CN110583855A

CN110583855A - Preparation method of high tryptophan fermented feed

Info

Publication number: CN110583855A
Application number: CN201911009194.0A
Authority: CN
Inventors: 符秀迪; 程勐万里; 孟龙; 薛芹
Original assignee: Hefei Wuliangye Thai Biotechnology Co Ltd
Current assignee: Hefei Wuliangye Thai Biotechnology Co Ltd
Priority date: 2019-10-23
Filing date: 2019-10-23
Publication date: 2019-12-20

Abstract

The invention discloses a preparation method of high tryptophan fermented feed, which comprises the following steps: crushing a starch-containing product, adding water for size mixing, adding amylase for enzymolysis, liquefaction and saccharification to obtain a saccharified liquid with residues, adding a nutrient solution required by microbial growth, performing high-temperature sterilization, cooling to a suitable temperature for microbial growth, directly inoculating a tryptophan-producing strain, introducing sterile air for fermentation for a plurality of hours, and performing solid-liquid separation to obtain a tryptophan thallus wet base and a tryptophan fermentation liquid; regulating the pH value of the tryptophan fermentation liquid to an isoelectric point, and then carrying out reduced pressure evaporation and concentration to prepare a fermentation source tryptophan concentrated solution; mixing one or more of the sugar residue wet base and the tryptophan thallus wet base with the composite probiotic bacteria liquid and the composite enzyme, and performing enzymolysis fermentation to obtain a zymolysis residue meal wet base; and (3) preparing the high tryptophan fermented feed. The tryptophan fermented feed prepared by the invention has the advantages of low production equipment investment, no need of refining and extraction, high product yield, no wastewater generation and low production cost.

Description

Preparation method of high tryptophan fermented feed

Technical Field

The invention belongs to the technical field of feeds, enzyme preparations and microorganisms, and particularly relates to a preparation method of a high tryptophan fermentation feed.

Background

China is a big population country and also the first pig-raising country in the world. In 2017, the livestock production value in China reaches 3.6 trillion yuan, live pigs are bred by 6.89 trillion heads, the breeding stock of sows is 4200 trillion heads, and the pork yield is 5487 trillion tons. According to the statistical data of Ministry of agriculture, the national commercial feed yield in 2017 reaches 2.05 hundred million tons. Wherein, the complete feed is 1.79 million tons, the concentrated feed is 2151 million tons, the premix compound is 690 million tons, and comprises 8974 million tons of pig feed output, 9000 million tons of poultry feed output, 5300 million tons of aquatic feed, 850 million tons of cattle and sheep feed, 350 million tons of feed such as rabbit, racoon dog and pet animals, and 500 more than ten million tons of feed for the national sow. The national compound feed yield is expected to reach more than 2.2 hundred million tons in 2020, wherein the feed for sows reaches more than 800 ten thousand tons. The animal protein feed is relatively deficient in resources, and the content of essential amino acid in the vegetable protein is relatively low.

Tryptophan is a neutral aromatic amino acid containing an indole group, is one of essential amino acids in animal life activities, plays an important role in growth, development and metabolism of animals, and is called as a second essential amino acid. Numerous studies have been carried out on the nutritional metabolism of tryptophan, and the absence of tryptophan results in decreased feed intake, retarded growth, and rough hair cover in animals. Tryptophan cannot be synthesized in animal bodies, and tryptophan in plant feed cannot meet the requirements of livestock and poultry generally. In recent years, synthetic lysine and methionine have been used in large amounts in compound feeds, and the importance of tryptophan in food and feed has been clearly shown. Meanwhile, the lack of tryptophan affects the utilization rate of other amino acids, reduces the effective utilization rate of the whole protein, and causes that animals grow slowly and have high feed-meat ratio.

The traditional tryptophan production method needs food-grade glucose or glucose syrup, tryptophan fermentation liquor is obtained through fermentation, and tryptophan products are obtained through processes of thallus filtration, decoloration, ion-exchange crystallization and the like, so that a large amount of waste water is generated, the product yield is only less than 80%, the industrial production cost is high, and the adding cost in the feed is directly high.

Disclosure of Invention

The invention aims to solve the defects of insufficient essential amino acid resources and high industrial production cost, and provides the high tryptophan fermented feed which is rich in nutritional ingredients such as high tryptophan, small peptides, oligosaccharides, lactic acid, probiotics and the like and has the characteristics of growth promotion, good palatability, balanced amino acids, high nutrition, easy digestion, animal immunity enhancement and the like.

