CN106318999A - Method for preparing pumpkin seed meal molecular peptide through solid state fermentation on pumpkin seed cake meal by virtue of mixed bacteria - Google Patents

Method for preparing pumpkin seed meal molecular peptide through solid state fermentation on pumpkin seed cake meal by virtue of mixed bacteria Download PDF

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CN106318999A
CN106318999A CN201610706267.1A CN201610706267A CN106318999A CN 106318999 A CN106318999 A CN 106318999A CN 201610706267 A CN201610706267 A CN 201610706267A CN 106318999 A CN106318999 A CN 106318999A
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fermentation
semen cucurbitae
powder
aspergillus niger
pumpkin seed
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丁舸
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YIWU JIANGR BIO-TECHNOLOGY Co Ltd
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YIWU JIANGR BIO-TECHNOLOGY Co Ltd
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products

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Abstract

The invention discloses a method for preparing pumpkin seed meal molecular peptide through solid state fermentation on pumpkin seed cake meal by virtue of mixed bacteria. The method comprises the following steps: (1) fermenting lactic acid bacteria; (2) fermenting Aspergillus niger; and (3) extracting and separating the pumpkin seed meal molecular peptide: mixing the fermenting products with water in a weight ratio of 1 to (20-25), stirring and soaking at 55-60 DEG C for 4-6 hours, carrying out plate-frame pressure filtration to intercept filtrate by virtue of a hollow fiber column with a molecular mass of 10000 Daltons so as to obtain ultrafiltrate, carrying out reduced pressure concentration on the ultrafiltrate, adsorbing the ultrafiltrate by virtue of a macroporous resin column, carrying out elution, and carrying out educed pressure concentration and spray drying on eluate, so as to obtain the pumpkin seed meal molecular peptide. The process of the method is simple and feasible and is applicable to industrial production, the cost is low, the conversion rate of the pumpkin seed meal molecular peptide is high, the taste and flavor of the pumpkin seed meal molecular peptide are greatly improved, and the obtained pumpkin seed meal molecular peptide is easy to absorb.

Description

A kind of method that Semen Cucurbitae dregs of rice molecular peptide prepared by mixed vaccine solid fermentation Semen Cucurbitae grouts
Technical field
A kind of method that the present invention relates to Semen Cucurbitae dregs of rice molecular peptide, particularly to a kind of mixed vaccine solid fermentation Fructus Cucurbitae moschatae seedcake The dregs of rice prepare the method for Semen Cucurbitae dregs of rice molecular peptide.
Background technology
Containing thick protein 70%-75%, crude fat 5%-7%, crude fibre 5%-6% in Semen Cucurbitae grouts.In addition, Fructus Cucurbitae moschatae Seedcake dregs of rice Mineral Elements and amino acid content are the most more.After Semen Cucurbitae oil expression, in addition to fat content reduces, other nutritional labelings Substantially all it is retained in Semen Cucurbitae grouts, but Semen Cucurbitae grouts are typically cooked feedstuff and process by country facilities, do not play it Potential economic worth.
Peptide is the form of a kind of special molecular weight of various protein, but it is special significantly to change its function by hydrolysis Property.Along with its degree of hydrolysis increases, water solublity is consequently increased, and emulsibility and foaming characteristic all first increase with the increase of degree of hydrolysis again Reducing, its viscosity declines, and external digestion sex index increases with degree of hydrolysis and improves, and the peptide of this kind of easy absorption can be sought as intestinal Foster agent becomes the people that spoon meat form is supplied under special condition.Semen Cucurbitae grouts molecular peptide has regulation blood pressure, reduces The functions such as cholesterol, promotes lipid metabolism, improves immunity of organisms, slow down aging, and kinds of tumor cells is had induction Differentiation, apoptosis-induced and Inhibit proliferaton effect, widely, market potential is huge for its application prospect.
