(3) summary of the invention
The object of the invention provides a kind of with overproduction or the inferior fruits and vegetables of low value, as citrus, strawberry, tomato, apple, watermelon, grape, through the production technology of the eutrophy fruits and vegetables powder that complex enzyme is handled, microbial fermentation forms.By adopting new and high technologies such as enzyme engineering technology, fermentation and metabolic engineering technology, modern low temperature drying technology, effectively improve the content of vitamin, pectin oligosaccharide, amino acid etc. in the fruits and vegetables powder, improve the quality and the recovery rate of product, reduce production costs, can develop suitable marketing, having the medicinal of certain physiological function and edible natural vitamin fruit and vegetable food, is the desirable feedstock as food, medicine and health products.
The technical solution used in the present invention is:
A kind of processing method that is rich in the fermented type nutrient fruit and vegetable powder of vitamin, described method comprises:
(1) pre-treatment: get fruit and vegetable materials, clean, remove the peel, pull an oar, obtain the fruits and vegetables slurries; Described fruits and vegetables are one of following: citrus, apple, grape, strawberry, tomato, watermelon; Concrete grammar can be as follows: fruits and vegetables peeling back adds the water of 10~20 times of quality and processes preceding cleaning, removes the dirt of fruit and vegetable surfaces, with beater garden stuff processing is become thick fruits and vegetables slurries then.
(2) the phase I enzyme is handled: add cellulase and pectase complex enzyme powder in the fruits and vegetables slurries, adjust pH to 3.5~5.0 (citric acid or NaOH), at 40~45 ℃ of following stir process 1~4h; Described cellulase consumption is 30~50U/g pulp liquid, and the pectase consumption is 15~40U/g pulp liquid; Usually complex enzyme opaque amount consumption is 0.06%~0.16% of a fruits and vegetables slurries quality, wherein cellulase (30000~40000U/g) and pectase (ratio of 30000~50000U/g) quality is 2~3: 1; Need in the processing procedure constantly to stir, complete to accelerate enzymatic reaction;
(3) allotment: the fruits and vegetables slurries filter cleaner after step (2) is handled, get Juice, adding quality in the Juice is the nitrogenous source of Juice quality 1.5%~2.5%, and interpolation sucrose makes the soluble solid content of Juice reach 11Brix~19Brix, transfer pH to 4.5~5.0,111~121 ℃ sterilization 15~20min with citric acid or NaOH; Described nitrogenous source is a yeast extract;
(4) fermentation: in deployed Juice, insert saccharomycete, at 28~32 ℃, 120~350r/min bottom fermentation, 40~60h; Described saccharomycete is one of following or wherein two or more mixing: but salt tolerant flavouring yeast, Lang Bi Candida, KShi yeast, Shanghai are made yeast, grape wine high activity dried yeast and the super highly active dry yeast of 2.089 fruit wine; Zymotic fluid is concentrated into 15%~40% of original volume, gets concentrate; Inoculum concentration inoculum concentration routinely gets final product, and those of ordinary skills can determine that according to general knowledge with when enlarging the seed liquor inoculation after cultivating, seed liquor is 5~10% of a Juice volume usually;
The preservation of barms and expansion cultural method:
Slant preservation culture medium (100mL): peptone 1g, yeast extract 1g, NaCl 0.4g, glucose 1g, agar 2g, pH 6.5.30 ℃ of constant temperature culture 2~3 days, the preservation temperature is 4 ℃.
First order seed culture medium (100mL): glucose 10g, peptone 1g, yeast extract 1g, MgSO
47H
2O 0.02g, KH
2PO
40.02g pH 5.0.30 ℃ of cultivation temperature, shaking speed 200r/min, incubation time 18h.
Secondary seed jar culture medium (100mL): glucose 10g, peptone 1g, yeast extract 1g, MgSO
4H
2O 0.02g, KH
2PO
40.02g pH 5.0.30 ℃ of cultivation temperature, mixing speed 200r/min, incubation time 24h.
(5) post processing: the concentrate spray-drying obtains described fermented type nutrient fruit and vegetable powder.
