bacterial strain information and fermentation process are referring to CN101089175A.
The bacterial strain that the process that the invention still further relates to is used---Lactobacillus plantarum (Lactobacillus plantarum) WZ011; Be preserved in Chinese typical culture collection center, address: China, Wuhan; Wuhan University; Postcode 430072, preservation date on October 24th, 2011, deposit number CCTCC No:M 2011364.
Said step (4) spray-drying condition is: 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.Products obtained therefrom discharging moisture≤6%, the sealing of cooling back places 4 ℃ of dry place's preservations.
Utilization of the present invention sieved the Lactobacillus plantarum that can produce GABA be fermentation strain; With fruits and vegetables is base stock, and through metabolism control anaerobic fermentation and low temperature drying, the GABA and the living preparation of lactobacillus that obtain in the fruits and vegetables zymotic fluid are counted the content height; GABA can reach 6.9 g/L, and viable count can reach 5 * 10
8Cfu/ml, the yield of fruits and vegetables powder are 8%～20%.Utilize Lactobacillus plantarum to make the fruits and vegetables powder product that is rich in GABA and Lactobacillus plantarum, not only improve the nutrition of fruits and vegetables powder, and be the technology extension of fruits and vegetables powder processing, increase fruits and vegetables powder added value and have realistic meaning and wide prospect.
(4) specific embodiment
Below in conjunction with specific embodiment the present invention is described further, but protection scope of the present invention is not limited in this:
Embodiment 1:
(A) raw material preparation: In November picking citrus as raw materials, peeled juice, citrus pulp obtained crude 50kg, citrus pulp in the crude cellulase and pectinase added mass ratio of 3:1 complexes enzyme powder enzyme treatment, compound enzyme preparation of 120g (cellulase: 10000u / g, acid pectinase: activity 30000u / g, were purchased from Wuxi Xuemei preparations Ltd.), enzyme treatment time was 90min; then centrifugation to remove skins, nuts and other residue was orange juice.The allotment of orange juice dilution: glucose content is 34.5 g/L before the orange juice dilution, dilute 6 times after, sugared content is 5.95g/L.
(2) allotment of orange juice medium component and fermentation procedure
(1) adds yeast extract 20 g/L again to above-mentioned dilution orange juice, sodium acetate 2 g/L, sodium glutamate 15 g/L; Medium pH is transferred to 6.4～6.6, and 100 ℃ of sterilization 20min are cooled to room temperature.
(2) preservation of bacterial classification and cultural method:
1) bacterial classification: Lactobacillus plantarum WZ011 (being CCTCC No:M 2011364);
2) culture medium and condition of culture
Seed culture medium (MRS nutrient solution): peptone 10 g/L, beef extract 10 g/L, yeast extract 5 g/L, glucose 20 g/L, sodium acetate 5 g/L, dibasic ammonium citrate 2 g/L, dipotassium hydrogen phosphate 2 g/L, Tween-80 1 mL/L, MnSO
44H
2O 0.25 g/L, MgSO
47H
2O 0.58 g/L, solvent are water, and pH 6.2～6.4.
Slant medium: with seed culture medium (adding agar 20g/L).
Seed culture: the 50 mL MRS nutrient solutions of packing in the 250 mL triangular flasks, the lactic acid bacteria of picking purifying goes into the MRS nutrient solution, seals with preservative film, leaves standstill in 30 ℃ of anaerobism and cultivates 18 h.
(3) fermentation condition: seed liquor inoculum concentration (v/v) is 4%, and the orange juice fermentation medium leaves standstill 35 ℃ of following anaerobism cultivates 120h, initial pH=6.4～6.6.The naringinase that adds 700 U/L fermentation mediums during anaerobic fermentation to 80 h carries out synchronous debitterize (obtained through fermentation by aspergillus niger (Aspergillus niger) CCTCC NO:M 206047, the naringinase vigor is 20000U/g).
(4) 35 ℃ leave standstill cultivate 120 h after, adopt the GABA content in the liquid chromatography for measuring orange juice culture medium, draw GABA output and reach 6.9 g/L.Adopt coating orange juice solid fermentation medium therapy to record that the Lactobacillus plantarum viable count reaches 5 * 10 in the orange juice zymotic fluid again
8Cfu/ml, the sodium glutamate utilization rate reaches 87.9%.
(3) post processing:
The concentrate of orange juice culture medium is carried out spray-drying processing, and input concentration is about 40%, 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.
Dry back citrus fruit powder gets yield and is: 15%, and moisture≤4%.Viable count reaches 4.3 * 10
9Cfu/g, GABA content are 6.2%.
Embodiment 2:
(A) raw material preparation: In November picking citrus as raw materials, peeled juice, take 50kg sauce added cellulase and pectinase enzyme mass ratio of 3:1 powder (cellulase: 10000u / g , acid pectinase: activity 30000u / g, were purchased from Wuxi Xuemei preparations Ltd.) 120g of the enzyme treatment to obtain a crude orange juice, and then centrifuged to remove the bladder, the nucleolus and other debris, and then wrap with plastic membrane seal placed at 4 ℃ refrigerator cold standby.The allotment of orange juice dilution: glucose content is 56 g/L before the orange juice dilution, dilute 5 times after, sugared content is 10.11g/L.
(2) allotment of orange juice medium component and fermentation procedure
(1) adds yeast extract 20 g/L again to above-mentioned dilution orange juice, sodium acetate 2 g/L, glutamic acid 15 g/L.Medium pH is transferred to 6.4～6.6.100 ℃ of sterilizations are 20min, are cooled to room temperature.
(2) preservation of bacterial classification and cultural method:
1) bacterial classification: Lactobacillus plantarum WZ011
2) culture medium and condition of culture
Seed culture medium (MRS nutrient solution): peptone 10 g/L, beef extract 10 g/L, yeast extract 5 g/L, glucose 20 g/L, sodium acetate 5 g/L, dibasic ammonium citrate 2 g/L, dipotassium hydrogen phosphate 2 g/L, Tween-80 1 mL/L, MnSO
44H
2O 0.25 g/L, MgSO
47H
2O 0.58 g/L, solvent are water, and pH 6.2～6.4.
Slant medium: with seed culture medium (adding agar 20g/L).
Seed culture: the 50 mL MRS nutrient solutions of packing in the 250 mL triangular flasks, the lactic acid bacteria of picking purifying is inserted the MRS nutrient solution, seals with preservative film, leaves standstill in 30 ℃ of anaerobism and cultivates 18 h.
(3) fermentation condition: seed liquor inoculum concentration (v/v) is 4%, and the orange juice fermentation medium leaves standstill 35 ℃ of following anaerobism cultivates 130h, initial pH=6.4～6.6.The naringinase that adds 1100 U/L fermentation mediums during anaerobic fermentation to 70 h carries out synchronous debitterize (the naringinase vigor is 20000U/g).
After (4) 35 ℃ of anaerobism leave standstill and cultivate 130 h, adopt the GABA content in the liquid chromatography for measuring orange juice culture medium, draw GABA output and reach 3.8 g/L.Adopt coating orange juice solid fermentation medium therapy to record that the Lactobacillus plantarum viable count reaches 2 * 10 in the orange juice zymotic fluid again
8Cfu/ml, the sodium glutamate utilization rate reaches 70.9%.
(3) post processing:
The concentrate of orange juice culture medium is carried out spray-drying processing, and input concentration is about 40%, 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.
Dry back citrus fruit powder gets yield and is: 10%, and moisture≤4%.Viable count reaches 1.5 * 10
9Cfu/g, GABA content are 3.1%.
Embodiment 3:
(A) raw material preparation: In November picking citrus as raw materials, peeled juice, take 50kg sauce added cellulase and pectinase enzyme mass ratio of 3:1 powder (cellulase: 10000u / g , acid pectinase: activity 30000u / g, were purchased from Wuxi Xuemei preparations Ltd.) 120g of the enzyme treatment to obtain a crude orange juice, and then centrifuged to remove the bladder, the nucleolus and other debris, and then wrap with plastic membrane seal placed at 4 ℃ refrigerator cold standby.The allotment dilution of orange juice: glucose content is not 20.5 g/L when not diluting in the orange juice, dilutes 6 times, and sugared content is 3.35g/L.
(2) allotment of orange juice medium component and fermentation procedure
(1) adds yeast extract 20 g/L again to above-mentioned dilution orange juice, sodium acetate 2 g/L, sodium glutamate 15 g/L.Medium pH is transferred to 6.4～6.6, and 100 ℃ of sterilization 20min are cooled to room temperature.
(2) preservation of bacterial classification and cultural method:
With embodiment 1.
(3) fermentation condition: seed liquor inoculum concentration (v/v) is 4%, and the orange juice fermentation medium leaves standstill 35 ℃ of following anaerobism cultivates 100h, initial pH=6.4～6.6.The naringinase that adds 500 U/L fermentation mediums during anaerobic fermentation to 80 h carries out synchronous debitterize (the naringinase vigor is 20000U/g).
(4) 35 ℃ leave standstill cultivate 100 h after, adopt the GABA content in the liquid chromatography for measuring orange juice culture medium, draw GABA output and reach 4.677 g/L.Adopt coating orange juice solid fermentation medium therapy to record that the Lactobacillus plantarum viable count reaches 3 * 10 in the orange juice zymotic fluid again
8Cfu/ml, the sodium glutamate utilization rate reaches 72.3%.
(3) post processing:
The concentrate of orange juice culture medium is carried out spray-drying processing, and input concentration is about 40%, 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.
Dry back citrus fruit powder gets yield and is: 9%, and moisture≤4%.Viable count reaches 2.5 * 10
9Cfu/g, GABA content are 4.5%
Embodiment 4:
(1) raw material is prepared:
In November picking citrus as raw materials, peeled juice, take 50kg sauce added cellulase and pectinase enzyme mass ratio of 3:1 powder (cellulase: 10000u / g, acid pectinase: activity 30000u / g, were purchased from Wuxi Xuemei preparations Ltd.) 120g of the enzyme treatment to obtain a crude orange juice, and then centrifuged to remove the bladder, the nucleolus and other debris, and then sealed with plastic wrap and put into four ℃ in the refrigerator cold standby.The allotment dilution of orange juice: glucose content is not 35.5 g/L when not diluting in the orange juice, dilutes 6 times, and sugared content is 5.35g/L.
(2) allotment of orange juice medium component and fermentation procedure
(1) adds yeast extract 20 g/L again to above-mentioned dilution orange juice, sodium acetate 2 g/L, sodium glutamate 8 g/L.Medium pH is transferred to 6.4～6.6, and 100 ℃ of sterilization 20min are cooled to room temperature.
(2) preservation of bacterial classification and cultural method:
With embodiment 1.
(3) fermentation condition: seed liquor inoculum concentration (v/v) is 4%, and the orange juice fermentation medium leaves standstill 35 ℃ of following anaerobism cultivates 120h, initial pH=6.4～6.6.The naringinase that adds 650 U/L fermentation mediums during anaerobic fermentation to 80 h carries out synchronous debitterize (the naringinase vigor is 20000U/g).
(4) 35 ℃ leave standstill cultivate 120 h after, adopt the GABA content in the liquid chromatography for measuring orange juice culture medium, draw GABA output and reach 5.337 g/L.Adopt coating orange juice solid fermentation medium therapy to record that the Lactobacillus plantarum viable count reaches 3.9 * 10 in the orange juice zymotic fluid again
8Cfu/ml, the sodium glutamate utilization rate reaches 60.9%.
(3) post processing:
The concentrate of orange juice culture medium is carried out spray-drying processing, and input concentration is about 40%, 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.
Dry back citrus fruit powder gets yield and is: 11%, and moisture≤4%.Viable count reaches 3.1 * 10
9Cfu/g, GABA content are 5.0%.
Embodiment 5:
(1) raw material is prepared:
The tomato of plucking with October is a raw material, removes the peel and squeezes the juice, and gets the complex enzyme powder (cellulase: 10000u/g that the 50kg juice adds cellulase and pectase mass ratio 3:1; Acid pectase: enzyme 30000u/g alive; Available from Wuxi snow plum preparation Science and Technology Ltd.) altogether 120g carry out enzyme and handle, obtain thick tomato juice, carry out centrifugal filtration then; To remove residues such as leather bag, kernel, seal the refrigerator of putting into 4 ℃ with preservative film then and refrigerate subsequent use.The allotment dilution of tomato juice: sugar content is not 20.5 g/L when not diluting in the tomato juice, dilutes 4 times, and sugared content is 5.15g/L.
(2) allotment of tomato juice medium component and fermentation procedure
(1) adds yeast extract 20 g/L again to above-mentioned dilution tomato juice, sodium acetate 2 g/L, sodium glutamate 15 g/L.Medium pH is transferred to 6.4～6.6, and 100 ℃ of sterilization time 20min are cooled to room temperature.
(2) preservation of bacterial classification and cultural method:
With embodiment 1.
(3) fermentation condition: seed liquor inoculum concentration (v/v) is 4%, and the tomato juice fermentation medium leaves standstill 35 ℃ of following anaerobism cultivates 120h, initial pH=6.4～6.6.
(4) 35 ℃ leave standstill cultivate 120 h after, adopt the GABA content in the liquid chromatography for measuring tomato juice culture medium, draw GABA output and reach 5.224 g/L.Adopt the coating counting method to record that the Lactobacillus plantarum viable count reaches 4.5 * 10 in the tomato juice zymotic fluid again
8Cfu/ml, the sodium glutamate utilization rate reaches 80.9%.
(3) post processing:
The concentrate of tomato juice culture medium is carried out spray-drying processing, and input concentration is about 40%, 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.
Dry back tomato meal gets yield and is: 10%, and moisture≤4%.Viable count reaches 4 * 10
9Cfu/g, GABA content are 5%.
Claims (6)
1. preparation method who is rich in the fermented type fruits and vegetables powder of GABA and Lactobacillus plantarum, said method comprises:
(1) with raw material fruits and vegetables peeling squeezing, add cellulase and pectase complex enzyme powder and carry out the enzyme processing, centrifugal filtration is got supernatant and is obtained Juice again;
(2) Juice being diluted to sugar content is 2～50g/L, adds yeast extract 20 g/L again, sodium acetate 2 g/L, and glutamic acid or sodium glutamate 2～50 g/L, regulating the pH value is 6.6～6.8, obtains fermentation medium;
(3) fermentation medium is after sterilization, and inoculation Lactobacillus plantarum (Lactobacillus plantarum) CCTCC NO:M 2011364 leaves standstill cultivation 80～200h 30～35 ℃ of following anaerobism, obtains zymotic fluid;
(4) concentrated, the spray-drying of zymotic fluid promptly gets said fermented type fruits and vegetables powder.
2. the method for claim 1 is characterized in that raw material fruits and vegetables described in the step (1) are one of following: citrus, apple, grape, strawberry, tomato, watermelon.
3. the method for claim 1 is characterized in that middle cellulase of said step (1) and pectase enzyme powder consumption are 1:1 with enzyme work, and the complex enzyme total addition level is 2 ~ 50,000 U/kg Juices.
4. the method for claim 1; It is characterized in that in the said step (3); When raw materials used when being citrus; The naringinase that when anaerobic fermentation 60～150 h, adds 500 ~ 1200U/L fermentation medium carries out synchronous debitterize, and said naringinase is obtained through fermentation by aspergillus niger (Aspergillus niger) CCTCC NO:M 206047.
5. the method for claim 1 is characterized in that said step (4) spray-drying condition is: 35 ℃ of feeding temperatures, and 140 ℃ of inlet temperatures, 55 ℃ of outlet temperatures, air intake air pressure 0.25～0.3MPa, the centrifugal turntable rotating speed remains on 20000 r/min.
6. Lactobacillus plantarum (Lactobacillus plantarum) WZ011 is preserved in Chinese typical culture collection center, address: China; Wuhan, Wuhan University, postcode 430072; Preservation date on October 24th, 2011, deposit number CCTCC No:M 2011364.
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