At present, the production of Semen Cucurbitae dregs of rice molecular peptide is scarcely out of swaddling-clothes by China, is concentrated mainly on enzymatic hydrolysis Fructus Cucurbitae moschatae Seed cake protein powder, this type of processing method not only cost is high, and (some produced after enzymolysis are special also to produce a certain degree of bitterness Oligopeptide bitter peptides), hardly result in popularization at food processing field.
Summary of the invention
It is an object of the invention to provide a kind of mixed vaccine solid fermentation Semen Cucurbitae grouts and prepare Semen Cucurbitae dregs of rice molecular peptide Method, simple for process, it is suitable for industrialized production, low cost, the conversion ratio of Semen Cucurbitae dregs of rice molecular peptide is high, greatly improves south The taste and flavor of melon seed dregs of rice molecular peptide, gained Semen Cucurbitae dregs of rice molecular peptide is prone to absorb.
The technical solution adopted for the present invention to solve the technical problems is:
A kind of method that Semen Cucurbitae dregs of rice molecular peptide prepared by mixed vaccine solid fermentation Semen Cucurbitae grouts, comprises the steps:
(1) lactic acid bacteria fermentation: lactic acid bacteria and sterilized rice flour according to the weight ratio of 1:4-6 mix lactic acid bacteria rice flour mixes Thing, puts into mix homogeneously in fermentation medium by lactic acid bacteria rice flour mixture, and lactic acid bacteria rice flour amount of mixture is fermentation culture The 10-15% of basic weight amount, 35-40 DEG C of fermentation culture；Put into fermentation medium with sterilized rice flour after being mixed by lactic acid bacteria again to send out Ferment, so inoculation are more uniform, increase carbon source simultaneously, improve Lactobacillus Survival high, and ferment effect is good；Send out through lactic acid bacteria Ferment, reduces the pH value of fermentation medium so that it is be more suitable for Aspergillus Niger Growth.The bacteriocin lab that lactic acid bacteria produces, has and presses down Make and kill most Gram-positive pathogenic bacteriums and putrefaction bacteria in fermentation medium.Suppression miscellaneous bacteria and pathogenic bacterium growth.Meanwhile, breast The decomposition of Semen Cucurbitae cake protein is played an important role by a series of enzymes that acid bacterium produces.
(2) fermentation of Aspergillus niger: when the pH of lactic acid bacteria fermentation cultivation to fermentation medium is down to 4.5-5.0, access black fermented preparation Mould spore powder, Aspergillus niger spores powder consumption is the 15-18% of fermentation medium weight, 32-35 DEG C of fermentation culture 70-72 hour Tunning, the blowing air amount that fermentation of Aspergillus niger is cultivated first 24 hours is 4000-4500m3/ h, fermentation culture 25-49 hour Blowing air amount is 2500-3500m3/ h, when fermentation culture to 50h, adds fermentation medium weight 15-in fermentation medium The nutritional solution of 20% weight, control blowing air amount is 900-1100m simultaneously3/ h is to fermentation ends；Logical by controlling specific fermentation The change of tolerance, ferment effect is good, and the conversion ratio of Semen Cucurbitae dregs of rice molecular peptide is high, greatly improves the mouthfeel of Semen Cucurbitae dregs of rice molecular peptide And local flavor；
(3) extraction of Semen Cucurbitae dregs of rice molecular peptide separates: the weight ratio that tunning is pressed 1:20-25 mixes with water, and 55-60 DEG C is stirred Mixing immersion 4-6 hour, through filter press, filtrate is filtered through the daltonian hollow fiber column of trapped molecular weight 10000 and is retained, Ultrafiltrate, ultrafiltrate is through concentrating under reduced pressure, and upper macroporous resin column is adsorbed, eluting, after eluent concentrating under reduced pressure, is spray-dried, obtains southern Melon seed dregs of rice molecular peptide.
The present invention opens the brand-new microorganism solid fermentation hydrolysis Semen Cucurbitae grouts that utilize of knowing clearly and prepares Semen Cucurbitae dregs of rice molecular peptide, The most with low cost, and the bitterness brought due to hydrophobic amino acid can be removed.