When the raw material for preparing described fermented type nutrient fruit and vegetable powder is citrus, described method also includes second stage enzyme treatment step between step (4) and step (5): pH transfers to 4.1 with concentrate, add quality and be concentrate 0.3%~0.4% naringinase (10000~20000U/g), handle 1.5~2.5h down in 40 ℃ of constant temperature.
Preferably, described naringinase is obtained through fermentation by aspergillus niger CCTCC NO:M 206047 (aspergillus niger WZ001 (Aspergillus niger WZ001)), and this bacterial strain is being protected as new bacterial strain in first to file CN200610051963.X.
The debitterize enzyme fermentation both can adopt solid-state shallow tray fermentation also can adopt the deep liquid aerobic fermentation.
Solid-state shallow tray fermentation: wheat bran 100g; K
2HPO
40.3g; Bean dregs (weight in wet base, dry weight is about 12%) 60g; Soya-bean cake powder 8g; Orange meal 3.5g, water 200mL, pH nature.Condition of culture: inoculum concentration is 10%~15%, and 32 ℃ of cultivation temperature are turned over song behind the cultivation 30h, incubation time 3-4 days altogether.Oven dry, the dry place's preservation in sealing back, standby, naringinase enzyme activity 10000~20000U/g.。
Liquid ventilating fermentation (liquid amount by the 100L fermentation tank is 70L): bean dregs 7kg (weight in wet base, dry weight is about 12%); Soya-bean cake powder 1.75kg; Corn flour 0.7kg; Orange meal 0.7kg; K
2HPO
40.21kg; CaCl
270g; MgSO
47H
2O 70g; Defoamer (edible soybean oil) 50mL, pH 6.5.Condition of culture: cultivation temperature 30-32 ℃, incubation time 90~100h.Speed of agitator: preceding 20h is 150r/min, and DO (dissolved oxygen) is being got final product more than 30%; Control in real time through row progressively amplifying stirring frequency according to the dissolved oxygen reading behind the 20h.DO value minimum can not be lower than 10%, and is dry behind the filtering fermentation liquor, obtains the debitterize enzyme, naringinase enzyme activity 10000~20000U/g.
Described spray-drying condition is as follows: 55~60 ℃ of input concentration 30%~40%, temperature, 180~200 ℃ of inlet temperatures, 90~100 ℃ of outlet temperatures, air intake air pressure 0.25~0.3MPa, centrifugal turntable rotating speed 15000~20000r/min.
Beneficial effect of the present invention is mainly reflected in: according to the inventive method, make fruits and vegetables powder and traditional fruits and vegetables powder relatively, it is a kind of fully natural green product, nutritional labelings such as its vitamin, pectin oligosaccharide, amino acid significantly increase after the processing of operations such as enzyme processing, fermentation, wherein, content beta-carotene is 0.001%~0.05%, B family vitamin (VB
1, VB
2, nicotinic acid, VB
6Deng) in nicotinic acid, content is 0.2%~1.5%, pectin oligosaccharide content is 1%~10%; Dispersed and water-soluble all better in addition, moisture≤8% is more suitable for long-term preservation; It is composite to be with this fruits and vegetables powder that base stock carries out, and can produce composite nutritional products, is widely used in fields such as beverage, instant food, health products; Can increase the overall economic efficiency of fruits and vegetables industry on the whole, reduce the wasting of resources, protection environment, be the sustainable development generation deep effect of China's fruits and vegetables industry.
(4) specific embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1:
The preservation of barms is cultivated with expansion:
The slant preservation culture medium: peptone 1g, yeast extract 1g, NaCl 0.4g, glucose 1g, agar 2g, water 100mL, pH 6.5, high-temperature sterilization, cooling bevel.Insert bacterial classification, 30 ℃ of constant temperature culture 2~3 days, the preservation temperature is 4 ℃.
First order seed culture medium: glucose 10g, peptone 1g, yeast extract 1g, MgSO
47H
2O0.02g, KH
2PO
40.02g, water 100mL, pH 5.0.Inoculation one ring slant strains, 30 ℃ of cultivation temperature, shaking speed 200r/min, incubation time 18h gets primary seed solution.
