Preparation method of peptide fermented by rice bran and used for goose feed

Abstract

According to the preparation method of the rice bran fermented peptide for the goose feed, fresh rice bran is subjected to enzymolysis and fermentation to obtain an enzymolysis and fermentation compound which is used for feeding geese, the generated small peptide and intestinal beneficial bacteria can provide better nutrition, are easy to absorb and improve the disease resistance rate, and can also avoid oxidation of the rice bran; the storage period of the rice bran is prolonged.

Description

Technical Field

[0001] The present invention relates to the technical fields of biological peptides and goose feed, and particularly relates to a preparation method and application of peptides for goose feed fermented from rice bran. Background Art

[0002] Alternative feeds and agricultural by-products have become important solutions to the shortage of livestock feed. Rice bran has become a very powerful choice for poultry feed due to its relatively high nutrient content. However, several factors limit the utilization of rice bran in poultry nutrition, such as its low protein, high fiber and fat content, and the presence of anti-nutritional agents such as phytic acid. Fermentation products can improve the quality of rice bran and thus improve its performance.

[0003] Although enzymatic fermentation is a common treatment method, current research is relatively chaotic, the effects are unclear or only basic research has been carried out without practical application. For example, "Effects of adding enzyme preparations to rice bran diets on metabolic hormones and biochemical indices of goslings", Ai Xiaojie et al. showed that the enzyme preparations increased the blood insulin level of goslings by 40.82% (P<0.01), the blood glucose level by 4.76% (P>0.05), the glucagon level decreased by 31.18% (P<0.01), and the ratio of insulin to glucagon increased by 104.18% (P<0.01); (2) the activity of plasma alanine aminotransferase increased by 38.39% compared with the control (P<0.05), and the uric acid concentration decreased by 10.60% (P<0.05); (3) the plasma T3 level increased by 24.42% compared with the control (P<0.05), and the thyroid stimulating hormone level decreased by 8.85% (P>0.05). The enzyme preparations can affect the levels of metabolic hormones in the body and change the metabolism of sugar and protein by removing the anti-nutritional effects of arabinoxylan and other substances in the feed, thus promoting the growth of goslings. At the same time, Ai Xiaojie et al. also proved the effects of adding enzyme preparations to rice bran diets on the viscosity and pH of gosling intestinal chyme, the development of gosling digestive organs, and the activities of gosling pancreas and chyme enzymes. CN200910086085 extracts proteins from rice bran or directly uses it as a protein feed, and the obtained mixture is too rough at the protein level and also contains rice bran oil; CN201410069513 often requires co-fermentation of rice bran and other nutrients before feeding, resulting in increased costs.

[0004] Therefore, the present invention focuses on analyzing the polypeptides in rice bran, analyzing the functions of multiple small peptides obtained by modern biological means, developing them purposefully to reduce blindness, and conducting animal experiments on the peptides with the highest scores to achieve the effect of precise feeding. Summary of the Invention

[0005] The object of the present invention is to provide a preparation method of peptides for goose feed fermented from rice bran, which is easy to absorb, can improve the disease resistance rate, and can also avoid the oxidation of rice bran and extend the storage period of rice bran.

[0006] To achieve the above object, the present invention is realized through the following technical solutions:

[0007] On the one hand, the present invention provides a preparation method of peptides for goose feed fermented from rice bran. The peptide sequence is MSSGDSQLAK, and the peptide is obtained by enzymatic hydrolysis of rice bran. The enzymatic hydrolysis is divided into two steps.

[0008] In the first step of the enzymatic hydrolysis, at least one of flavor protease, amylase, glucoamylase, and cellulase is used for enzymatic hydrolysis.

[0009] In the first step of the enzymatic hydrolysis, at least two of flavor protease, amylase, glucoamylase, and cellulase are used for enzymatic hydrolysis.

[0010] In the second step of the enzymatic hydrolysis, at least one of neutral protease and alkaline protease is used for enzymatic hydrolysis.

[0011] In the second step of the enzymatic hydrolysis, neutral protease and alkaline protease are used for enzymatic hydrolysis.

[0012] On the other hand, the present invention provides an application of peptides for goose feed after enzymatic hydrolysis and fermentation of rice bran. The application also includes the feeding of poultry, and the poultry includes the feeding of chickens, ducks, and rabbits, and the poultry includes the feeding of geese.

[0013] The beneficial effects of the present invention are:

[0014] Through the two-step enzymatic hydrolysis of rice bran, small peptides with low molecular weight are obtained, with a high yield. Abundant oils and polysaccharide compounds are also obtained for later R & D and utilization. Moreover, the polypeptides in rice bran are analyzed emphatically. The functions of multiple small peptides obtained through modern biological means are analyzed, and targeted development is carried out to reduce blindness. The peptide with the highest score is used in animal experiments to achieve the effect of precise feeding. The small peptides produced and the corresponding beneficial intestinal bacteria can provide better nutrition for geese, are easy to absorb, and improve the disease resistance rate; while increasing body weight and the average weight gain, the fluctuation of the average feed consumption is small, and the feed conversion rate is high; IMPc, the average muscle protein content, the leg muscle rate, and the breast muscle rate are all better than those of the control group, and the average intramuscular fat decreases, enabling uniform weight gain, high meat nutritional value, high protein content, low fat content, good umami flavor, good egg quality, and being suitable for the healthy diet advocated in modern society. The feed utilization rate is high, the cost is low, the egg production rate is high, the egg quality is good, the eggshell strength is high, the eggshell thickness is moderate, not too thin to break eggs and not too thick to dissipate heat easily, there are no dead embryo eggs, the egg yolk color degree is good, the protein content is high, the egg yolk ratio is moderate without significant increase, and it is also applicable to patients with high cholesterol; it is also possible to process rice bran in a timely manner to avoid the oxidation of rice bran and extend the storage period of rice bran. Detailed implementation mode

[0015] The following further illustrates the implementation mode of the present invention in conjunction with the embodiments.

[0016] Example 1

[0017] Wash the fresh rice bran, filter it after high-temperature sterilization of miscellaneous bacteria and enzymes, add water (material-water ratio 10:1), stir evenly, and add enzymes in sequence for enzymatic hydrolysis (hydrolyzate: enzyme 50:2). The enzymatic hydrolysis is divided into 2 steps. The temperature is optimized and selected according to the temperature control device of the actual factory during the enzymatic hydrolysis process. In the first step, use flavor protease, amylase, glucoamylase, and cellulase (1.5:1:1:1) for enzymatic hydrolysis, adjust the pH = 3 - 8, stir well for 0.5 - 3 hours, filter, add water and stir. In the second step, use neutral protease and alkaline protease for enzymatic hydrolysis, adjust the pH = 8 - 10, stir well for 0.5 - 3 hours, and let it stand for 2 - 5 hours. Centrifuge and filter to remove the enzymatic hydrolysis residue, filter and purify, and then filter through a filter membrane to intercept the small molecule peptide complex solution with a molecular weight below 10,000 Daltons, and perform vacuum freeze-drying, send it out for detection and analysis, and analyze the key proteins (immunoglobulin, myosin, actin, MAPK, TGF-β) computer model and molecular docking method through bioinformatics analysis and obtain the MSSGDSQLAK with the highest score for subsequent evaluation of the peptide nutrition added to goose feed and the antibacterial evaluation of the peptide.

[0018] Example 2

[0019] Peptide nutrition evaluation and antibacterial evaluation of peptides

[0020] 90 healthy meat geese at 10 days old with little difference in weight (within 10 g) were selected from a farm and randomly divided into three groups: a control group (fed with common commercially available feed (containing corn, wheat bran, soybean meal, vegetable oil, green feed, crude protein, calcium, phosphorus, and compound vitamins)), an experimental group 1 (small molecule peptide complex solution group, i.e., the above common commercially available feed + 2.5% small molecule peptide complex solution), and an experimental group 2 (preferred peptide group, 2.5% MSSGDSQLAK + the above common commercially available feed). There were 30 geese in each group. They had free access to food and water. Their body weights were measured weekly and feed intakes were recorded. The interval values were shown as the lowest and highest values, and special cases were marked separately.

[0021] When they were raised to 70 days old, 10 geese were randomly selected from each group, and their feces were taken for analysis of the number of bacteria and parasite detection. They were fasted 6 hours before slaughter, weighed before slaughter, and inosine monophosphate, muscle, and intramuscular fat were analyzed. The weight (kg), average feed consumption (g), average weight gain (g), IMPc, average muscle protein content, leg muscle rate, breast muscle rate, and average intramuscular fat were calculated.

[0022] When raised to 70 days old, the geese in experimental groups 1 and 2 had good feeding uniformity, with shiny feathers, bright colors, strong appetites, and strong disease resistance. There were no cases of Riemerella anatipestifer, goose plague, influenza, goose paratyphoid, diarrhea, conjunctivitis, emaciation, fowl cholera, aspergillosis, or cestodiasis. In the control group, 3 cases of diarrhea occurred (relatively high numbers of Escherichia coli and Salmonella were found).

[0023] Table 1 Analysis of the number of bacteria in feces and parasite detection for each group

[0024]

[0025]

[0026] The determination of the inosine monophosphate (related to meat umami) (IMP) content in goose muscle was carried out according to the national standard of the People's Republic of China (GB / T19676 - 2005). Since the measured value of inosine monophosphate is greatly affected by temperature and time, corrected inosine monophosphate was used for statistical analysis. That is, the content of corrected inosine monophosphate (IMPc) is equal to the sum of the measured values of ADP, AMP, IMP, INO, and HYP divided by their respective molecular weights and then multiplied by the molecular weight of IMP.

[0027] The determination of the intramuscular fat (IMF) content in goose muscle was carried out according to the national standard of the People's Republic of China (GB / T14772 - 2008). The IMF content in this experiment was the percentage of fat in muscle dry matter, and a large amount of unsaturated fatty acids C16:0 and C18:0 were found. Generally, 10 weeks is the time when the unsaturated fatty acids are the highest during the growth period. The other detection items used common detection methods and will not be elaborated one by one.

[0028] Table 2 Weights, inosinic acid IMPc, muscle and intramuscular fat contents of each group.

[0029]

[0030]

[0031] The rest were raised until the laying period to calculate the average age at first lay, daily egg production rate, average egg weight, egg quality, average fertilization rate, average hatching rate, average survival rate during the brooding period, and average survival rate of female poultry during the laying period.

[0032] Randomly select 2 goose eggs from the eggs of each goose, and measure the egg weight, egg shape index, eggshell strength, eggshell thickness, egg specific gravity, yolk color, albumen height, Haugh unit, yolk ratio and egg white ratio in accordance with "Nomenclature and Measurement Statistics Methods for Poultry Production Performance" (NY / T 823-2004), conduct multivariate analysis, and score according to a 100-point system, with 10 points for each item (such as experimental group 1 egg weight / experimental group 2 egg weight * 10, and then sum each item. Taking experimental group 2 as 100 points as the control, evaluate the control group and experimental group 1 to obtain the total relative egg quality score, and the other indicators can be calculated normally.

[0033] Table 3 Laying performance of each group

[0034]

[0035]

[0036] Throughout the whole process, the feeding uniformity of experimental groups 1 and 2 was good, with fresh color, strong appetite, strong disease resistance, less feed consumption, low cost, and uniform weight gain (about 105 g per week). There were no occurrences of Riemerella anatipestifer, goose plague, influenza, goose paratyphoid, diarrhea, conjunctivitis, emaciation, fowl cholera, aspergillosis, and cestodiasis. The growth intensity was the largest at about 1 week of age, the weight increased relatively fast from 3 to 10 weeks of age, and the peak periods of the growth and development of shank length and shank circumference were at about 5 weeks of age. The development period was earlier than that of the control group. The growth intensity of the control group was the largest at about 1 - 3 weeks of age, the weight increased relatively fast from 4 to 10 weeks of age, and the peak periods of the growth and development of shank length and shank circumference were at about 7 weeks of age. The egg production rate was high, the egg quality was good, the eggshell strength was high, the eggshell thickness was moderate, not too thin to break or too thick to dissipate heat, there were no dead embryo eggs, the yolk color was good, the protein content was high, and the yolk ratio was moderate without significant increase. It was also applicable to patients with high cholesterol.

[0037] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A preparation method of peptides for goose feed fermented from rice bran, characterized in that, The sequence of the peptide is MSSGDSQLAK.

2. The preparation method of a peptide for goose feed fermented from rice bran according to claim 1, characterized in that, The peptide is obtained by enzymatic hydrolysis of rice bran.

3. The preparation method of a peptide for goose feed fermented from rice bran according to claim 2, characterized in that, The enzymatic hydrolysis is divided into two steps.

4. The preparation method of a peptide for goose feed fermented from rice bran according to claim 3, characterized in that, In the first step of the enzymatic hydrolysis, at least one of flavor protease, amylase, glucoamylase, and cellulase is used for enzymatic hydrolysis.

5. The preparation method of a peptide for goose feed fermented from rice bran according to claim 3, characterized in that, In the first step of the enzymatic hydrolysis, at least two of flavor protease, amylase, glucoamylase, and cellulase are used for enzymatic hydrolysis.

6. The preparation method of a peptide for goose feed fermented from rice bran according to claim 3, characterized in that, In the second step of the enzymatic hydrolysis, at least one of neutral protease and alkaline protease is used for enzymatic hydrolysis.

7. The preparation method of a peptide for goose feed fermented from rice bran according to claim 3, characterized in that, In the second step of the enzymatic hydrolysis, neutral protease and alkaline protease are used for enzymatic hydrolysis.

8. Use of the peptide prepared by the preparation method of the peptide for goose feed fermented from rice bran according to claims 1-7, characterized in that, The application also includes application in the feeding of poultry.

9. Use of the peptide prepared by the preparation method of the peptide for goose feed fermented from rice bran according to claim 8, characterized in that, The poultry includes application in the feeding of chickens, ducks, and rabbits.

10. Use of the peptide prepared by the preparation method of the peptide for goose feed fermented from rice bran according to claim 8, characterized in that, The poultry includes application in the feeding of geese.

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