The present invention uses lactic acid bacteria, aspergillus niger mixed fermentation to prepare Semen Cucurbitae dregs of rice molecular peptide.First by lactobacillus inoculum in steaming In the well-done Semen Cucurbitae dregs of rice, the PH of Semen Cucurbitae dregs of rice substrate is down to the optimal condition of fermentation of Aspergillus niger；Again aspergillus niger is inoculated in Semen Cucurbitae dregs of rice substrate carries out mixed fermentation.In this sweat, along with the change of PH, aspergillus niger can produce basic protein Enzyme, neutral protease and acid protease, use it as liquid of protease and have more restriction enzyme site than general merchandise, can be significantly Improve percent hydrolysis.Meanwhile, by the effect of microorganism to some picryl because modifying and shift, recombinating, greatly improve The taste and flavor of molecular peptide.During the fermentation, Semen Cucurbitae cake protein through metabolism, assimilation, is converted again by little peptide ammino acid Peptide is moved in logistics of making a living, it is easy to absorb.
As preferably, described in step (1), fermentation medium forms as follows by weight percentage: Semen Cucurbitae grouts powder 70%, Semen Cucurbitae shell powder 10%, water 20%；PH is natural, 121 DEG C, sterilizing half an hour, is cooled to 35-40 DEG C.Use Semen Cucurbitae grouts Powder and Semen Cucurbitae shell powder are as fermentation medium, and such ferment effect is good, also can effectively utilize Semen Cucurbitae shell.
As preferably, described Semen Cucurbitae grouts powder is to obtain after Semen Cucurbitae grouts are pulverized, and Semen Cucurbitae grouts powder size is at 40- 60 mesh, the size of Semen Cucurbitae shell powder is at 20-40 mesh.
As preferably, in step (1), the manufacture method of lactic acid bacteria is: in 500L water add 50-70kg whole milk powder and 45-55kg sucrose, heated and stirred dissolves, 115 DEG C, sterilizing 30-60min under 0.1MPa, is cooled to 40-42 DEG C, is subsequently adding 5- 10kg lactobacillus powder stirs, and keeps little fermentation culture 8-12 hour of temperature 40-42 DEG C.
As preferably, in step (2), the manufacture method of Aspergillus niger spores powder is: by aspergillus niger slant strains sterilized water Washing out, making Aspergillus niger spores concentration is 2 × 106 -7×106The Aspergillus niger spores suspension of individual/mL, Aspergillus niger spores suspension Adding in activation medium, cultivate 24-26 hour for 35-38 DEG C, then the Semen Cucurbitae shell powder with sterilizing mixes according to 1:1.5- The part by weight of 2.5 mixes to obtain Aspergillus niger spores powder.
As preferably, the raw material weight percentage ratio of described activation medium consists of: wheat bran 56%, Semen Cucurbitae grouts powder 10%、NaH2PO42%、MgSO41%、NaNO30.5%, yeast extract 0.5%, water 30%, pH is natural, 121 DEG C, sterilizing half an hour, cooling To 35-40 DEG C.
As preferably, step (2) Middle nutrition liquid formula by weight percentage is: concentration of glucose 3%, biphosphate Potassium 1.6%, nicotinic acid 2.2 ‰, choline 1.8 ‰, folic acid 2.23 ‰, VB22.8 ‰, water surplus.
As preferably, in step (3), the resin of macroporous resin column is macroporous resin D3520, macroporous resin column absorption, eluting Technological parameter be: sample with macroporous resin column on the flow velocity of 2BV/h, then with 20% ethanol solution rinse to effluent be nothing Use 70% ethanol solution eluting during color instead, collect the eluent of 70% ethanol solution eluting.
As preferably, the flow velocity that 20% ethanol solution rinses is 4BV/h, and the elution speed of 70% ethanol solution eluting is 2BV/ h。
As preferably, the ventilation temperature being spray-dried in step (3) is 210 DEG C, and leaving air temp 95 DEG C, feed rate is 30L/h。
The invention has the beneficial effects as follows: simple for process, be suitable for industrialized production, low cost, Semen Cucurbitae dregs of rice molecular peptide Conversion ratio high, greatly improve the taste and flavor of Semen Cucurbitae dregs of rice molecular peptide, gained Semen Cucurbitae dregs of rice molecular peptide is prone to absorb.