Secondary seed jar culture medium: glucose 10g, peptone 1g, yeast extract 1g, MgSO
4H
2O 0.02g, KH
2PO
40.02g, water 100mL, pH 5.0.Insert primary seed solution, 30 ℃ of cultivation temperature, mixing speed 200r/min, incubation time 24h by 5% volume ratio.
Salt tolerant flavouring yeast (Jining Yu Yuan bio tech ltd) but, Lang Bi Candida (DSMZ of Microbe Inst., Chinese Academy of Sciences, preservation center numbering: As2.1182), KShi yeast (the Shanghai enlightening is sent out and brewageed biological products Co., Ltd), Shanghai yeast (the Shanghai enlightening is sent out and brewageed biological products Co., Ltd), grape wine high activity dried yeast (Angel Yeast Co.,Ltd) and the super highly active dry yeast (Angel Yeast Co.,Ltd) of making 2.089 fruit wine enlarge cultivation respectively according to the method described above, it is standby to obtain seed liquor respectively.
The fermentation of debitterize enzyme:
After 5 days, inoculation one ring is cultivated 36h to black-koji mould WZ001 (CCTCC NO:M 206047) bacterial classification to liquid seed culture medium (wheat bran 4g, soya-bean cake powder 3g, bean dregs 8g, water 100mL), obtain seed liquor through the activation of Cha Shi culture medium.
Solid-state fermentation culture medium is wheat bran 100g, bean dregs 10g, soya-bean cake powder 20g, the water that adds 100ml, be sub-packed in the triangular flask of 500ml, charging thickness is 2.5~3.0cm, 0.1Mpa steam pressure sterilization 30min, the aspergillus niger seed liquor is inserted with volume ratio 2% in every bottle of cooling back, cultivates 5~7 days down in 32 ℃, culture oven dry back is filtered to such an extent that debitterize enzyme liquid drying is standby by 1: 10 flooding, detects naringinase enzyme activity 10000~20000U/g in the enzyme powder.
Embodiment 2:
1. raw material is prepared: the Pon mandarin orange with the harvesting in November of area, Quzhou is a raw material, takes by weighing raw material 150kg after the selected classification, removes the peel the directly full pulp in back and pulls an oar, and gets thick citrus pulp 118.5kg, and it is standby to take out 50kg.
2. the thick citrus pulp of phase I enzyme treatment process: 50kg constantly stirs, and with NaOH pH is transferred to 4.2, and temperature rises to 42 ℃.Select that selectivity is strong for use, stability and good food-grade acid pectase and the cellulase of hydrolysis result be hybridly prepared into complex enzyme formulation, the mass ratio of cellulase and pectase is 3: 1.Add complex enzyme formulation 60g altogether in thick citrus pulp, enzyme processing time is 90min.Acid pectase: enzyme 30000u/g alive, available from Wuxi snow plum preparation Science and Technology Ltd.; Cellulase: enzyme 40000u/g alive, available from the grand mcroorganism Engineering Co., Ltd in Shandong.
With the thick citrus pulp behind the complex enzyme zymohydrolysis through plate and frame filter press press filtration (filter cloth 120 orders), remove leather bag in the thick citrus pulp and broken residues such as seed, orange juice 40.4kg, crushing juice rate is 80.8%.
Contrast test: other gets, and the thick citrus pulp of 50kg is not enzyme-added to compare, and directly with thick citrus pulp squeezing, gets 34.6kg, and crushing juice rate is 69.2%.
This shows that adding complex enzyme processing back crushing juice rate has increased by 11.7%.Can effectively improve the crushing juice rate of fruit processing by the catalytic reaction of pectase and cellulase, improve the fruit juice quality and yield, simple for process.Pectase can soften the pectic substance in the pulp organization, makes insoluble pectin substance be decomposed into the pectin oligomer of solubility, to reduce the viscosity of pulp, can obviously improve the fruit juice productive rate.Cellulase synergistic destroys the fibre structure in the fruits and vegetables, is the utilizable reductive monosaccharide of microorganism with cellulose degradation.The transparency of fruit juice and clarity are able to obvious raising, have also increased the content of soluble solid.
