CN104839445A - Production method of functional protein feed rich in two kinds of essential amino-acids - Google Patents
Production method of functional protein feed rich in two kinds of essential amino-acids Download PDFInfo
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Abstract
The invention discloses a production method of a functional protein feed rich in two kinds of essential amino-acids. The production method comprises the following steps: carrying out liquid fermentation on corynebacterium glutamicum, geotrichum candidum bacteria, lactobacillus plantarum and bacillus natto, mixing krausen liquid rich in metabolite and thallus to prepare krausen liquid, mixing the krausen liquid and a mixture of one or more of bean pulp, cottonseed meal, rapeseed meal and peanut meal according to a certain proportion, carrying out solid state fermentation so as to obtain a product, and finally carrying drying package on the product. The content of crude protein of obtained protein feed is higher than 50%, and the protein feed is rich in two kinds of essential amino-acids. The production method is simple in process, the krausen liquid is directly mixed with a solid state material for fermenting, probability of infectious microbe infection is reduced, energy consumption is low, and waste is not discharged. With adoption of fermentation, the content of antinutritional factors of plant meal raw materials is reduced, the contents of essential amino-acids and protein of the raw materials are improved, the economic additional value of agricultural and sideline products is greatly increased, the adding amount of essential amino-acids of an external source is reduced or replaced, and the feed cost is reduced.
Description
Technical field
The present invention relates to animal feed production technical field, especially relate to a kind of functional protein feed production method being rich in two kinds of essential amino acids.
Background technology
China's aquaculture is flourishing, and the main source of the annual a large amount of forage protein raw materials consumed is dregs of beans and fish meal.Problems such as although the crude protein content of these two kinds of products are higher, and fish meal exists quality heterogeneity, resource worsening shortages, though and meal expeller and mealsolvent cost is lower, but belong to vegetable protein, wherein containing a large amount of ANFs, affect the absorption and digestion of animal to feed.For addressing this problem, in recent years occurred utilizing lactic acid bacteria, bacillus, the mixed fungus fermentation such as saccharomycete and photosynthetic bacteria produces the technology of forage protein raw material.This type of technology utilizes the agricultural byproducts such as fermentation grouts, reaches elimination ANFs, increases protein content object, prepare protein feed method as national patent (publication No.: CN102488084A) discloses a kind of mixed aerobic bacteria fermented stalk grouts; National patent (publication No.: CN101449739) also discloses a kind of micro-ecology fermentation protein feedstuff and preparation method thereof.But common fermented feed production technology is difficult to the content and the Compositional balance problem that solve amino acid whose equilibrium problem, particularly essential amino acid (as arginine, lysine, threonine etc.) in protein feed usually.Current research shows, supplements the low-protein diet containing essential amino acid, can reduce feeding cost, reduces plant's environmental pollution, reduces cub diarrhea rate.But the amino acid used in current feed is nearly all first by biological fermentation process production sterling, then mixing is added in concentrate feed or complete diet pellet, has increased substantially feeding cost.
Use the microorganism such as bacillus, saccharomycete, lactic acid bacteria can the agricultural byproducts such as direct fermentation dregs of beans, although the ANFs in dregs of beans can be eliminated, improve digestibility, as national patent (publication No.: CN102406134A) just provides a kind of process for solid state fermentation of high digestibility fermented bean dregs, but the fermentation protein feedstuff that these methods are produced is difficult to improve the content of wherein essential amino acid.
Summary of the invention
The object of the present invention is to provide and a kind ofly utilize one or more mixed culture fermentations in dregs of beans, cotton dregs, rapeseed dregs and peanut meal to obtain the functional protein feed production method being rich in two kinds of essential amino acids.