The technical scheme adopted by the invention is as follows:

a preparation method of a high tryptophan fermentation feed comprises the following steps:

s1, preparation of tryptophan fermentation liquor and tryptophan thallus wet basis:

crushing a starch-containing product, adding water for size mixing, adding amylase for enzymolysis, liquefaction and saccharification to obtain a saccharified liquid with residues, adding a nutrient solution required by microbial growth, performing high-temperature sterilization, cooling to a suitable temperature for microbial growth, directly inoculating a tryptophan-producing strain, introducing sterile air for fermentation for a plurality of hours, and performing solid-liquid separation to obtain a tryptophan thallus wet base and a tryptophan fermentation liquid;

s2, preparation of fermentation source tryptophan concentrated solution:

regulating the pH value of the tryptophan fermentation liquid to an isoelectric point, and then carrying out reduced pressure evaporation and concentration to prepare a fermentation source tryptophan concentrated solution;

s3, preparation of fermentation source tryptophan concentrated solution:

mixing one or more of the sugar residue wet base and the tryptophan thallus wet base with the composite probiotic bacteria liquid and the composite enzyme, and performing enzymolysis fermentation to obtain a zymolysis residue meal wet base;

s4, preparing the high tryptophan fermented feed:

mixing the fermentation source tryptophan concentrated solution and the glycolysis residue meal wet base, directly producing the high tryptophan fermentation feed wet base, or mixing and drying, or respectively drying and mixing to prepare a high tryptophan fermentation feed dry base;

preferably, in step S1, the tryptophan fermentation broth and the wet tryptophan biomass can be prepared by the following method: preparing tryptophan fermentation liquor and tryptophan thallus wet base or crushing starch-containing products, adding water for size mixing, adding amylase for enzymolysis, liquefying and saccharifying, and performing solid-liquid separation to obtain sugar residue wet base and residue-free saccharified liquid; adding nutrient solution required by microorganisms into the residue-free saccharified solution, sterilizing at high temperature, cooling to a suitable temperature required by the growth of the microorganisms, inoculating a tryptophan-producing strain, introducing sterile air, fermenting for several hours, and directly preparing tryptophan fermentation liquor.

Preferably, in step S3, the preparation of the wet base of the glycolysis residue meal can also adopt the following method: mixing one or more of the sugar residue wet base and the tryptophan thallus wet base with feed raw materials, clean water, composite probiotic bacteria liquid and composite enzyme, and performing enzymolysis and fermentation to obtain the glycolysis residue meal wet base.

Preferably, in step S4, the high tryptophan fermented feed can be prepared by the following method: mixing the fermentation source tryptophan concentrated solution, the fermented dreg.

Further, the nutrient solution required by the microorganism is one or more of yeast extract, yeast powder, peptone, beef extract, corn steep liquor, ammonium sulfate, magnesium sulfate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, vitamin B1 and biotin;

further, the feed raw material is one or more of bean pulp, cotton pulp, rapeseed pulp, peanut pulp, fish meal, blood meal, plasma protein, blood cell protein, milk powder, lactose, whey powder, wheat gluten powder, corn protein powder, rice protein powder, DDGS, soybean, pea, red bean, corn, rice, broken rice, wheat, bran, rice bran, flour, oat, barley, sweet potato, soybean protein isolate, soybean protein concentrate, pasture meal, Chinese herbal medicines, fruit and vegetable, sugar molasses syrup and byproducts of the food processing industry, or the puffed products of the feed raw materials;

further, the commercial non-fermentation source tryptophan for feed is one or more of non-fermentation tryptophan in the feed additive catalogue of the national ministry of agriculture;

further, the probiotic strain liquid is one or more of bacillus coagulans, bacillus subtilis, bacillus licheniformis, saccharomyces cerevisiae, aroma-producing yeast, lactobacillus acidophilus, bifidobacterium, enterococcus faecium, pediococcus acidilactici, lactobacillus plantarum and lactobacillus delbrueckii;

further, the complex enzyme is one or more of high-temperature alpha amylase, saccharifying enzyme, fungal enzyme, beta amylase, mannase, xylanase, glucanase, pectinase and protease.