Detailed description of the invention
Below by specific embodiment, technical scheme is described in further detail.
In the present invention, if not refering in particular to, the raw material used and equipment etc. are all commercially available or commonly used in the art. Method in following embodiment, if no special instructions, is the conventional method of this area.
Embodiment 1:
A kind of method that Semen Cucurbitae dregs of rice molecular peptide prepared by mixed vaccine solid fermentation Semen Cucurbitae grouts, comprises the steps:
(1) lactic acid bacteria fermentation: lactic acid bacteria and sterilized rice flour according to the weight ratio of 1:4 mix lactic acid bacteria rice flour mixes Thing, puts into mix homogeneously in fermentation medium by lactic acid bacteria rice flour mixture, and lactic acid bacteria rice flour amount of mixture is fermentation culture The 10% of basic weight amount, 35-40 DEG C of fermentation culture；Described fermentation medium forms as follows by weight percentage: Semen Cucurbitae grouts Powder (40-60 mesh) 70%, Semen Cucurbitae shell powder (20-40 mesh) 10%, water 20%；PH is natural, 121 DEG C, sterilizing half an hour, is cooled to 35- 40℃.The manufacture method of lactic acid bacteria is: add 50kg whole milk powder and 45kg sucrose in 500L water, and heated and stirred is dissolved, 115 DEG C, sterilizing 30-60min under 0.1MPa, be cooled to 40 DEG C, be subsequently adding 5kg lactobacillus powder (commercially available) and stir, keep temperature 40 DEG C little fermentation culture obtain Yoghourt shape product in 12 hours.
(2) fermentation of Aspergillus niger: when the pH of lactic acid bacteria fermentation cultivation to fermentation medium is down to 5.0, access aspergillus niger spore Sub-powder, Aspergillus niger spores powder consumption is the 15% of fermentation medium weight, and 32 DEG C of fermentation culture obtain tunning, black fermented preparation in 72 hours The mould fermentation culture blowing air amount of first 24 hours is 4000m3/ h, the blowing air amount of fermentation culture 25-49 hour is 2500m3/ h, When fermentation culture to 50h, in fermentation medium, add the nutritional solution of fermentation medium weight 15% weight, control logical simultaneously Air capacity is 900m3/ h is to fermentation ends.
The manufacture method of Aspergillus niger spores powder is: is washed out by aspergillus niger slant strains sterilized water, makes Aspergillus niger spores Concentration is 7 × 106The Aspergillus niger spores suspension of individual/mL, adds to Aspergillus niger spores suspension in activation medium, cultivates 26 for 35 DEG C Hour, then the Semen Cucurbitae shell powder with sterilizing mixes and mixes to obtain Aspergillus niger spores powder according to the part by weight of 1:1.5.Activation training The raw material weight percentage ratio supporting base consists of: wheat bran 56%, Semen Cucurbitae grouts powder 10%, NaH2PO42%、MgSO41%、 NaNO30.5%, yeast extract 0.5%, water 30%, pH is natural, 121 DEG C, sterilizing half an hour, be cooled to 35-40 DEG C.
Nutritional solution formula by weight percentage is: concentration of glucose 3%, potassium dihydrogen phosphate 1.6%, nicotinic acid 2.2 ‰, gallbladder Alkali 1.8 ‰, folic acid 2.23 ‰, VB22.8 ‰, water surplus.
