A peptide for promoting rice bran fermentation for goose feed and its application

Abstract

The purpose of the present invention is to provide a peptide for promoting rice bran fermentation for goose feed and its application. Fresh rice bran is enzymatically hydrolyzed and fermented to obtain an enzymatic fermentation complex, which is used for goose feeding. The produced small peptides and beneficial intestinal bacteria can provide better nutrition, are easy to absorb, improve disease resistance, avoid oxidation of rice bran, and extend the storage period of rice bran.

Description

Technical Field

[0001] The invention relates to the technical field of biological peptides and goose feed, and in particular to a peptide for promoting rice bran fermentation for goose feed and application thereof. Background Art

[0002] Alternative feeds and agricultural byproducts have become an important solution to the shortage of feed in the livestock industry. Rice bran has become a very attractive option for poultry feed due to its relatively high nutritional content. However, several factors limit the use of rice bran in poultry nutrition, such as its low protein and high fiber and fat content and the presence of anti-nutrients such as phytic acid. Fermentation products can improve the quality of rice bran and thus improve the performance of rice bran.

[0003] Although enzymatic fermentation is a common treatment method, current research is relatively confusing, with unclear effects or only basic research without practical application. For example, in the study "Effects of adding enzyme preparations to rice bran diets on metabolic hormones and biochemical indices in goslings", Ai Xiaojie et al. showed that enzyme preparations increased the blood insulin level of goslings by 40.82% (P<0.01), increased the blood glucose level by 4.76% (P>0.05), decreased the glucagon level by 31.18% (P<0.01), and increased the ratio of insulin to glucagon 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 with the control group. ); (3) Plasma T3 level increased by 24.42% (P<0.05) and thyroid stimulating hormone level decreased by 8.85% (P>0.05) compared with the control. Enzyme preparations can affect the body's metabolic hormone levels and change the metabolism of sugar and protein by eliminating the anti-nutritional effects of arabinoxylan in feed, thereby promoting the growth of goslings. At the same time, Ai Xiaojie et al. also proved the effect of adding enzyme preparations to rice bran diet on the viscosity and pH of goslings' small intestinal chyme, the effect of adding enzyme preparations to rice bran diet on the development of goslings' digestive organs, and the effect of adding enzyme preparations to rice bran diet on the activity of pancreatic and chyme enzymes in goslings. CN200910086085 extracts protein from rice bran or directly uses it as protein feed. The resulting mixture is too rough at the protein level and also contains rice bran oil; CN201410069513 often requires rice bran and other nutrients to be fermented together before feeding, which increases costs.

[0004] Therefore, the present invention focuses on analyzing the polypeptides in rice bran, and analyzes the functions of multiple small peptides through modern biological methods, develops them in a targeted and purposeful manner to reduce blindness, and conducts animal experiments on the peptides with the highest scores to achieve the effect of precise feeding. Summary of the invention

[0005] The invention aims to provide a peptide for promoting the fermentation of rice bran for goose feed, which is easy to absorb, improves the disease resistance rate, avoids the oxidation of rice bran, and prolongs the storage period of rice bran.

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

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

[0008] The first step of enzymatic hydrolysis is to use at least one of flavor protease, amylase, saccharifying enzyme and cellulase for enzymatic hydrolysis.

[0009] The first step of enzymatic hydrolysis is to use at least two of flavor protease, amylase, saccharifying enzyme and cellulase for enzymatic hydrolysis.

[0010] The second enzymatic hydrolysis step uses at least one of a neutral protease and an alkaline protease for enzymatic hydrolysis.

[0011] The second step of enzymolysis is performed using neutral protease and alkaline protease.

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

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

[0014] The invention obtains small peptides with low molecular weight and high yield through two enzymatic hydrolysis of rice bran, and also obtains rich oil and polysaccharide compounds for later research and development. In addition, the invention focuses on analyzing the polypeptides in the rice bran, and analyzes the functions of a plurality of small peptides obtained by modern biological means, so as to develop them in a targeted and purposeful manner and reduce blindness. The peptides with the highest scores are subjected to animal experiments to achieve the effect of accurate feeding. The generated small peptides and corresponding intestinal beneficial bacteria can provide better nutrition for geese, are easy to absorb, and improve the disease resistance rate. The invention increases body weight and increases the average weight gain while keeping the fluctuation of the average feed consumption small and having a high feed conversion rate. The IMPc, the average muscle protein content, the leg muscle rate and the chest muscle rate are all better than those of the control group, the average intramuscular fat is reduced, the weight can be increased evenly, the meat has high nutritional value, high protein content, low fat content, good fresh taste and flavor, and good egg quality, and is suitable for the healthy diet advocated by 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, the eggshell will not be too thin to break, and the eggshell is not too thick to dissipate heat, there is no dead embryo egg, the egg yolk has good luster, the protein content is high, the egg yolk ratio is moderate and has not increased significantly, and it can also be suitable for patients with high cholesterol; the rice bran can also be processed in time to avoid oxidation of the rice bran and extend the storage period of the rice bran. DETAILED DESCRIPTION

[0015] The embodiments of the present invention are further described below in conjunction with the examples.

[0016] Example 1

[0017] Fresh rice bran is washed, sterilized with bacteria and enzymes at high temperature, filtered, water is added (material-water ratio is 10:1), stirred evenly, enzymes are added in sequence for enzymolysis (substance to be enzymolyzed: enzyme 50:2), and the enzymolysis is divided into two steps. The temperature of the enzymolysis process is optimized according to the temperature control device of the actual factory. In the first step, flavor protease, amylase, saccharifying enzyme, and cellulase (1.5:1:1:1) are used for enzymolysis, pH is adjusted to 3-8, fully stirred for 0.5-3 hours, filtered, water is added, and stirred. In the second step, neutral protease and alkaline protease are used for enzymolysis, pH is adjusted to 8-10, fully stirred for 0.5-3 hours, and allowed to stand for 2-5 hours. The enzymatic residues were removed by centrifuge filtration, filtered and purified, and then filtered through a membrane to retain the small molecule peptide complex liquid below 10,000 Daltons, vacuum freeze-dried, and sent for detection and analysis. The key proteins (immunoglobulin, myosin, actin, MAPK, TGF-β) were analyzed by bioinformatics, computer models, molecular docking methods, and nutritional protein scores to obtain the highest-scoring MSSGDSQLAK, which was used for the subsequent nutritional evaluation of peptide addition to goose feed and the antibacterial evaluation of peptides.

[0018] Example 2

[0019] Nutritional evaluation of peptides and antimicrobial evaluation of peptides

[0020] 90 healthy 10-day-old geese were selected from the farm, with similar weights (within 10 g), and randomly divided into three groups: a control group (ordinary commercial feed (containing corn, wheat bran, soybean meal, vegetable oil, green feed, crude protein, calcium, phosphorus, and complex vitamins)), an experimental group 1, a small molecule peptide complex liquid group (the above ordinary commercial feed + 2.5% small molecule peptide complex liquid group), and an experimental group 2, a preferred peptide group (2.5% MSSGDSQLAK + the above ordinary commercial feed), 30 geese in each group, with free food and water, body weight and feed intake recorded weekly, interval values ​​displayed as minimum and maximum values, and special cases are marked separately.

[0021] After 70 days of feeding, 10 animals were randomly selected from each group, and feces were taken for bacterial species analysis and parasite detection. The animals were fasted for 6 hours before slaughter, weighed before slaughter, and inosinic acid, muscle and intramuscular fat were analyzed. 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] After being raised for 70 days, the experimental groups 1 and 2 had good feeding uniformity, good feather gloss, bright color, good appetite, and strong disease resistance. There was no Riemerella disease, goose plague, influenza, goose paratyphoid, diarrhea, conjunctivitis, emaciation, fowl cholera, aspergillosis, or tapeworm disease in goose and duck plague. Three goose in the control group had diarrhea (high numbers of Escherichia coli and Salmonella were found).

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

[0024]

[0025]

[0026] The determination of inosinic acid (related to meat flavor) (IMP) content in goose muscle is carried out in accordance with the National Standard of the People's Republic of China (GB / T19676-2005). Since the determination value of inosinic acid is greatly affected by temperature and time, the corrected inosinic acid is used for statistical analysis. That is, the corrected inosinic acid (IMPc) content is equal to the sum of the determination 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 intramuscular fat (IMF) content in goose muscle was carried out in accordance with the National Standard of the People's Republic of China (GB / T14772-2008). The IMF content in this experiment refers to the percentage of fat in muscle dry matter, and a large amount of unsaturated fatty acids C16:0C18:0 were found. Generally, 10 weeks is the time when unsaturated fatty acids are the highest during the growth period. Other test items are commonly used test methods and will not be described one by one.

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

[0029]

[0030]

[0031] For the rest, the average age at first laying, daily egg production rate, average egg weight, egg quality, average fertilization rate, average hatching rate, average brooding survival rate, and average egg-laying hens survival rate were calculated for the reared chicks until the egg-laying period.

[0032] Two goose eggs were randomly selected from each goose. Egg weight, egg shape index, eggshell strength, eggshell thickness, egg specific gravity, yolk color, albumen height, Haugh unit, yolk ratio and egg white ratio were measured according to the "Terminology and Statistical Methods for Poultry Production Performance" (NY / T823-2004). Multivariate analysis was performed and scored according to the 100-point system, with each item being 10 points (such as egg weight of experimental group 1 / egg weight of experimental group 2*10, and then each item was added up, and the control group and experimental group 1 were evaluated with experimental group 2 as 100 points to obtain the total score of relative egg quality. Other indicators can be calculated normally.

[0033] Table 3 Egg production in each group

[0034]

[0035]

[0036] Throughout the whole process, experimental groups 1 and 2 had good uniformity of feeding, bright color, strong appetite, strong disease resistance, low feed consumption, low cost, uniform weight gain (about 105g per week), and no Riemerella disease, goose plague, influenza, goose paratyphoid, diarrhea, conjunctivitis, emaciation, fowl cholera, aspergillosis, and tapeworm disease. The growth intensity was the highest at about 1 week of age, and the weight gain was faster at 3 to 10 weeks of age. The vigorous period of shin length and shin circumference growth and development was 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 highest at about 1-3 weeks of age, and the weight gain was faster at 4 to 10 weeks of age. The vigorous period of shin length and shin circumference growth and development was 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, it would not be too thin to break the egg, it would not be too thick to dissipate heat, there was no dead embryo egg, the yolk had good luster, the protein content was high, the yolk ratio was moderate and did not increase significantly, and it was also suitable for patients with high cholesterol.

[0037] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A peptide for goose feed, characterized in that: The sequence is MSSGDSQLAK.

2. A peptide for goose feed according to claim 1, characterized in that: The peptides are obtained by enzymatic hydrolysis of rice bran.

Images