Preferably, the nutrient solution required by the microorganism is yeast extract, peptone, ammonium sulfate, magnesium sulfate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, vitamin B1 and biotin;

preferably, the feed raw material is one or more of soybean meal, cottonseed meal, corn gluten meal, rice protein meal, DDGS, bran, rice bran, pasture grass meal, Chinese herbal medicines, fruits and vegetables, sugar molasses syrup and byproducts of the food processing industry, or puffed products of the feed raw material;

preferably, the commercial feed non-fermentation source tryptophan is one or more of synthetic pharmaceutical grade tryptophan, synthetic food grade tryptophan, industrial grade tryptophan, tryptophan additives, tryptophan derivatives;

preferably, the probiotic bacteria liquid is one or more of bacillus coagulans, bacillus subtilis, saccharomyces cerevisiae, lactobacillus acidophilus, bifidobacterium, enterococcus faecium, pediococcus acidilactici, lactobacillus plantarum and lactobacillus delbrueckii;

preferably, the complex enzyme is one or more of mannanase, xylanase, glucanase, pectinase and protease.

Further, the conditions for preparing the fermentation source tryptophan concentrate in step S2 are as follows:

(1) the microbial nutrient solution comprises the following components in percentage by weight: in the saccharified liquid with slag or saccharified liquid: water: yeast extract: peptone: ammonium sulfate: dipotassium hydrogen phosphate: potassium dihydrogen phosphate: magnesium sulfate: vitamin B1: the weight ratio of biotin is 100:0-100:0.01-2:0.01-4:0.01-1.5:0.01-1:0.01-1:0.01-0.5:0.00000001-0.1: 0.000000001-0.1;

(2) the sterilization and cooling conditions of the mixed microbial nutrient solution are as follows: sterilizing at 80-150 deg.C for 1-100 min, and cooling to 10-40 deg.C;

(3) when preparing the tryptophan fermentation liquor: fermentation temperature of tryptophan-producing recombinant escherichia coli strain: 10-40 ℃; fermentation pH: 5.0-7.0; stirring at 20-1000r/min for aerobic fermentation; the aeration ratio is 1:0.1-3 v/v; fermentation time: 6-216 hours;

(4) and after the pH value of the tryptophan fermentation liquid is adjusted to 1.0-7.0, the tryptophan fermentation liquid is directly subjected to reduced pressure evaporation concentration, and the concentration of a dry substance is 10-80%.

Preferably, the conditions for preparing the fermentation source tryptophan concentrate in the step S2 are as follows:

(1) the microbial nutrient solution comprises the following components in percentage by weight: in the saccharified liquid with slag or saccharified liquid: water: yeast extract: peptone: ammonium sulfate: dipotassium hydrogen phosphate: potassium dihydrogen phosphate: magnesium sulfate: vitamin B1: the weight ratio of biotin is 100:80-100:0.8-1:1.5-2: 0.5-0.6: 0.4-0.5: 0.4-0.5:0.1-0.2:0.000003-0.00001: 0.0000003-0.000001;

(2) the sterilization and cooling conditions of the mixed microbial nutrient solution are as follows: sterilizing at the high temperature of 115 ℃ and 125 ℃ for 20-30 minutes, and cooling to 35-40 ℃;

(3) and when preparing tryptophan fermentation liquor: fermentation temperature of tryptophan-producing recombinant escherichia coli strain: 35-38 ℃; fermentation pH: 6.4-6.8; stirring for 300-400r/min in aerobic fermentation; the aeration ratio is 1:1-1.2 v/v; fermentation time: 38-42 hours;

(4) and after the pH value of the tryptophan fermentation liquid is adjusted to 5.5-6.0, the tryptophan fermentation liquid is directly subjected to reduced pressure evaporation concentration, and the concentration of a dry substance is 50-55%.

Further, the preparation conditions of the wet base of the glycolysis residue meal in the step S3 are as follows:

(1) the weight ratio of the sugar residue wet basis to the tryptophan thallus wet basis to the feed raw materials, the clean water, the composite probiotic bacteria liquid and the composite enzyme is 100:0 to 500: 0-3000: 0 to 2000: 0.5-50: 0.0001 to 10;

(2) and lactobacillus strain liquid in the composite probiotic strain liquid: the yeast strain liquid: the viable bacteria number ratio of the bacillus licheniformis strain liquid is 100:0 to 200: 0 to 100 parts;

(3) commercial proteases in complex enzymes: xylanase: glucanase: mannanase: the weight ratio of the pectinase is 100:0 to 1000: 0 to 1000: 0 to 1000: 0 to 1000;

(4) and enzymolysis fermentation temperature: 10-60 ℃, initial pH of enzymolysis and fermentation: 4-7, performing anaerobic enzymatic hydrolysis fermentation for 12-240 hours after the first aerobic enzymatic hydrolysis fermentation.