(3) extraction of Semen Cucurbitae dregs of rice molecular peptide separates: the weight ratio that tunning is pressed 1:20 mixes with water, 55 DEG C of stirrings Soaking 6 hours, through filter press, filtrate is filtered through the daltonian hollow fiber column of trapped molecular weight 10000 and is retained, and obtains ultrafiltration Liquid, ultrafiltrate is through concentrating under reduced pressure, and sample adsorbs with macroporous resin column on the flow velocity of 2BV/h (macroporous resin D3520), then with 4BV/h flow velocity, uses 70% ethanol solution eluting instead when being colourless with 20% ethanol solution flushing to effluent, and 70% ethanol solution is washed De-elution speed is 2BV/h, collects the eluent of 70% ethanol solution eluting, after eluent concentrating under reduced pressure, is spray-dried, Semen Cucurbitae dregs of rice molecular peptide.The ventilation temperature being spray-dried is 210 DEG C, and leaving air temp 95 DEG C, feed rate is 30L/h.
Embodiment 2
A kind of method that Semen Cucurbitae dregs of rice molecular peptide prepared by mixed vaccine solid fermentation Semen Cucurbitae grouts, comprises the steps:
(1) lactic acid bacteria fermentation: lactic acid bacteria and sterilized rice flour according to the weight ratio of 1:6 mix lactic acid bacteria rice flour mixes Thing, puts into mix homogeneously in fermentation medium by lactic acid bacteria rice flour mixture, and lactic acid bacteria rice flour amount of mixture is fermentation culture The 15% of basic weight amount, 35-40 DEG C of fermentation culture；Described fermentation medium forms as follows by weight percentage: Semen Cucurbitae grouts powder (40-60 mesh) 70%, Semen Cucurbitae shell powder (20-40 mesh) 10%, water 20%；PH is natural, 121 DEG C, sterilizing half an hour, is cooled to 35-40 ℃.The manufacture method of lactic acid bacteria is: add 70kg whole milk powder and 55kg sucrose in 500L water, and heated and stirred is dissolved, 115 DEG C, sterilizing 30-60min under 0.1MPa, be cooled to 42 DEG C, be subsequently adding 10kg lactobacillus powder (commercially available) and stir, keep temperature Spend 42 DEG C little fermentation culture and obtain Yoghourt shape product in 8 hours.
(2) fermentation of Aspergillus niger: when the pH of lactic acid bacteria fermentation cultivation to fermentation medium is down to 4.5, access aspergillus niger spore Sub-powder, Aspergillus niger spores powder consumption is the 18% of fermentation medium weight, and 35 DEG C of fermentation culture obtain tunning, black fermented preparation in 70 hours The mould fermentation culture blowing air amount of first 24 hours is 4500m3/ h, the blowing air amount of fermentation culture 25-49 hour is 3500m3/ h, When fermentation culture to 50h, in fermentation medium, add the nutritional solution of fermentation medium weight 20% weight, control logical simultaneously Air capacity is 1100m3/ h is to fermentation ends.
The manufacture method of Aspergillus niger spores powder is: is washed out by aspergillus niger slant strains sterilized water, makes Aspergillus niger spores Concentration is 2 × 106The Aspergillus niger spores suspension of individual/mL, adds to Aspergillus niger spores suspension in activation medium, 38 DEG C of cultivations 24 hours, then the Semen Cucurbitae shell powder with sterilizing mixed and mixes to obtain Aspergillus niger spores powder according to the part by weight of 1:2.5.Live The raw material weight percentage ratio changing culture medium consists of: wheat bran 56%, Semen Cucurbitae grouts powder 10%, NaH2PO42%、MgSO41%、 NaNO30.5%, yeast extract 0.5%, water 30%, pH is natural, 121 DEG C, sterilizing half an hour, be cooled to 35-40 DEG C.
Nutritional solution formula by weight percentage is: concentration of glucose 3%, potassium dihydrogen phosphate 1.6%, nicotinic acid 2.2 ‰, gallbladder Alkali 1.8 ‰, folic acid 2.23 ‰, VB22.8 ‰, water surplus.