For achieving the above object, the present invention can take following technical proposals:
The functional protein feed production method being rich in two kinds of essential amino acids of the present invention comprises the steps:
The first step, liquid fermentation
First seed culture and liquid fermentation is carried out in the mixing of 1:1 ratio by any two kinds that are numbered in CICC 20906, CICC 20191, CICC 20903, CICC 20887 and CICC 20,192 five kinds of Corynebacterium glutamicums; The Geotrichum being numbered CICC 1410 is carried out seed culture, is then linked into mixed fungus fermentation in Corynebacterium glutamicum zymotic fluid, obtain mixed fungus fermentation liquid for subsequent use;
Fermented separately by the Lactobacillus plantarum being numbered CICC20022 or CICC20038, zymotic fluid is for subsequent use;
Fermented separately by the bafillus natto being numbered CICC23413 or CICC20443, zymotic fluid is for subsequent use;
Second step, prepares fermentation liquid
By zymotic fluid obtained for the first step by Corynebacterium glutamicum and geotrichum candidum mixed fungus fermentation liquid 10 ~ 60 parts, Lactobacillus plantarum zymotic fluid 0 ~ 30 part or/and bacillus natto to ferment liquid 0 ~ 30 part mixing, with Ca (OH)
2adjust ph, to neutral, obtains fermentation liquid for subsequent use;
3rd step, solid state fermentation
By weight being mixed with 5 ~ 10 parts of molasses by one or more materials in 100 ~ 150 parts of dregs of beans, cotton dregs, rapeseed dregs and peanut meals, obtain solid state fermentation raw material; The fermentation liquid that second step is obtained in solid state fermentation raw material weight 50 ~ 100% ratio mix with fermentation raw material, carry out aerobic solid state fermentation, fermentation condition is: initial temperature 20 ~ 30 DEG C, process control temp 30 DEG C ~ 45 DEG C, and fermentation time is 48 ~ 72h;
4th step, drying and packaging
Fermentation ends, dries water content≤12% under 60 ~ 70 DEG C of conditions by fermentation materials, metering packing after pulverizing, can obtain the high protein feed being rich in two kinds of essential amino acids.
The preparation method of described first step Glutamic Acid bar bacterium and geotrichum candidum mixed fungus fermentation liquid is:
Any two kinds of Corynebacterium glutamicum kind SPSSs after using beef-protein medium activation are made suspension bacteria liquid, and be inoculated in liquid fermentation seed culture medium and cultivate, culture medium forms: glucose 100 ~ 120g/L, (NH
4)
2sO
420 ~ 30g/L, corn steep liquor 10 ~ 20g/L, KH
2pO
41 ~ 1.5g/L, MgSO
47H
2o 0.3 ~ 0.5 g/L, biotin 5 ~ 8 × 10
-5g/L, CaCO
310 ~ 30 g/L; PH7.0, sterilizing 20 min under 0.12 MPa; Condition of culture: tank pressure 0.03 ~ 0.1 Mpa, speed of agitator 170 ~ 230 r/min, ventilation 1:0.25 ~ 0.5, cultivation temperature 30 ~ 32 DEG C, fermentation time 24 ~ 48 h; Geotrichum SPSS is made suspension bacteria liquid, and when the acid production rate of Corynebacterium glutamicum zymotic fluid reaches more than 4.1%, access geotrichum candidum suspension bacteria liquid, continues cultivation and fermentation 10h.
In the described first step, the preparation method of Lactobacillus plantarum zymotic fluid is:
Solid MRS is used to cultivate Lactobacillus plantarum, Lactobacillus plantarum one-level solid seed SPSS is made suspension bacteria liquid, be inoculated in liquid fermentation medium, culture medium forms: glucose or molasses 20-50g/L, corn steep liquor 20 ~ 35 g/L, bitter salt 0.25 ~ 0.35 g/L, four hydrated manganese sulfate 0.01 ~ 0.015 g/L, seven ferric sulfate hydrate 0.01 ~ 0.015 g/L, sodium acetate 16 ~ 21 g/L, Tween-80 l mL/L; Condition of culture: tank pressure 0.03 ~ 0.1 Mpa quiescent culture, cultivation temperature 30 ~ 32 DEG C, fermentation time 18 ~ 22 h.