4. the allotment of orange juice and fermentation:
(1) allotment of orange juice: press mass fraction 1.8% and in the 40.4kg orange juice, add nitrogenous source yeast extract 0.73kg; The pol that detects orange juice with hand-held saccharometer is 13.5Brix, adds 2.70kg sucrose and make the orange juice pol reach 18Brix in the 40.4kg orange juice; With NaOH orange juice pH is transferred to 5.0.121 ℃ of sterilization 15min are cooled to room temperature.
(2) in the autocontrol mechanical agitating fermentation tank of 65L, the salt tolerant flavouring yeast starter liquid for preparing is moved into the deployed orange juice of 41L by the inoculum concentration of 10% (v/v), making liquid amount is 45L.Control tank pressure 0.03Mpa, bubbling air pressure 0.12Mpa, air velocity 25NL/min, 30 ℃ of temperature, mixing speed 300r/min, fermentation time are 50h.
5. be raw material with the citrus fruit, contain a large amount of bitter substances in the zymotic fluid, the main source of bitter taste is an aurantiin, therefore, removes aurantiin the fruit juice bitter taste is alleviated.The used enzyme of enzyme process debitterize mainly is a naringinase, and it is made up of alpha-L-Rhamnosidase and β-D-glucuroide.Alpha-L-Rhamnosidase can be hydrolyzed into prunin and rhamnose with the aurantiin of flavanones glucoside compound in the citrus fruit juice, and the bitter taste of prunin is about 1/3 of aurantiin, so bitter taste alleviates to some extent; Prunin can generate the naringenin and the glucose of no bitter taste under β-D-glucuroide continuation effect.It is that naringinase is handled that zymotic fluid is carried out the second step debitterize enzyme, and concrete grammar is:
Ratio in 20% is concentrated to 9kg with zymotic fluid, detects the about 2.15g/L of naringin content in the concentrate.The enzyme treatment conditions are:
(1) 40 ℃ of constant temperature of concentrate, pH transfers to 4.1.
(2) the enzyme addition needs to add embodiment 1 debitterize enzyme (naringinase enzyme activity 10000U/g) enzyme powder 27g for concentrating 0.3% of after fermentation liquid quality, and the debitterize time is 2h.
6. the concentrate after the debitterize enzyme is handled is carried out spray-drying processing, input concentration is about 40%, 60 ℃ of feeding temperatures, and 180~200 ℃ of inlet temperatures, 90~100 ℃ of outlet temperatures, air intake air pressure 0.25~0.3MPa, the centrifugal turntable rotating speed remains on 18000r/min.
7. obtain fermented type eutrophy citrus grain weight 3.61kg after the drying, recovery rate is: 8.94% (accounting for orange juice proportion after filtering), moisture≤4%.Various vitamin contents such as following table 1 in the fermented type eutrophy citrus powder.Sealing after cooling places 4 ℃ of dry place's preservations.
Table 1: several vitamins content (100g) in the fermented type eutrophy citrus powder
Embodiment 3:
1. raw material is prepared: with the regional Pon mandarin orange in the Quzhou of embodiment 2 is raw material, chooses 150kg, and the full pulp making beating in peeling back gets thick citrus pulp 116.0kg, and it is standby to take out 50kg.
2. phase I enzyme treatment process: the thick citrus pulp of 50kg according to embodiment 2 step regulatory enzyme reaction conditions, is added complex enzyme formulation, and the ratio of cellulase and pectase is 2: 1.Add complex enzyme formulation 70g in thick citrus pulp, enzyme processing time is 90min.Acid pectase: enzyme 30000u/g alive, available from Wuxi snow plum preparation Science and Technology Ltd.; Cellulase: enzyme 40000u/g alive, available from the grand mcroorganism Engineering Co., Ltd in Shandong.
3. the filtration operation according to embodiment 2 gets orange juice 42.1kg, and crushing juice rate is 84.2%.Contrast test: other gets, and the thick citrus pulp of 50kg is not enzyme-added to compare, and directly with thick citrus pulp squeezing, gets 34.6kg, and crushing juice rate is 69.2%.This shows that adding complex enzyme processing back crushing juice rate has increased by 15.0%.