(3) extraction of Semen Cucurbitae dregs of rice molecular peptide separates: the weight ratio that tunning is pressed 1:25 mixes with water, and 60 DEG C are stirred Mixing immersion 4 hours, through filter press, filtrate is filtered through the daltonian hollow fiber column of trapped molecular weight 10000 and is retained, and obtains super Filtrate, ultrafiltrate is through concentrating under reduced pressure, and sample adsorbs with macroporous resin column on the flow velocity of 2BV/h (macroporous resin D3520), then with 4BV/h flow velocity, uses 70% ethanol solution eluting instead when being colourless with 20% ethanol solution flushing to effluent, and 70% ethanol solution is washed De-elution speed is 2BV/h, collects the eluent of 70% ethanol solution eluting, after eluent concentrating under reduced pressure, is spray-dried, Semen Cucurbitae dregs of rice molecular peptide.The ventilation temperature being spray-dried is 210 DEG C, and leaving air temp 95 DEG C, feed rate is 30L/h.
Embodiment 3:
A kind of method that Semen Cucurbitae dregs of rice molecular peptide prepared by mixed vaccine solid fermentation Semen Cucurbitae grouts, comprises the steps:
(1) lactic acid bacteria fermentation: lactic acid bacteria and sterilized rice flour according to the weight ratio of 1:5 mix lactic acid bacteria rice flour mixes Thing, puts into mix homogeneously in fermentation medium by lactic acid bacteria rice flour mixture, and lactic acid bacteria rice flour amount of mixture is fermentation culture The 12% of basic weight amount, 35-40 DEG C of fermentation culture；Described fermentation medium forms as follows by weight percentage: Semen Cucurbitae grouts Powder (40-60 mesh) 70%, Semen Cucurbitae shell powder (20-40 mesh) 10%, water 20%；PH is natural, 121 DEG C, sterilizing half an hour, is cooled to 35- 40℃.The manufacture method of lactic acid bacteria is: add 60kg whole milk powder and 50kg sucrose in 500L water, and heated and stirred is dissolved, 115 DEG C, sterilizing 30-60min under 0.1MPa, be cooled to 40 DEG C, be subsequently adding 8kg lactobacillus powder (commercially available) and stir, keep temperature 40 DEG C little fermentation culture obtain Yoghourt shape product in 10 hours.
(2) fermentation of Aspergillus niger: when the pH of lactic acid bacteria fermentation cultivation to fermentation medium is down to 4.8, access aspergillus niger spore Sub-powder, Aspergillus niger spores powder consumption is the 15% of fermentation medium weight, and 33 DEG C of fermentation culture obtain tunning, black fermented preparation in 70 hours The mould fermentation culture blowing air amount of first 24 hours is 4000m3/ h, the blowing air amount of fermentation culture 25-49 hour is 3000m3/ h, When fermentation culture to 50h, in fermentation medium, add the nutritional solution of fermentation medium weight 18% weight, control logical simultaneously Air capacity is 1000m3/ h is to fermentation ends.
The manufacture method of Aspergillus niger spores powder is: is washed out by aspergillus niger slant strains sterilized water, makes Aspergillus niger spores Concentration is 5 × 106The Aspergillus niger spores suspension of individual/mL, adds to Aspergillus niger spores suspension in activation medium, cultivates 25 for 36 DEG C Hour, then the Semen Cucurbitae shell powder with sterilizing mixes and mixes to obtain Aspergillus niger spores powder according to the part by weight of 1:2.Activation culture The raw material weight percentage ratio of base consists of: wheat bran 56%, Semen Cucurbitae grouts powder 10%, NaH2PO42%、MgSO41%、NaNO30.5%、 Yeast extract 0.5%, water 30%, pH is natural, 121 DEG C, sterilizing half an hour, be cooled to 35-40 DEG C.
Nutritional solution formula by weight percentage is: concentration of glucose 3%, potassium dihydrogen phosphate 1.6%, nicotinic acid 2.2 ‰, gallbladder Alkali 1.8 ‰, folic acid 2.23 ‰, VB22.8 ‰, water surplus.
(3) extraction of Semen Cucurbitae dregs of rice molecular peptide separates: the weight ratio that tunning is pressed 1:22 mixes with water, 58 DEG C of stirrings Soaking 5 hours, through filter press, filtrate is filtered through the daltonian hollow fiber column of trapped molecular weight 10000 and is retained, and obtains ultrafiltration Liquid, ultrafiltrate is through concentrating under reduced pressure, and sample adsorbs with macroporous resin column on the flow velocity of 2BV/h (macroporous resin D3520), then with 4BV/h flow velocity, uses 70% ethanol solution eluting instead when being colourless with 20% ethanol solution flushing to effluent, and 70% ethanol solution is washed De-elution speed is 2BV/h, collects the eluent of 70% ethanol solution eluting, after eluent concentrating under reduced pressure, is spray-dried, Semen Cucurbitae dregs of rice molecular peptide.The ventilation temperature being spray-dried is 210 DEG C, and leaving air temp 95 DEG C, feed rate is 30L/h.
The method of the present invention, the conversion ratio of Semen Cucurbitae dregs of rice molecular peptide reaches 75-85%, the nitrogen content of Semen Cucurbitae dregs of rice molecular peptide More than 95%.
Embodiment described above is the one preferably scheme of the present invention, not makees the present invention any pro forma Limit, on the premise of without departing from the technical scheme described in claim, also have other variant and remodeling.

Claims (10)

1. the method that Semen Cucurbitae dregs of rice molecular peptide prepared by mixed vaccine solid fermentation Semen Cucurbitae grouts, it is characterised in that include as Lower step:
(1) lactic acid bacteria fermentation: lactic acid bacteria and sterilized rice flour according to the weight ratio of 1:4-6 mix lactic acid bacteria rice flour mixes Thing, puts into mix homogeneously in fermentation medium by lactic acid bacteria rice flour mixture, and lactic acid bacteria rice flour amount of mixture is fermentation culture The 10-15% of basic weight amount, 35-40 DEG C of fermentation culture；
(2) fermentation of Aspergillus niger: when the pH of lactic acid bacteria fermentation cultivation to fermentation medium is down to 4.5-5.0, access aspergillus niger spore Sub-powder, Aspergillus niger spores powder consumption is the 15-18% of fermentation medium weight, and 32-35 DEG C of fermentation culture 70-72 hour must be fermented Product, the blowing air amount that fermentation of Aspergillus niger is cultivated first 24 hours is 4000-4500m3/ h, the logical sky of fermentation culture 25-49 hour Tolerance is 2500-3500m3/ h, when fermentation culture to 50h, adds fermentation medium weight 15-20% in fermentation medium The nutritional solution of weight, control blowing air amount is 900-1100m simultaneously3/ h is to fermentation ends；
(3) extraction of Semen Cucurbitae dregs of rice molecular peptide separates: the weight ratio that tunning is pressed 1:20-25 mixes with water, and 55-60 DEG C is stirred Mixing immersion 4-6 hour, through filter press, filtrate is filtered through the daltonian hollow fiber column of trapped molecular weight 10000 and is retained, Ultrafiltrate, ultrafiltrate is through concentrating under reduced pressure, and upper macroporous resin column is adsorbed, eluting, after eluent concentrating under reduced pressure, is spray-dried, obtains southern Melon seed dregs of rice molecular peptide.
Method the most according to claim 1, it is characterised in that described in step (1), fermentation medium is by weight percentage Meter composition is as follows: Semen Cucurbitae grouts powder 70%, Semen Cucurbitae shell powder 10%, water 20%；PH is natural, 121 DEG C, sterilizing half an hour, is cooled to 35-40℃。
Method the most according to claim 2, it is characterised in that described Semen Cucurbitae grouts powder is after Semen Cucurbitae grouts are pulverized , Semen Cucurbitae grouts powder size is at 40-60 mesh, and the size of Semen Cucurbitae shell powder is at 20-40 mesh.