Preferably, the preparation conditions of the wet base of the glycolysis residue meal in the step S3 are as follows:

(1) the weight ratio of the sugar residue wet base, the tryptophan thallus wet base to the feed raw materials, the clean water, the composite probiotic bacteria liquid and the composite enzyme is 100: 0-80: 200-800: 0 to 200: 10-20: 0.005 to 2;

(2) and lactobacillus strain liquid in the composite probiotic strain liquid: the yeast strain liquid: the viable bacteria number ratio of the bacillus licheniformis strain liquid is 100: 10-30: 5-10;

(3) commercial proteases in complex enzymes: xylanase: glucanase: mannanase: the weight ratio of the pectinase is 100: 60-80: 60-80: 60-80: 30-50;

(4) and enzymolysis fermentation temperature: 30-40 ℃, initial pH of enzymolysis and fermentation: 6-7, carrying out anaerobic enzymatic hydrolysis fermentation for 72-96 hours after the first aerobic enzymatic hydrolysis fermentation.

Further, the preparation conditions of the high tryptophan fermented feed in the step S4 are as follows:

the weight ratio of the fermentation source tryptophan concentrated solution, the fermented dreg wet base to the commercial non-fermentation source tryptophan is 100:0 to 2000: 0 to 300.

Preferably, the preparation conditions of the high tryptophan fermented feed in the step S4 are as follows:

the weight ratio of the fermentation source tryptophan concentrated solution, the fermented dreg wet base to the commercial non-fermentation source tryptophan is 100: 200-800: 20 to 40.

The invention has the beneficial effects that:

(1) the high tryptophan fermentation feed prepared by the invention uses the solid fermentation wet material as the carrier to adsorb the fermentation liquor, thereby reducing the drying difficulty of the tryptophan fermentation liquor, improving the yield, leading the yield of the main component tryptophan to reach 100 percent, not generating waste water, having less investment on production equipment and low production cost.

(2) The high tryptophan fermented feed prepared by the invention is fermented and enzymolyzed by a plurality of probiotics together, is rich in tryptophan, also contains nutrients such as small peptides, lactic acid, oligosaccharides and probiotics, and has the characteristics of growth promotion, good palatability, balanced amino acids, high nutrition, easy digestion, animal immunity enhancement and the like.

(3) The high tryptophan fermented feed prepared by the invention can obviously improve the production performance of weaned piglets, can obviously improve the production performance of broiler chickens and can obviously improve the egg laying performance of laying hens.

Detailed Description

Example 1

100kg of corn is crushed, 200kg of water is added for size mixing, and then liquefying enzyme and saccharifying enzyme are added for enzymolysis, so as to prepare 300kg of saccharified liquid with dregs. 258.4kg of clean water, 3kg of yeast extract, 6kg of peptone, 3kg of ammonium sulfate, 1.8kg of dipotassium hydrogen phosphate, 1.8kg of potassium dihydrogen phosphate, 1kg of magnesium sulfate and vitamin B were added to 300kg of the saccharified liquid with dregs₁30g of biotin and 3g of biotin, sterilizing for 30 minutes at the high temperature of 121 ℃, cooling to 36 ℃, inoculating 25kg of tryptophan-producing recombinant escherichia coli strain liquid, and stirring for 4 days under the conditions of 35-37 ℃ and fermentation pH6.5And (3) introducing sterile air at a speed of 00r/min for 0.72m3/h, and carrying out aerobic fermentation for 40 hours to obtain 600kg of crude tryptophan fermentation liquor. Solid-liquid separation to obtain 60kg of tryptophan thallus wet base and 540kg of tryptophan fermentation liquor.

S2, preparation of tryptophan concentrate:

540kg of tryptophan fermentation liquid is adjusted to pH 5.89, and 118kg of tryptophan concentrated liquid is prepared after direct reduced pressure evaporation and concentration.

S3, preparation of fermentation source tryptophan concentrated solution:

adding 15kg of soybean meal, 15kg of corn protein powder, 15kg of rice protein powder, 15kg of fruit and vegetable residues, 15kg of molasses, 50kg of clean water, 12kg of composite probiotic strain liquid and 0.08kg of composite enzyme into 60kg of tryptophan strain wet base, mixing, and performing pH value regulation at the temperature of 30-40 ℃ and at the initial pH value: under the condition of 6-7, carrying out anaerobic fermentation for 96 hours after first oxygenation, and preparing 197kg of glycolysis residue meal wet base.