4. the allotment of orange juice and fermentation:
(1) allotment of orange juice: press mass fraction 2.0% and add nitrogenous source yeast extract 0.84kg in the 42.1kg orange juice, the pol that detects orange juice with hand-held saccharometer is 13.8Brix, adds 2.67kg sucrose and make the orange juice pol reach 18Brix in the 42.1kg orange juice; With NaOH orange juice pH is transferred to 5.0.121 ℃ of sterilization 15min are cooled to room temperature.
(2) in the autocontrol mechanical agitating fermentation tank of 65L, but brightly must the Candida seed liquor move into the deployed orange juice of 42L by the inoculum concentration of 10% (v/v) with what prepare, making liquid amount is 46L.Control tank pressure 0.03Mpa, bubbling air pressure 0.12Mpa, air velocity 25NL/min, 30 ℃ of temperature, mixing speed 300r/min, fermentation time are 50h.
5. zymotic fluid is carried out the second step debitterize enzyme and handle, the ratio in 20% is concentrated to 9.2kg with zymotic fluid, detects the about 2.22g/L of naringin content in the concentrate.The enzyme treatment conditions: 40 ℃ of constant temperature, pH are 4.1.Debitterize enzyme (naringinase enzyme activity 10000U/g) enzyme powder addition adds 27g approximately for concentrating 0.3% of after fermentation liquid quality, and the debitterize time is 2h.
6. carry out spray drying treatment according to 2 pairs of concentrates of embodiment, obtain dry after fermentation type eutrophy citrus grain weight 3.80kg, recovery rate is: 9.05% (accounting for orange juice proportion after filtering), moisture≤4%.Various vitamin contents such as following table 2 in the fermented type eutrophy citrus powder.Sealing after cooling places 4 ℃ of dry place's preservations.
Table 2: several vitamins content (100g) in the fermented type eutrophy citrus powder
Embodiment 4:
1. raw material is prepared: with the regional Pon mandarin orange in the Quzhou of embodiment 2 is raw material, chooses 150kg, and the full pulp making beating in peeling back gets thick citrus pulp 108.8kg, and it is standby to take out 50kg.
2. phase I enzyme treatment process: the thick citrus pulp of 50kg according to embodiment 2 step regulatory enzyme reaction conditions, is added complex enzyme formulation, and the ratio of cellulase and pectase is 2: 1.Add complex enzyme formulation 70g in thick citrus pulp, enzyme processing time is 90min.Acid pectase: enzyme 30000u/g alive, available from Wuxi snow plum preparation Science and Technology Ltd.; Cellulase: enzyme 40000u/g alive, available from the grand mcroorganism Engineering Co., Ltd in Shandong.
3. the filtration operation according to embodiment 2 gets orange juice 41.4kg, and crushing juice rate is 82.8%.Contrast test: other gets, and the thick citrus pulp of 50kg is not enzyme-added to compare, and directly with thick citrus pulp squeezing, gets 34.6kg, and crushing juice rate is 69.2%.This shows that adding complex enzyme processing back crushing juice rate has increased by 13.6%.
4. the allotment of orange juice and fermentation:
(1) allotment of orange juice: press mass fraction 1.7% and in the 41.4kg orange juice, add nitrogenous source yeast extract 0.70kg; The pol that detects orange juice with hand-held saccharometer is 13.5Brix, adds 2.82kg sucrose and make the orange juice pol reach 19Brix in the 41.4kg orange juice; With NaOH orange juice pH is transferred to 5.0.121 ℃ of sterilization 15min are cooled to room temperature.
(2) in the autocontrol mechanical agitating fermentation tank of 65L, the KShi yeast starter liquid for preparing is moved into the deployed orange juice of 41L by the inoculum concentration of 7% (v/v), making liquid amount is 45L.Control tank pressure 0.03Mpa, bubbling air pressure 0.12Mpa, air velocity 25NL/min, 30 ℃ of temperature, mixing speed 300r/min, fermentation time are 52h.