Method the most according to claim 1 and 2, it is characterised in that in step (1), the manufacture method of lactic acid bacteria is: Adding 50-70kg whole milk powder and 45-55kg sucrose in 500L water, heated and stirred is dissolved, 115 DEG C, sterilizing 30-under 0.1MPa 60min, is cooled to 40-42 DEG C, is subsequently adding 5-10kg lactobacillus powder and stirs, and keeps the little fermentation culture of temperature 40-42 DEG C 8-12 hour.
Method the most according to claim 1 and 2, it is characterised in that the manufacture method of Aspergillus niger spores powder in step (2) For: being washed out by aspergillus niger slant strains sterilized water, making Aspergillus niger spores concentration is 2 × 106 -7×106The black fermented preparation of individual/mL Mould spore suspension, adds to Aspergillus niger spores suspension in activation medium, cultivates 24-26 hour, then with sterilizing for 35-38 DEG C Semen Cucurbitae shell powder mixing mix to obtain Aspergillus niger spores powder according to the part by weight of 1:1.5-2.5.
Method the most according to claim 5, it is characterised in that the raw material weight percentage ratio composition of described activation medium For: wheat bran 56%, Semen Cucurbitae grouts powder 10%, NaH2PO42%、MgSO41%、NaNO30.5%, yeast extract 0.5%, water 30%, pH is certainly So, 121 DEG C, sterilizing half an hour, be cooled to 35-40 DEG C.
Method the most according to claim 1 and 2, it is characterised in that step (2) Middle nutrition liquid is by weight percentage Formula is: concentration of glucose 3%, potassium dihydrogen phosphate 1.6%, nicotinic acid 2.2 ‰, choline 1.8 ‰, folic acid 2.23 ‰, VB22.8 ‰, Water surplus.
Method the most according to claim 1 and 2, it is characterised in that in step (3), the resin of macroporous resin column is macropore tree Fat D3520, macroporous resin column is adsorbed, the technological parameter of eluting is: sample, with macroporous resin column on the flow velocity of 2BV/h, is then used 20% ethanol solution flushing uses 70% ethanol solution eluting instead when being colourless to effluent, collects the eluting of 70% ethanol solution eluting Liquid.
Method the most according to claim 8, it is characterised in that the flow velocity that 20% ethanol solution rinses is 4BV/h, 70% ethanol The elution speed of eluant solution is 2BV/h.
Method the most according to claim 1 and 2, it is characterised in that the ventilation temperature being spray-dried in step (3) is 210 DEG C, leaving air temp 95 DEG C, feed rate is 30L/h.
CN201610706267.1A 2016-08-23 2016-08-23 Method for preparing pumpkin seed meal molecular peptide through solid state fermentation on pumpkin seed cake meal by virtue of mixed bacteria Pending CN106318999A (en)

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CN106916854A (en) * 2017-05-16 2017-07-04 湖南农业大学 A kind of tobacco black shank fermented liquid pathogenic activity composition and preparation method thereof
CN106987617A (en) * 2017-05-16 2017-07-28 湖南农业大学 A kind of application process of tobacco black shank fermented liquid pathogenic activity component
CN111700218A (en) * 2020-05-13 2020-09-25 中国农业科学院农产品加工研究所 Corn functional fungus grain and preparation method thereof

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106916854A (en) * 2017-05-16 2017-07-04 湖南农业大学 A kind of tobacco black shank fermented liquid pathogenic activity composition and preparation method thereof
CN106987617A (en) * 2017-05-16 2017-07-28 湖南农业大学 A kind of application process of tobacco black shank fermented liquid pathogenic activity component
CN106987617B (en) * 2017-05-16 2020-05-15 湖南农业大学 Application method of tobacco black shank bacterium fermentation broth pathogenic active component
CN106916854B (en) * 2017-05-16 2020-09-22 湖南农业大学 Tobacco black shank bacterium fermentation broth pathogenic active ingredient and preparation method thereof
CN111700218A (en) * 2020-05-13 2020-09-25 中国农业科学院农产品加工研究所 Corn functional fungus grain and preparation method thereof
CN111700218B (en) * 2020-05-13 2022-06-21 中国农业科学院农产品加工研究所 Corn functional fungus grain and preparation method thereof

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