S4, preparing the high tryptophan fermented feed:

after 118kg of tryptophan concentrated solution and 197kg of wet substrate of glycolysis residue meal are mixed, 315kg of wet substrate of high tryptophan fermentation feed with water content of 45-50% is directly produced.

Example 2

200kg of corn is crushed, 400kg of water is added for size mixing, and then liquefying enzyme and saccharifying enzyme are added for enzymolysis, so as to prepare 600kg of saccharified liquid with dregs. The 180 sugar residue wet basis and 420kg of the dreg-free saccharified liquid are obtained by solid-liquid separation, 361kg of clean water, 4.2kg of yeast extract, 8.4kg of peptone, 4.2kg of ammonium sulfate, 2.5kg of dipotassium hydrogen phosphate, 2.5kg of monopotassium phosphate, 1.4kg of magnesium sulfate, vitamin B and the like are added into the 420kg of the dreg-free saccharified liquid₁42g and 4.2g of biotin, sterilizing at the high temperature of 121 ℃ for 30 minutes, cooling to 36 ℃, inoculating 50kg of tryptophan-producing recombinant escherichia coli strain liquid, stirring at the temperature of 35-37 ℃ and under the condition of fermentation pH6.5, introducing sterile air of 1.0m3/h, and carrying out aerobic fermentation for 40 hours to obtain 850kg of tryptophan fermentation liquid.

S2, preparation of tryptophan concentrate:

850kg of tryptophan fermentation liquor is adjusted to pH 5.89, and is directly evaporated and concentrated under reduced pressure to prepare 175kg of tryptophan concentrated liquor.

S3, preparation of fermentation source tryptophan concentrated solution:

adding 45kg of bean pulp, 45kg of corn protein powder, 45kg of rice protein powder, 45kg of water, fruit, vegetable and residue, 150kg of clean water, 36kg of composite probiotic strain liquid and 0.2kg of composite enzyme into 180kg of tryptophan thallus wet base, mixing, and performing pH value regulation at the temperature of 30-40 ℃ and the initial pH value: and under the condition of 6-7, carrying out anaerobic fermentation for 96 hours after first oxygenation to obtain 546kg of fermentation residue meal wet base.

S4, preparing the high tryptophan fermented feed:

after 175kg of tryptophan concentrated solution and 546kg of wet substrate of the glycolysis residue meal are mixed, 721kg of wet substrate of the high tryptophan fermentation feed with the water content of 45-50% is directly produced.

Example 3

100kg of corn is crushed, 200kg of water is added for size mixing, and then liquefying enzyme and saccharifying enzyme are added for enzymolysis, so as to prepare 300kg of saccharified liquid with dregs. 258.4kg of clean water, 3kg of yeast extract, 6kg of peptone, 3kg of ammonium sulfate, 1.8kg of dipotassium hydrogen phosphate, 1.8kg of potassium dihydrogen phosphate, 1kg of magnesium sulfate and vitamin B were added to 300kg of the saccharified liquid with dregs₁30g and 3g of biotin, sterilizing at the high temperature of 121 ℃ for 30 minutes, cooling to 36 ℃, inoculating 25kg of recombinant escherichia coli strain liquid producing tryptophan, stirring at the temperature of 35-37 ℃ and under the condition of fermentation pH6.5, introducing sterile air of 0.72m3/h, and carrying out aerobic fermentation for 40 hours to prepare 600kg of crude tryptophan fermentation liquid. Solid-liquid separation to obtain 60kg of tryptophan thallus wet base and 540kg of tryptophan fermentation liquor.

S2, preparation of tryptophan concentrate:

S3, preparation of fermentation source tryptophan concentrated solution:

S4, preparing the high tryptophan fermented feed:

118kg of tryptophan concentrated solution and 197kg of wet substrate of glycolysis residue meal are mixed and dried to prepare 170kg of high tryptophan fermented feed dry substrate.

Example 4

S2, preparation of tryptophan concentrate:

S3, preparation of fermentation source tryptophan concentrated solution:

S4, preparing the high tryptophan fermented feed:

175kg of tryptophan concentrated solution and 546kg of fermented dreg wet base are mixed and dried to prepare 422kg of high tryptophan fermented feed dry base.

