A method for preparing a peptide fermented for a goose feed using rice bran

Abstract

The present application aims to provide a preparation method of rice bran fermented peptide for goose feed, which comprises the following steps: fermenting fresh rice bran to obtain an enzymatic fermentation compound, and feeding the goose with the enzymatic fermentation compound, so that the small peptides and intestinal beneficial bacteria produced can provide better nutrition, are easy to absorb, improve the disease resistance rate, and can avoid oxidation of the rice bran and prolong the storage period of the rice bran.

Description

Technical Field

[0001] This invention relates to the field of bioactive peptides and goose feed technology, specifically to a method for preparing peptides for goose feed from fermented rice bran and their application. Background Technology

[0002] Alternative feeds and agricultural byproducts have become important solutions to feed shortages in livestock farming. Rice bran, due to its relatively high nutritional content, is a very strong option for poultry feed. However, several factors limit the utilization of rice bran in poultry nutrition, such as its low protein and high fiber and fat content, as well as the presence of anti-nutritional agents such as phytic acid. Fermentation products can improve the quality of rice bran, thereby improving its performance.

[0003] Although enzymatic fermentation is a common treatment method, current research is relatively chaotic, with unclear effects or only basic research without practical application. For example, in the study of "the effect of adding enzyme preparations to rice bran diets on metabolic hormones and biochemical indicators in goslings", Ai Xiaojie et al. showed that enzyme preparations increased blood insulin levels in goslings by 40.82% (P<0.01), blood glucose levels by 4.76% (P>0.05), decreased glucagon levels by 31.18% (P<0.01), and increased the insulin to glucagon ratio by 104.18% (P<0.01); (2) plasma alanine aminotransferase activity increased by 38.39% (P<0.05) and uric acid concentration decreased by 10.60% (P<0.05) compared to the control. (3) Plasma T3 level increased by 24.42% (P<0.05) and thyroid-stimulating hormone level decreased by 8.85% (P>0.05). Enzyme preparations can affect the body's metabolic hormone levels and change the metabolism of sugar and protein by relieving the anti-nutritional effects of arabinoxylan and other substances in feed, thereby promoting the growth of goslings. At the same time, Ai Xiaojie et al. also proved the effects of adding enzyme preparations to rice bran diet on the viscosity and pH of gosling small intestine chyme, the effects of adding enzyme preparations to rice bran diet on the development of gosling digestive organs, and the effects of adding enzyme preparations to rice bran diet on the pancreas and chyme enzyme activity of goslings. CN200910086085 Extracting protein from rice bran or using it directly as protein feed results in a mixture that is too coarse at the protein level and also contains rice bran oil; CN201410069513 Rice bran often needs to be fermented with other nutrients before feeding, which increases costs.

[0004] Therefore, this invention focuses on analyzing the polypeptides in rice bran. By analyzing the functions of multiple small peptides using modern biological methods, targeted development can be carried out to reduce blind spots. The peptide with the highest score is then used in animal experiments to achieve precise feeding results. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing peptides from fermented rice bran for goose feed, which is easy to absorb, improves disease resistance, avoids oxidation of rice bran, and extends the shelf life of rice bran.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] The present invention provides a method for preparing peptides for goose feed from fermented rice bran, wherein the peptide sequence is MSSGDSQLAK, and the peptides are obtained by enzymatic hydrolysis of rice bran in two steps.

[0008] The first step of the enzymatic hydrolysis is carried out using at least one of flavor protease, amylase, saccharifying enzyme, and cellulase.

[0009] The first step of the enzymatic hydrolysis is performed using at least two of the following: flavor protease, amylase, saccharifying enzyme, and cellulase.

[0010] The second step of the enzymatic hydrolysis is performed using at least one of neutral protease and alkaline protease.

[0011] The second step of the enzymatic hydrolysis is performed using neutral protease and alkaline protease.

[0012] Another aspect of the present invention provides the application of peptides derived from rice bran enzymatic hydrolysis and fermentation for use in goose feed. The application also includes feeding poultry, including chickens, ducks, and rabbits, and specifically geese.

[0013] The beneficial effects of this invention are:

[0014] This invention utilizes two enzymatic hydrolysis processes in rice bran to obtain low-molecular-weight peptides with high yield. It also yields abundant oils and polysaccharides for future research and development. Furthermore, it focuses on analyzing the polypeptides in rice bran, employing modern biological methods to analyze the functions of multiple small peptides, enabling targeted development and reducing randomness. The peptides with the highest scores are then used in animal experiments to achieve precise feeding. The resulting small peptides and corresponding beneficial intestinal bacteria provide geese with better nutrition, easier absorption, and improved disease resistance. Weight gain and average weight gain are increased while maintaining minimal fluctuations in average feed consumption and high feed conversion rate. IMPc, average muscle protein content, leg muscle percentage, and pectoral muscle percentage are all superior to the control group, with reduced average intramuscular fat, resulting in uniform weight gain, high-nutritional-value meat with high protein and low fat content, excellent flavor, and good egg quality, making it suitable for the healthy diet advocated in modern society. It has high feed utilization, low cost, high egg production rate, good egg quality, high eggshell strength, moderate eggshell thickness (not too thin and the egg will break, not too thick and it will not be difficult to dissipate heat), no dead embryos, good yolk color, high protein content, and a moderate yolk ratio without significant increase. It can also be used for patients with high cholesterol. It can also process rice bran in a timely manner to avoid oxidation of rice bran and extend the storage period of rice bran. Detailed Implementation

[0015] The embodiments of the present invention will be further described below with reference to examples.

[0016] Example 1

[0017] Wash fresh rice bran, sterilize it at high temperature to remove impurities and enzymes, filter it, add water (material-to-water ratio 10:1), stir well, and then add enzymes sequentially for enzymatic hydrolysis (hydrolysate: enzyme 50:2). The enzymatic hydrolysis is divided into two steps. The temperature of the enzymatic hydrolysis process is optimized according to the actual temperature control device of the factory. In the first step, enzymatic hydrolysis is carried out with flavor protease, amylase, saccharifying enzyme, and cellulase (1.5:1:1:1), and the pH is adjusted to 3-8. Stir thoroughly for 0.5-3 hours, filter, add water, and stir. In the second step, enzymatic hydrolysis is carried out with neutral protease and alkaline protease, and the pH is adjusted to 8-10. Stir thoroughly for 0.5-3 hours and let stand for 2-5 hours. Centrifugation and filtration were used to remove enzymatic residues. The purified solution was then filtered through a membrane to retain small molecule peptide complexes with a molecular weight of less than 10,000 Daltons. The solution was then freeze-dried under vacuum and sent for analysis. Through bioinformatics analysis of key proteins (immunoglobulins, myosin, actin, MAPK, TGF-β), computer models and molecular docking methods were used to analyze and score the nutritional proteins. The highest-scoring MSSGDSQLAK was obtained and used for subsequent evaluation of peptide nutrition and antibacterial properties in goose feed.

[0018] Example 2

[0019] Nutritional evaluation and antimicrobial evaluation of peptides

[0020] Ninety healthy 10-day-old goslings with similar weights (less than 10g) were selected from the farm and randomly divided into three groups: a control group (common commercial feed containing corn, wheat bran, soybean meal, vegetable oil, green feed, crude protein, calcium, phosphorus, and compound vitamins), experimental group 1 (small molecule peptide compound solution group, the above-mentioned common commercial feed + 2.5% small molecule peptide compound solution group), and experimental group 2 (preferred peptide group, 2.5% MSSGDSQLAK + the above-mentioned common commercial feed), with 30 goslings in each group. They had free access to food and water. Their weight and feed intake were measured and recorded weekly. The interval values ​​are displayed as the minimum and maximum values. Special cases are marked separately.

[0021] Animals were raised for 70 days. Ten animals were randomly selected from each group, and fecal samples were collected for bacterial count analysis and parasite detection. Animals were fasted for 6 hours before slaughter and weighed before slaughter. Inosinic acid, muscle mass, and intramuscular fat were analyzed. Weight (kg), average feed intake (g), average weight gain (g), IMPc, average muscle protein content, leg muscle percentage, pectoral muscle percentage, and average intramuscular fat were calculated.