In the described first step, the preparation method of bacillus natto to ferment liquid is:
Bafillus natto kind SPSS after using beef-protein medium activation is made suspension bacteria liquid, be inoculated in liquid fermentation medium, culture medium forms: bean cake powder 1.0 ~ 3.0%, corn flour 1.0 ~ 3.0%, magnesium sulfate 0.2 ~ 0.6%, manganese sulfate 0.02 ~ 0.06%, sodium chloride 0.5 ~ 1.0%; Condition of culture: tank pressure 0.05 ~ 0.1 Mpa, speed of agitator 170 ~ 230 r/min, ventilation 1:0. 5 ~ 1.2, cultivation temperature 32 ~ 35 DEG C, fermentation time 18 ~ 24 h.
The invention has the advantages that technique is simple, by fermentation liquid, direct and solid material mixed culture fermentation, both make use of the metabolite of microorganism, had substantially reduced again the solid state fermentation time, reduced the probability contaminating miscellaneous bacteria in Solid Fermentation of Uncooked Material process; Energy consumption is low, no waste discharge in sweat; By fermentation, reduce ANFs content in plant waste matter class raw material, turn improve essential amino acid and protein content in raw material, greatly improve the economic value added of agricultural byproducts; In finished product feedstuff, essential amino acids content is high, can reduce or the addition of alternative external source essential amino acid, reduce feed cost.
Be employed herein five kinds of Corynebacterium glutamicums, the produced lysine of numbering CICC 20906, the produced arginine of numbering CICC 20191, the produced threonine of numbering CICC 20903, numbering CICC 20887 can produce valine, numbering CICC 20192 can produce tryptophan, by its any two kinds of combinations and geotrichum candidum, Lactobacillus plantarum and bafillus natto carry out seed culture and liquid fermentation respectively, the fermentation liquid obtained and dregs of beans, cotton dregs, one or more materials in rapeseed dregs and peanut meal are mixed thoroughly and are carried out solid state fermentation, obtain being rich in lysine, arginine, threonine, the high protein feed raw material of any two kinds of essential amino acids in valine and tryptophan, solve amino acid whose equilibrium problem in protein feed, particularly essential amino acid is (as arginine, lysine, threonine, valine and tryptophan etc.) content and Compositional balance problem.
Detailed description of the invention
Below in conjunction with embodiment, the invention will be further described.
Material proportion in following examples is mass percent.
The present invention bacterial classification Corynebacterium glutamicum used (
corynebacterium glutamicum, CICC 20906, CICC 20191, CICC 20903, CICC 20887, CICC 20192), Geotrichum (
candida utilis, CICC 1410), Lactobacillus plantarum (
lactobacillus plantarum, CICC20022, CICC20038), bafillus natto (
bacillus natto, CICC23413, CICC20443) all purchased from Chinese industrial Microbiological Culture Collection administrative center.
embodiment 1
Of the present inventionly be rich in lysine, the functional protein feed production method of arginine two kinds of essential amino acids comprises the steps:
The first step, liquid fermentation
On beef extract-peptone two kind of Corynebacterium glutamicum (being numbered CICC 20906 and CICC 20191) is made suspension bacteria liquid with 200ml SPSS, after the mixing of 1:1 ratio, be inoculated in liquid fermentation medium and cultivate (liquid amount 60%), culture medium forms: glucose 120g/L, (NH
4)
2sO
430g/L, corn steep liquor 20g/L, KH
2pO
41.5g/L, MgSO
47H
2o0.5 g/L, biotin 8 × 10
-5g/L, CaCO
330 g/L; PH7.0, sterilizing 20 min under 0.12 MPa; Condition of culture: tank pressure 0.1 Mpa, speed of agitator 230 r/min, ventilation 1:0.25, cultivation temperature 30 DEG C, fermentation time 48 h; Geotrichum (being numbered CICC 1410) on YPD culture medium is made suspension bacteria liquid with 200ml SPSS, and when the acid production rate of Corynebacterium glutamicum zymotic fluid reaches more than 4.1%, access Geotrichum suspension bacteria liquid, continues cultivation and fermentation 10h;
Lactobacillus plantarum (being numbered CICC20022) on inclined-plane is inoculated in the flat bottle of 500mL eggplant shape filling MRS culture medium, Lactobacillus plantarum one-level solid seed 200mL SPSS is made suspension bacteria liquid, (liquid amount 60%) is cultivated in fermentation tank, culture medium forms: glucose or molasses 20-50g/L, corn steep liquor 35 g/L, bitter salt 0.35 g/L, four hydrated manganese sulfate 0.015 g/L, seven ferric sulfate hydrate 0.015 g/L, sodium acetate 21 g/L, Tween-80 lmL/L; Condition of culture: tank pressure 0.1 Mpa quiescent culture, cultivation temperature 30 DEG C, fermentation time 22 h.
Second step, prepares fermentation liquid
The zymotic fluid that the first step is obtained mixes with Lactobacillus plantarum zymotic fluid 10 parts by Corynebacterium glutamicum and geotrichum candidum mixed fungus fermentation liquid 20 parts, with Ca (OH)
2adjust ph to 7.0, obtains fermentation liquid for subsequent use;
3rd step, solid state fermentation
By weight being mixed with 5 parts of molasses by 100 parts of dehulled soybean meals, obtain solid state fermentation raw material; By second step prepare fermentation liquid in solid state fermentation raw material weight 60% ratio mix with solid state fermentation raw material, fermentation tower carries out aerobic solid state fermentation, fermentation condition: initial temperature 25 DEG C, process control temp 45 DEG C, fermentation time is 60h;
4th step, drying and packaging
Fermentation ends, is sent to fermentation materials on drier, uses hot blast, under 65 DEG C of conditions, dry water content≤12%, metering packing after pulverizing, and obtaining being rich in two kinds must amino acid whose high protein feed raw material.
Measure according to national standard (GB/T 19541 and GB/T 18296), the nutritive index of the fermented bean dregs that the inventive method obtains and normal meal, normal fermentation dregs of beans contrasts, and result is as follows:
Can find out from upper table, in the fermented bean dregs that the inventive method obtains, crude protein content reaches 51.3%, and the arginine content in essential amino acid can reach 4.09%, and lysine content can reach 3.77%.In General Promotion of the present invention feed two kinds must amino acid whose content, there is no the appearance of similar technique at present, simultaneously the good palatability of feed, be easy to animal and absorb and utilize.
embodiment 2
Of the present inventionly be rich in threonine, the functional protein feed production method of arginine two kinds of essential amino acids comprises the steps:
The first step, liquid fermentation
Two kinds of Corynebacterium glutamicums (being numbered CICC 20903 and CICC 20191) mix the preparation method of bacterium liquid with embodiment 1 with Geotrichum (being numbered CICC 1410);
Get bafillus natto (being numbered CICC23413) inclined-plane seed to inoculate in the 500ml eggplant-shape bottle filling peptone beef extract culture medium, bafillus natto one-level solid seed 200ml SPSS is made suspension bacteria liquid, be inoculated into (seed tank culture base liquid amount 60%) fermented and cultured in 2m3 fermentation tank, culture medium forms: bean cake powder 2.0%, corn flour 1.5%, magnesium sulfate 0.4%, manganese sulfate 0.05%, sodium chloride 1.0%; Condition of culture: tank pressure 0.05 Mpa, speed of agitator 190 r/min, ventilation 1:0.8, cultivation temperature 35 DEG C, fermentation time 20 h.
Second step, prepares fermentation liquid
The zymotic fluid that the first step is obtained mixes with bacillus natto to ferment liquid 10 parts by Corynebacterium glutamicum and geotrichum candidum mixed fungus fermentation liquid 30 parts, with Ca (OH)
2adjust ph to 7.0, obtains fermentation liquid for subsequent use;
3rd step, solid state fermentation
By weight being mixed with 10 parts of molasses by 120 parts of dehulled soybean meals, obtain solid state fermentation raw material; By second step prepare fermentation liquid in solid state fermentation raw material weight 80% ratio mix with solid state fermentation raw material, fermentation tower carries out aerobic solid state fermentation, fermentation condition: initial temperature 25 DEG C, process control temp 45 DEG C, fermentation time is 60h;
4th step, drying and packaging
Fermentation ends, is sent to fermentation materials on drier, uses hot blast, under 65 DEG C of conditions, dry water content≤12%, metering packing after pulverizing, and obtaining being rich in two kinds must amino acid whose high protein feed raw material.
Measure according to national standard (GB/T 19541 and GB/T 18296), the nutritive index of the fermented bean dregs that the inventive method obtains and normal meal, normal fermentation dregs of beans contrasts, and result is as follows:
Can find out from upper table, in the fermented bean dregs that the inventive method obtains, crude protein content reaches 51.8%, and the arginine content in essential amino acid can reach 4.17%, and threonine content can reach 2.38%.In General Promotion of the present invention feed two kinds must amino acid whose content, there is no the appearance of similar technique at present, simultaneously the good palatability of feed, be easy to animal and absorb and utilize.
embodiment 3
Of the present inventionly be rich in threonine, the functional protein feed production method of lysine two kinds of essential amino acids comprises the steps:
The first step, liquid fermentation
Two kinds of Corynebacterium glutamicums (being numbered CICC 20903 and CICC 20906) and with the preparation method of the mixed bacterium liquid of Geotrichum (being numbered CICC 1410) with embodiment 1; Lactobacillus plantarum zymotic fluid production method is identical with the step (1) in embodiment 1; Bacillus natto to ferment liquid production method is identical with the step (1) in embodiment 2;
Second step, prepares fermentation liquid
By zymotic fluid obtained for the first step by Corynebacterium glutamicum and Geotrichum mixed fungus fermentation liquid 40 parts, Lactobacillus plantarum zymotic fluid 20 parts, bacillus natto to ferment liquid 20 parts, mixes, obtains fermentation liquid.
3rd step, solid state fermentation
By weight, dehulled soybean meal 150 parts and 10 parts of molasses are mixed to get solid state fermentation raw material; By second step prepare fermentation liquid in solid state fermentation raw material weight 80% ratio mix with solid state fermentation raw material, fermentation tower carries out aerobic solid state fermentation; Solid state fermentation conditions is identical with embodiment 1;
4th step, drying and packaging
Fermentation ends, is sent to fermentation materials on drier, uses hot blast, under 65 DEG C of conditions, dry water content≤12%, metering packing after pulverizing, and obtaining being rich in two kinds must amino acid whose high protein feed raw material.
The fermented bean dregs protein raw materials that production method of the present invention obtains after measured (assay method is with embodiment 1,2) sees the following form with the comparing result of dregs of beans, normal fermentation dregs of beans protein raw materials nutritive index.
Can find out from upper table, in the fermented bean dregs that the inventive method obtains, crude protein content reaches 51.9%, and the lysine content in essential amino acid can reach 3.65%, and threonine content can reach 2.34%.In General Promotion of the present invention feed two kinds must amino acid whose content, there is no the appearance of similar technique at present, simultaneously the good palatability of feed, be easy to animal and absorb and utilize.
In above-described embodiment, solid state fermentation raw material also can use one or more mixtures in dregs of beans, cotton dregs, rapeseed dregs and peanut meal to replace.
Claims (4)
1. be rich in a functional protein feed production method for two kinds of essential amino acids, it is characterized in that: comprise the steps:
The first step, liquid fermentation
First seed culture and liquid fermentation is carried out in after the mixing of 1:1 ratio by any two kinds that are numbered in CICC 20906, CICC 20191, CICC 20903, CICC 20887 and CICC 20,192 five kinds of Corynebacterium glutamicums; The Geotrichum being numbered CICC 1410 is carried out seed culture, is then linked into mixed fungus fermentation in Corynebacterium glutamicum zymotic fluid, obtain mixed fungus fermentation liquid for subsequent use;
Fermented separately by the Lactobacillus plantarum being numbered CICC20022 or CICC20038, zymotic fluid is for subsequent use;
Fermented separately by the bafillus natto being numbered CICC23413 or CICC20443, zymotic fluid is for subsequent use;
Second step, prepares fermentation liquid
By zymotic fluid obtained for the first step by Corynebacterium glutamicum and geotrichum candidum mixed fungus fermentation liquid 10 ~ 60 parts, Lactobacillus plantarum zymotic fluid 0 ~ 30 part or/and bacillus natto to ferment liquid 0 ~ 30 part mixing, with Ca (OH)
2adjust ph, to neutral, obtains fermentation liquid for subsequent use;
3rd step, solid state fermentation
By weight one or more materials in 100 ~ 150 parts of dregs of beans, cotton dregs, rapeseed dregs and peanut meals are mixed with 5 ~ 10 parts of molasses, obtain solid state fermentation raw material, the fermentation liquid obtained by second step mixes with fermentation raw material in the ratio of solid state fermentation raw material weight 50 ~ 100%, carry out aerobic solid state fermentation, fermentation condition is: initial temperature 20 ~ 30 DEG C, process control temp 30 DEG C ~ 45 DEG C, fermentation time is 48 ~ 72h;
4th step, drying and packaging
Fermentation ends, dries water content≤12% under 60 ~ 70 DEG C of conditions by fermentation materials, metering packing after pulverizing, can obtain the high protein feed being rich in two kinds of essential amino acids.
2. the functional protein feed production method being rich in two kinds of essential amino acids according to claim 1, is characterized in that: the preparation method of described first step Glutamic Acid bar bacterium and geotrichum candidum mixed fungus fermentation liquid is:
Any two kinds of Corynebacterium glutamicum kind SPSSs after using beef-protein medium activation are made suspension bacteria liquid, and be inoculated in liquid fermentation seed culture medium and cultivate, culture medium forms: glucose 100 ~ 120g/L, (NH
4)
2sO
420 ~ 30g/L, corn steep liquor 10 ~ 20g/L, KH
2pO
41 ~ 1.5g/L, MgSO
47H
2o 0.3 ~ 0.5 g/L, biotin 5 ~ 8 × 10
-5g/L, CaCO
310 ~ 30 g/L; PH7.0, sterilizing 20 min under 0.12 MPa; Condition of culture: tank pressure 0.03 ~ 0.1 Mpa, speed of agitator 170 ~ 230 r/min, ventilation 1:0.25 ~ 0.5, cultivation temperature 30 ~ 32 DEG C, fermentation time 24 ~ 48 h; Geotrichum SPSS is made suspension bacteria liquid, and when the acid production rate of Corynebacterium glutamicum zymotic fluid reaches more than 4.1%, access geotrichum candidum suspension bacteria liquid, continues cultivation and fermentation 10h.
3. the functional protein feed production method being rich in two kinds of essential amino acids according to claim 1, is characterized in that: in the described first step, the preparation method of Lactobacillus plantarum zymotic fluid is:
Solid MRS is used to cultivate Lactobacillus plantarum, Lactobacillus plantarum one-level solid seed SPSS is made suspension bacteria liquid, be inoculated in liquid fermentation medium, culture medium forms: glucose or molasses 20-50g/L, corn steep liquor 20 ~ 35 g/L, bitter salt 0.25 ~ 0.35 g/L, four hydrated manganese sulfate 0.01 ~ 0.015 g/L, seven ferric sulfate hydrate 0.01 ~ 0.015 g/L, sodium acetate 16 ~ 21 g/L, Tween-80 l mL/L; Condition of culture: tank pressure 0.03 ~ 0.1 Mpa quiescent culture, cultivation temperature 30 ~ 32 DEG C, fermentation time 18 ~ 22 h.
4. the functional protein feed production method being rich in two kinds of essential amino acids according to claim 1, is characterized in that: in the described first step, the preparation method of bacillus natto to ferment liquid is:
Bafillus natto kind SPSS after using beef-protein medium activation is made suspension bacteria liquid, be inoculated in liquid fermentation medium, culture medium forms: bean cake powder 1.0 ~ 3.0%, corn flour 1.0 ~ 3.0%, magnesium sulfate 0.2 ~ 0.6%, manganese sulfate 0.02 ~ 0.06%, sodium chloride 0.5 ~ 1.0%; Condition of culture: tank pressure 0.05 ~ 0.1 Mpa, speed of agitator 170 ~ 230 r/min, ventilation 1:0. 5 ~ 1.2, cultivation temperature 32 ~ 35 DEG C, fermentation time 18 ~ 24 h.
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