Example 5

S2, preparation of tryptophan concentrate:

S3, preparation of fermentation source tryptophan concentrated solution:

S4, preparing the high tryptophan fermented feed:

118kg of tryptophan concentrated solution, 197kg of wet substrate of glycolysis residue meal and 100kg of commercial tryptophan are mixed and then dried or respectively dried and then mixed to prepare 270kg of high tryptophan fermented feed dry substrate.

Example 6

S2, preparation of tryptophan concentrate:

S3, preparation of fermentation source tryptophan concentrated solution:

S4, preparing the high tryptophan fermented feed:

and mixing 175kg of tryptophan concentrated solution, 546kg of wet substrate of glycolysis residue meal and 200kg of commercial tryptophan, and then drying or respectively drying and then mixing to prepare 622kg of high-tryptophan fermented feed dry substrate.

The contents of main active substances in the high tryptophan fermented feeds prepared in the first, example 1, example 2, example 3 and example 5 are shown in the following table 1.

Table 1, example 2, example 3 and example 5 the high tryptophan fermented feed products prepared had the nutrient content

Secondly, the effect of feeding weaned pigs by the high tryptophan fermented feed prepared in the example 1.

To investigate the feeding effect of the high tryptophan fermented feed prepared in example 1 on weaned piglets, 500 growing up ternary hybrid healthy piglets of 23 days old were selected and randomly divided into 5 groups according to the principle that the body weight and the gestation ratio are similar and the sex is half, each group has 10 repetitions, and each repetition has 10 pigs. The control group did not add high tryptophan fermented feed. The test groups respectively add high tryptophan fermentation feed according to 10 kg/ton, 20 kg/ton and 30 kg/ton to prepare weaned piglet feed. The protein and energy of each group of feed are similar. Piglets were weaned for 23 days with a trial period of 30 days. The test results are as follows:

TABLE 2 influence of the high tryptophan fermented feed prepared in example 1 on the productivity of weaned piglets

Item	Control group	10 kg/ton group	20 kg/ton group	30 kg/ton group
					Daily gain (g)	386.78	425.6	434.52	438.69
Daily food intake (g)	564.70	608.61	617.02	618.55
					Meat ratio of materials	1.46	1.43	1.42	1.41

As can be seen from Table 2, after 10 kg/ton, 20 kg/ton and 30 kg/ton of high tryptophan fermented feed is added into the daily ration, the daily gain of the weaned piglets is respectively increased by 10.04%, 12.34% and 13.42% compared with the control group, the feed-meat ratio is respectively reduced by 2.10%, 2.82% and 3.55% compared with the control group, and the feed intake is respectively increased by 7.77%, 9.26% and 9.53% compared with the control group. The results show that: the high tryptophan fermented feed can obviously improve the production performance of weaned piglets.

And thirdly, the effect of feeding the high tryptophan fermented feed prepared in the example 2 on growing pigs.

To investigate the feeding effect of the high tryptophan fermented feed prepared in example 2 on growing pigs, 84 heads (59 and 60-day-old piglets) of Du-Dai three-way hybrid piglets with similar body weights (body weights of 18.9 + -0.85 kg) were selected and randomly divided into a test group and a control group. The test group had 42 pigs (sow 22 and boar 20), and the control group had 42 pigs (sow 19 and boar 23). The piglets in the two groups were similar in weight (significance analysis was not significant), and each group was provided with 7 replicates, each replicate 6.

The piglet basal diet nutrition water average of the test group and the piglet basal diet of the control group are the same and are powder, the control group does not contain high tryptophan fermented feed, and the test group is added with 20 kg/ton of high tryptophan fermented feed.

Test pre-test period: the test starts from 2018, 9, 16 and 10 days, works such as prevention, inoculation, insect expelling and the like are well done, and the test is started after the end of the pre-test.

Test pre-correction test period: the test formally started on 7.10.2018 and formally ended on 30.11.2018 for 54 days.

After 20 kg/ton of high tryptophan fermented feed is added into daily ration, the average end weight of 42 piglets in a test group is 38.06kg, the average end weight of a control group is 35.71kg, the number of the piglets in the test group is 2.35kg more than that in the control group, and the average end weight is improved by 6.58% (P is less than 0.05); the average weight of the test group is increased by 19.24kg, the average weight of the control group is increased by 16.66kg, the number of the test group is increased by 2.58kg compared with the control group, and the increase is 15.49 percent (P is less than 0.01); the average daily gain of the test group is 356.30g, the average daily gain of the control group is 308.52g, the test group has 47.78g more than the control group, and the increase is 15.49 percent (P is less than 0.01). See table 3 for details.