5. zymotic fluid is carried out the second step debitterize enzyme and handle, the ratio in 20% is concentrated to 9.0kg with zymotic fluid, detects the about 2.14g/L of naringin content in the concentrate.The enzyme treatment conditions: 40 ℃ of constant temperature, pH are 4.1.Debitterize enzyme (naringinase enzyme activity 10000U/g) enzyme powder addition adds 27g approximately for concentrating 0.3% of after fermentation liquid quality, and the debitterize time is 2h.
6. carry out spray drying treatment according to 2 pairs of concentrates of embodiment, obtain dry after fermentation type eutrophy citrus grain weight 3.72kg, recovery rate is: 8.98% (accounting for orange juice proportion after filtering), moisture≤4%.Various vitamin contents such as following table 3 in the fermented type eutrophy citrus powder.Sealing after cooling places 4 ℃ of dry place's preservations.
Table 3: several vitamins content (100g) in the fermented type eutrophy citrus powder
Embodiment 5:
1. raw material is prepared: taking by weighing clean, sorting after ripening degree is the fresh tomato 110kg 9.0 or more, full fruit making beating, and the thick strained tomatoes of taking-up 50kg is standby.
2. the thick strained tomatoes of phase I enzyme treatment process: 50kg constantly stirs, and with citric acid pH is transferred to 4.2, and temperature rises to 42 ℃.Select that selectivity is strong for use, stability and good food-grade acid pectase and the cellulase of hydrolysis result be hybridly prepared into complex enzyme formulation, the ratio of cellulase and pectase is 3: 1.Add complex enzyme formulation 40g in thick strained tomatoes, enzyme processing time is 60min.Acid pectase: enzyme activity 30000u/g, available from Wuxi snow plum preparation Science and Technology Ltd.; Cellulase: enzyme activity 40000u/g, available from the grand mcroorganism Engineering Co., Ltd in Shandong.
With the thick strained tomatoes behind the complex enzyme zymohydrolysis through plate and frame filter press press filtration (filter cloth 120 orders), remove leather bag in the thick strained tomatoes and broken residues such as seed, tomato juice 43.7kg, crushing juice rate is 87.4%.
Contrast test: other gets, and the thick strained tomatoes of 50kg is not enzyme-added to compare, and directly with thick strained tomatoes squeezing, gets 40.9kg, and crushing juice rate is 81.8%.This shows that adding complex enzyme processing back crushing juice rate has increased by 5.6%.
4. the allotment of tomato juice and fermentation:
(1) allotment of tomato juice: press mass fraction 1.8% and add nitrogenous source yeast extract 0.78kg in the 43.7kg orange juice, the pol that detects tomato juice with hand-held saccharometer is 4.5Brix, adds 3.8kg sucrose and make pol reach 12.5Brix in tomato juice; With NaOH orange juice pH is transferred to 5.0.121 ℃ of sterilization 15min are cooled to room temperature.
(2) in the autocontrol mechanical agitating fermentation tank of 65L, the super brewer yeast seed liquor for preparing is moved into the deployed tomato juice of 44L by the inoculum concentration of 10% (v/v), making liquid amount is 48L.Control tank pressure 0.03Mpa, bubbling air pressure 0.13Mpa, air velocity 27NL/min, 30 ℃ of temperature, mixing speed 330r/min, fermentation time are 42h.
5. in 30% ratio zymotic fluid is concentrated to 14.4kg, concentrate is carried out spray-drying processing, input concentration about 40%, 60 ℃ of feeding temperatures, 180~200 ℃ of inlet temperatures, 90~100 ℃ of outlet temperatures, air intake air pressure 0.25~0.3MPa, the centrifugal turntable rotating speed remains on 20000r/min.
6. obtain fermented type eutrophy tomato grain weight 5.89kg after the drying, recovery rate is: 13.47% (accounting for tomato juice proportion after filtering), moisture≤5%.Various vitamin contents such as following table 4 in the fermented type eutrophy tomato meal.Sealing after cooling places 4 ℃ of dry place's preservations.
Table 4: several vitamins content (100g) in the fermented type eutrophy tomato meal