[0022] After 70 days of feeding, experimental groups 1 and 2 showed good feeding uniformity, good feather luster and bright color, strong appetite, and strong disease resistance. No goose or duck plague, goose plague, influenza, goose paratyphoid, diarrhea, conjunctivitis, emaciation, fowl cholera, aspergillosis, or tapeworm disease were observed. In contrast, three birds in the control group developed diarrhea (with high levels of Escherichia coli and Salmonella found).

[0023] Table 1. Analysis of fecal bacterial species and parasite detection in each group.

[0024]

[0025]

[0026] The determination of inosinic acid (IMP), which is related to the umami flavor of goose muscle, was carried out in accordance with the People's Republic of China National Standard (GB / T19676-2005). Since the measured values ​​of inosinic acid are significantly affected by temperature and time, a corrected inosinic acid method was used for statistical analysis. That is, the corrected inosinic acid (IMPc) content is equal to the sum of the measured values ​​of ADP, AMP, IMP, INO, and HYP divided by their respective molecular weights, multiplied by the molecular weight of IMP.

[0027] The intramuscular fat (IMF) content in goose muscle was determined according to the People's Republic of China National Standard (GB / T14772-2008). The IMF content in this experiment represents the percentage of fat in the muscle dry matter. A large amount of unsaturated fatty acids (C16:0 C18:0) was found, and the highest levels of unsaturated fatty acids were generally observed around week 10 of the growth period. Other detection methods were commonly used and will not be detailed further.

[0028] Table 2 shows the weight, inosine monophosphate (IMPc), muscle and intramuscular fat content of each group.

[0029]

[0030]

[0031] For the remaining breeding to the egg-laying period, calculate the average age at first laying, daily egg production rate, average egg weight, egg quality, average fertilization rate, average hatching rate, average brooding period survival rate, and average egg-laying period hen survival rate.

[0032] Two goose eggs were randomly selected from each goose's eggs. Egg weight, egg shape index, eggshell strength, eggshell thickness, egg specific gravity, yolk color, albumen height, Haugh units, yolk ratio, and egg white ratio were measured according to the "Nomenclature and Measurement Statistical Methods for Poultry Production Performance" (NY / T823-2004). Multifactor analysis was performed, and each item was scored out of 100 points (e.g., egg weight of experimental group 1 / egg weight of experimental group 2 * 10, and then the scores of each item were added together, with experimental group 2 as 100 points to compare and evaluate the control group and experimental group 1, and obtain the relative total egg quality score. Other indicators were calculated normally.

[0033] Table 3 Egg production data for each group

[0034]

[0035]

[0036] Throughout the entire process, experimental groups 1 and 2 showed good feeding uniformity, bright egg color, strong appetite, strong disease resistance, low feed consumption, low cost, and uniform weight gain (approximately 105g per week). No goose or duck plague, goose plague, influenza, goose paratyphoid, diarrhea, conjunctivitis, emaciation, fowl cholera, aspergillosis, or tapeworm disease were observed. Growth intensity peaked around 1 week of age, with rapid weight gain from 3 to 10 weeks. The peak growth period for shank length and shank circumference occurred around 5 weeks of age, earlier than the control group. The control group showed peak growth intensity from 1 to 3 weeks of age, with rapid weight gain from 4 to 10 weeks. The peak growth period for shank length and shank circumference occurred around 7 weeks of age. Egg production was high, egg quality was good, eggshell strength was high, and eggshell thickness was moderate—not too thin and prone to breakage, nor too thick and difficult to dissipate heat. No dead embryos were found. Egg yolk color was good, egg white content was high, and the yolk-to-egg ratio was moderate without significant increase. This method is also suitable for patients with high cholesterol.

[0037] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of a rice bran fermentation peptide in the preparation of goose feed, characterized in that, The peptide sequence is MSSGDSQLAK, and the application is for feeding geese.

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