TABLE 3 Effect of the high tryptophan fermented feed prepared in example 2 on the daily gain of growing pigs

Group of

Number of heads

Days of experiment

Initial weight/head/kg

End weight/head kg

Total weight gain/head/kg

Daily gain/head/g

Control group

42

54

19.05

35.71±1.69

16.66±1.04

308.52±64

Test group

42

54

18.82

38.06±2.85^*

19.24±1.67^**

356.30±111^**

Note:^**indicates significance of intra-column differences (P)<0.01)，^*Indicates significance of intra-column differences (P)<0.05)。

As can be seen from Table 3, after 20 kg/ton of high tryptophan fermented feed is added into the daily ration, the feed intake of each test group is 45.21kg, the feed intake of each control group is 40.32kg, the feed intake of the test group is 40.89kg. more than that of the control group, and the feed intake is increased by 12.13% (P is less than 0.05); the feed conversion rate of the test group is 2.35, which is 0.07 percent less than that of the control group 2.42, and the feed conversion rate of the test group is 2.89 percent less than that of the control group (P is more than 0.05). See table 4 for details.

TABLE 4 influence of the high tryptophan fermented feed prepared in example 2 on the feed conversion ratio of growing pigs

Group of	Weight gain/head/kg	Feed intake/head/kg	Feed conversion ratio	Cost of feed/yuan	Cost/yuan/1 kg weight gain
						Control group	16.66	40.32±1.23	2.42±0.49	60.48	3.63
Test group	19.24	45.21±2.35*	2.35±0.83	67.82	3.52

Note: calculated according to the price of the local feed raw materials at that time, the feed is 1.508 yuan/kg.

Fourthly, the effect of feeding broiler chickens with the high tryptophan fermentation feed prepared in example 3.

To investigate the feeding effect of the high tryptophan fermented feed prepared in example 6 on broilers, 800 AA broilers of 1 day old were selected and randomly divided into 4 groups of 10 replicates each, and 20 replicates each, based on the principle of half weight and sex. The broiler feed is prepared by adding 5 kg/ton, 10 kg/ton and 15 kg/ton of the feed, and the high tryptophan fermentation feed is not added in the ration of the control group. The protein and energy of each group of feed are similar. The test period was 42 days. The test results are shown in table 5:

TABLE 5 influence of the high tryptophan fermented feed prepared in example 3 on the productivity of broiler chickens

As can be seen from Table 5, after 5 kg/ton, 10 kg/ton and 15 kg/ton of the high tryptophan fermented feed are added to the broiler feed, the daily gain of the broiler is respectively increased by 8.54%, 11.44% and 12.58% (P <0.01) compared with the control group, the feed intake is respectively increased by 6.52%, 8.48% and 11.52% (P <0.05 or P <0.01) compared with the control group, and the feed-meat ratio is respectively reduced by 4.08%, 5.10% and 4.59(P <0.05) compared with the control group. The results show that: the high tryptophan fermented feed can obviously improve the production performance of the broiler chicken.

Fifthly, the effect of feeding the laying hens by the high tryptophan fermented feed prepared in the example 5.

To investigate the feeding effect of the high tryptophan fermented feed prepared in example 10 on laying hens, 800 roman laying hens at the egg laying peak period of 34 weeks were selected and randomly divided into 4 groups of 10 replicates each and 20 replicates each. Adding the feed at 5 kg/ton, 10 kg/ton and 15 kg/ton to prepare the laying hen feed, wherein the high tryptophan fermented feed is not added in the daily ration of the control group. The protein and energy of each group of feed are similar. The test period was 28 days. The test results are shown in table 6:

as can be seen from Table 6, the laying rates of the Roman layer chickens fed with 5 kg/ton, 10 kg/ton and 15 kg/ton of the high tryptophan fermented feeds were respectively increased by 3.69%, 6.92% and 7.72% compared with the control group (P >0.05 or P <0.05) and the laying rates of the Roman layer chickens fed with 38 weeks were respectively increased by 4.50%, 8.97% and 9.35% compared with the control group (P >0.05 or P < 0.05). The results show that: the high tryptophan fermented feed can obviously improve the egg laying performance of the laying hens.

TABLE 6 influence of the high tryptophan fermented feed prepared in example 5 on egg laying performance of Roman hens

The foregoing is illustrative and explanatory only and is not intended to be exhaustive or to limit the invention to the precise embodiments described, and various modifications, additions, and substitutions may be made by those skilled in the art without departing from the spirit of the invention or exceeding the scope defined by the claims.

Claims

1. A preparation method of high tryptophan fermented feed is characterized by comprising the following steps:

s2, preparation of fermentation source tryptophan concentrated solution:

s3, preparation of fermentation source tryptophan concentrated solution:

s4, preparing the high tryptophan fermented feed:

mixing the fermentation source tryptophan concentrated solution and the fermentation residue meal wet base, directly producing the high tryptophan fermentation feed wet base, or mixing and drying, or respectively drying and mixing to prepare the high tryptophan fermentation feed dry base.

2. The method for preparing a high tryptophan fermented feed according to claim 1, wherein the tryptophan fermentation broth and the wet tryptophan thallus base are prepared by the following steps in step S1: preparing tryptophan fermentation liquor and tryptophan thallus wet base or crushing starch-containing products, adding water for size mixing, adding amylase for enzymolysis, liquefying and saccharifying, and performing solid-liquid separation to obtain sugar residue wet base and residue-free saccharified liquid; adding nutrient solution required by microorganisms into the residue-free saccharified solution, sterilizing at high temperature, cooling to a suitable temperature required by the growth of the microorganisms, inoculating a tryptophan-producing strain, introducing sterile air, fermenting for several hours, and directly preparing tryptophan fermentation liquor.

3. The method for preparing a fermented feed with high tryptophan content according to claim 1, wherein in the step S3, the fermented dreg pulp wet base is prepared by the following method: mixing one or more of the sugar residue wet base and the tryptophan thallus wet base with feed raw materials, clean water, composite probiotic bacteria liquid and composite enzyme, and performing enzymolysis and fermentation to obtain the glycolysis residue meal wet base.

4. The method of claim 1, wherein the fermented feed with high tryptophan content is prepared by the following steps in step S4: mixing the fermentation source tryptophan concentrated solution, the fermentation residue meal wet base and commercial feed non-fermentation source tryptophan, directly producing the high tryptophan fermentation feed wet base, or mixing and drying the wet base or respectively drying and mixing the dry bases to prepare the high tryptophan fermentation feed dry base.

5. The method for preparing a high tryptophan fermented feed according to claim 1, wherein the nutrient solution required by the microorganism is one or more of yeast extract, yeast powder, peptone, beef extract, corn steep liquor, ammonium sulfate, magnesium sulfate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium sulfate, vitamin B1 and biotin;

the feed raw materials are one or more of bean pulp, cotton pulp, rapeseed pulp, peanut pulp, fish meal, blood meal, plasma protein, blood cell protein, milk powder, lactose, whey powder, wheat gluten, corn protein powder, rice protein powder, DDGS, soybeans, peas, red beans, corns, rice, broken rice, wheat, bran, rice bran, flour, oats, barley, sweet potatoes, soybean protein isolate, soybean protein concentrate, pasture meal, Chinese herbal medicines, fruits and vegetables, sugar molasses syrup and byproducts of the food processing industry, or puffed products of the feed raw materials;

the probiotic bacteria liquid is one or more of bacillus coagulans, bacillus subtilis, bacillus licheniformis, saccharomyces cerevisiae, brewer's yeast, aroma-producing yeast, lactobacillus acidophilus, bifidobacterium, enterococcus faecium, pediococcus acidilactici, lactobacillus plantarum and lactobacillus delbrueckii;

the complex enzyme is one or more of high-temperature alpha amylase, saccharifying enzyme, fungal enzyme, beta amylase, mannase, xylanase, glucanase, pectinase and protease.

6. The method for preparing a high tryptophan fermented feed according to claim 1, wherein the fermentation source tryptophan concentrate is prepared under the following conditions in step S2:

7. The method for preparing a high tryptophan fermented feed according to claim 1, wherein the fermentation source tryptophan concentrate is prepared under the following conditions in step S2:

8. The method for preparing the high tryptophan fermented feed according to claim 1, wherein the conditions for preparing the wet fermented dreg pulp in the step S3 are as follows:

9. The method for preparing the high tryptophan fermented feed according to claim 1, wherein the conditions for preparing the wet fermented dreg pulp in the step S3 are as follows:

10. The method of claim 1, wherein the conditions for preparing the high tryptophan fermented feed in the step S4 are as follows: