CN118716462A - Preparation method of animal feed with fermented litchi pomace and probiotics - Google Patents
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Abstract
Description
技术领域Technical Field
本发明属于生物技术和饲料加工技术领域,具体涉及一种发酵荔枝果渣协同益生菌的动物饲料的制备方法。The invention belongs to the technical field of biotechnology and feed processing, and particularly relates to a method for preparing animal feed prepared by fermenting litchi pomace and coordinating probiotics.
背景技术Background Art
荔枝果渣是荔枝加工过程中的副产品,如果直接丢弃或不当处理,不仅会造成资源浪费,还可能对环境造成污染。荔枝果渣中含有丰富的纤维素、半纤维素、矿物质、维生素以及多种活性物质使得其可以做为动物饲料利用,其含有的大量酚类化合物和蛋白质成分,为动物提供丰富养分的同时,还具有一定的抗氧化作用,预防动物疾病的发生,使得动物能够健康成长。Litchi pomace is a byproduct of litchi processing. If it is directly discarded or improperly handled, it will not only cause a waste of resources, but also may pollute the environment. Litchi pomace is rich in cellulose, hemicellulose, minerals, vitamins and a variety of active substances, which makes it possible to use it as animal feed. It contains a large amount of phenolic compounds and protein components, which not only provide rich nutrients for animals, but also have a certain antioxidant effect, prevent the occurrence of animal diseases, and enable animals to grow healthily.
然而,由于果渣含水量高、易霉变,且其中成分结构复杂,大分子蛋白含量高,多酚也多为不溶性酚,直接用于饲料动物不仅难以充分消化吸收,还会影响动物的消化吸收和适口性。However, due to the high water content and easy moldiness of the pomace, the complex structure of its ingredients, the high content of large molecular protein, and the fact that most polyphenols are insoluble phenols, it is not only difficult to fully digest and absorb if used directly as feed for animals, but it will also affect the animals' digestion, absorption and palatability.
因此,若想将荔枝果渣用于动物饲料,需先对其进行预处理,提高可溶性多酚含量及蛋白质利用率,从而提高动物饲料的营养价值,使得动物能够健康成长。Therefore, if litchi pomace is to be used as animal feed, it must be pretreated first to increase the soluble polyphenol content and protein utilization rate, thereby improving the nutritional value of animal feed and allowing animals to grow healthily.
发明内容Summary of the invention
本发明的目的是提供一种发酵荔枝果渣协同益生菌的动物饲料的制备方法,提高了动物饲料中可吸收营养物质的含量和有益酶活性,同时提高了适口性,促进了动物生长发育。The purpose of the present invention is to provide a method for preparing animal feed of fermented litchi pomace and probiotics, which increases the content of absorbable nutrients and beneficial enzyme activity in the animal feed, while improving the palatability and promoting the growth and development of animals.
为了实现上述目的,本发明提供了以下技术方案:In order to achieve the above object, the present invention provides the following technical solutions:
所述发酵荔枝果渣协同益生菌的动物饲料的制备方法,包括以下步骤:The method for preparing the animal feed of fermented litchi pomace and probiotics comprises the following steps:
S1、将荔枝果渣粉碎后加入水调节含水量为60%~70%,得到荔枝果渣液,调节其pH为5.5~6.5后,添加碳酸钙,搅拌均匀后在80~90℃下均质10~20min后,得到预处理荔枝果渣;S1, after crushing litchi pomace, adding water to adjust the water content to 60% to 70%, obtaining litchi pomace liquid, adjusting the pH value to 5.5 to 6.5, adding calcium carbonate, stirring evenly, and homogenizing at 80 to 90° C. for 10 to 20 minutes to obtain pretreated litchi pomace;
S2、将预处理荔枝果渣、益生菌和复合酶混匀,在30~40℃下发酵48~72h,即得发酵后的荔枝果渣;S2, mixing the pretreated litchi pomace, probiotics and complex enzyme, and fermenting at 30-40° C. for 48-72 h to obtain fermented litchi pomace;
S3、将发酵后的荔枝果渣和饲料混合均匀,再添加解脂假丝酵母和益生元,在25~30℃下二次发酵24~48h,得到动物饲料。S3, mixing the fermented litchi pomace and feed evenly, adding Candida lipolytica and prebiotics, and performing secondary fermentation at 25-30° C. for 24-48 hours to obtain animal feed.
优选的,所述碳酸钙和荔枝果渣液的固液比为5~7g:1L。Preferably, the solid-liquid ratio of the calcium carbonate to the litchi pomace liquid is 5-7 g:1 L.
发明人发现,将荔枝果渣调节在一个合适的pH范围内,同时加入适量碳酸钙,能够提高最终产物中营养物质含量。这可能是因为本发明中特定的益生菌和复合酶在特定的pH范围内生长最为旺盛,但是在发酵期间,益生菌对葡萄糖和果糖的过程中会产生大量的乳酸,导致荔枝果渣的pH值逐渐下降,影响益生菌尤其是干酪乳杆菌的生长。发明人意外发现,通过添加适量的碳酸钙,能够中和产生的乳酸,减缓pH下降,从而优化发酵环境,提高乳酸菌的活性和发酵效率,促进乳酸菌的生长,进而提高荔枝果渣的发酵效果,提高发酵后的荔枝果渣中营养物质含量。The inventor found that by adjusting the litchi pomace within a suitable pH range, adding an appropriate amount of calcium carbonate, the nutrient content in the final product can be increased. This may be because specific probiotics and complex enzymes grow the most vigorously within a specific pH range in the present invention, but during fermentation, a large amount of lactic acid will be produced in the process of probiotics to glucose and fructose, causing the pH value of the litchi pomace to gradually decline, affecting the growth of probiotics, especially Lactobacillus casei. The inventor unexpectedly found that by adding an appropriate amount of calcium carbonate, the lactic acid produced can be neutralized, the pH decline is slowed down, thereby optimizing the fermentation environment, improving the activity and fermentation efficiency of lactic acid bacteria, promoting the growth of lactic acid bacteria, and then improving the fermentation effect of litchi pomace, improving the nutrient content in the litchi pomace after fermentation.
优选的,所述益生菌为植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的混合物。Preferably, the probiotics are a mixture of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae.
优选的,所述植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的质量比为(1~3):(2~4):(3~5):(4~6):(1~3):1;进一步优选的,为2:3:4:5:2:1。Preferably, the mass ratio of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae is (1-3):(2-4):(3-5):(4-6):(1-3):1; more preferably, it is 2:3:4:5:2:1.
优选的,所述植物乳杆菌中活性乳酸菌含量≥80亿CFU/克。Preferably, the content of active lactic acid bacteria in the Lactobacillus plantarum is ≥ 8 billion CFU/g.
优选的,所述乳酸片球菌中有效活菌数含量≥80亿CFU/克。Preferably, the effective viable bacteria count in the Pediococcus acidilactici is ≥ 8 billion CFU/g.
优选的,所述枯草芽孢杆菌中活性乳酸菌含量≥900亿CFU/克。Preferably, the content of active lactic acid bacteria in the Bacillus subtilis is ≥ 90 billion CFU/g.
优选的,所述干酪乳杆菌中活性乳酸菌含量≥80亿CFU/克。Preferably, the content of active lactic acid bacteria in the Lactobacillus casei is ≥ 8 billion CFU/g.
在一些优选的方案中,所述植物乳杆菌、枯草芽孢杆菌和干酪乳杆菌均购自西安西海生物科技有限公司。In some preferred embodiments, the Lactobacillus plantarum, Bacillus subtilis and Lactobacillus casei are all purchased from Xi'an Xihai Biotechnology Co., Ltd.
优选的,所述地衣芽孢杆菌中活性乳酸菌含量≥900亿CFU/克。Preferably, the content of active lactic acid bacteria in the Bacillus licheniformis is ≥ 90 billion CFU/g.
在一些优选的方案中,所述乳酸片球菌、地衣芽孢杆菌均购自山东中科嘉亿生物工程有限公司。In some preferred embodiments, the Pediococcus acidilactici and Bacillus licheniformis are purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.
优选的,所述酿酒酵母的酵母活性细胞数≥150亿/g。Preferably, the number of active yeast cells of the brewer's yeast is ≥ 15 billion/g.
在一些优选的方案中,所述酿酒酵母购自山东英菲达生物科技有限公司。In some preferred embodiments, the brewer's yeast is purchased from Shandong Yingfeida Biotechnology Co., Ltd.
发明人发现,选用特定的益生菌,且所述植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的质量比为(1~3):(2~4):(3~5):(4~6):(1~3):1时,能够显著增加发酵后的荔枝果渣中多酚,尤其是自由酚和可溶性酚含量的同时,提高蛋白酶的活性,进而促进营养物质的消化吸收。这可能是因为植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌协同作用,一方面在发酵过程中能够利用荔枝果渣中的糖类等营养物质进行生长繁殖,并通过其代谢活动产生各种酶类,这些酶类能够分解荔枝果渣中的复杂成分,促进荔枝果渣中结合酚的释放,增加可溶性酚的含量。另一方面益生菌的发酵作用还可能改变荔枝果渣中膳食纤维等成分的结构,使其与多酚类物质之间的结合键断裂,从而增加可溶性酚含量。枯草芽孢杆菌、地衣芽孢杆菌、酿酒酵母复配,枯草芽孢杆菌、地衣芽孢杆菌代谢互补,它们各自产生的蛋白酶具有不同的底物特异性和作用机制,共同作用于荔枝果渣中的蛋白质,从而实现对蛋白质更全面的分解和利用,同时,酿酒酵母具有环境调控作用,其在发酵过程中产生的代谢产物有助于激活某些蛋白酶的活性。丰富的活性高的蛋白酶有利于将果渣中大分子蛋白物质分解成易吸收的小分子物质,使得发酵后的荔枝果渣更适宜添加到动物饲料中。The inventors found that when a specific probiotic is selected and the mass ratio of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis, and Saccharomyces cerevisiae is (1-3): (2-4): (3-5): (4-6): (1-3): 1, the polyphenols in the fermented litchi pomace, especially the free phenol and soluble phenol content, can be significantly increased, while the activity of protease is improved, thereby promoting the digestion and absorption of nutrients. This may be because Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, and Bacillus licheniformis work synergistically, on the one hand, can utilize nutrients such as sugars in litchi pomace to grow and reproduce during the fermentation process, and produce various enzymes through their metabolic activities, which can decompose the complex components in litchi pomace, promote the release of bound phenols in litchi pomace, and increase the content of soluble phenols. On the other hand, the fermentation of probiotics may also change the structure of components such as dietary fiber in litchi pomace, break the bond between it and polyphenols, thereby increasing the content of soluble phenols. Bacillus subtilis, Bacillus licheniformis and Saccharomyces cerevisiae are compounded, and the metabolism of Bacillus subtilis and Bacillus licheniformis is complementary. The proteases produced by them have different substrate specificity and action mechanisms, and they work together to act on the protein in the litchi pomace, thereby achieving a more comprehensive decomposition and utilization of the protein. At the same time, Saccharomyces cerevisiae has an environmental regulation effect, and the metabolites produced by it during the fermentation process help to activate the activity of certain proteases. The rich and highly active proteases are conducive to decomposing the macromolecular protein substances in the pomace into easily absorbable small molecules, making the fermented litchi pomace more suitable for adding to animal feed.
优选的,所述复合酶为纤维素酶和果胶酶。Preferably, the complex enzyme is cellulase and pectinase.
优选的,所述纤维素酶和果胶酶的质量比为(4~6):1;进一步优选的,为5:1。Preferably, the mass ratio of the cellulase to the pectinase is (4-6):1; more preferably, it is 5:1.
优选的,所述纤维素酶的酶活≥10000u/g。Preferably, the enzymatic activity of the cellulase is ≥10000u/g.
在一些优选的方案中,所述纤维素酶购自宁夏夏盛实业集团有限公司。In some preferred embodiments, the cellulase is purchased from Ningxia Xiasheng Industrial Group Co., Ltd.
优选的,所述果胶酶的酶活≥25000u/g。Preferably, the enzymatic activity of the pectinase is ≥25000u/g.
在一些优选的方案中,所述果胶酶购自宁夏夏盛实业集团有限公司。In some preferred embodiments, the pectinase is purchased from Ningxia Xiasheng Industrial Group Co., Ltd.
发明人发现,选用特定的纤维素酶和果胶酶作为复合酶,且纤维素酶和果胶酶的质量比为(4~6):1时,能够在保证发酵后的荔枝果渣营养成分的同时,改善果渣的质地和口感,使其更易于被动物消化吸收。这可能是因为纤维素酶和果胶酶在荔枝果渣发酵过程中可以共同作用于细胞壁,增强对细胞壁成分的降解效果。纤维素酶主要降解纤维素,而果胶酶则主要降解果胶,两者协同作用可以使细胞壁更加彻底地降解,从而释放更多的营养物质,如葡萄糖等还原糖,进而提高了发酵后的荔枝果渣的营养价值及口感。The inventors found that by selecting specific cellulase and pectinase as composite enzymes, and the mass ratio of cellulase to pectinase is (4-6):1, the texture and taste of the fermented litchi pomace can be improved while ensuring the nutritional content of the fermented litchi pomace, making it easier for animals to digest and absorb. This may be because cellulase and pectinase can act together on the cell wall during the fermentation of litchi pomace, enhancing the degradation effect of cell wall components. Cellulase mainly degrades cellulose, while pectinase mainly degrades pectin. The synergistic effect of the two can make the cell wall degrade more thoroughly, thereby releasing more nutrients, such as reducing sugars such as glucose, thereby improving the nutritional value and taste of the fermented litchi pomace.
优选的,所述益生菌和复合酶的质量比为1:(2~4);进一步优选的,为1:3。Preferably, the mass ratio of the probiotics to the complex enzyme is 1:(2-4); further preferably, it is 1:3.
发明人发现,当益生菌和复合酶的质量比为1:(2~4)时,由于纤维素酶和果胶酶的协同作用,细胞壁的降解更为迅速和彻底,这为复合菌微生物提供了更多的营养物质,使得发酵过程可以更加高效地进行,缩短发酵时间并提高发酵后的荔枝果渣的营养成分,促使动物健康成长。The inventors found that when the mass ratio of probiotics to complex enzymes is 1:(2-4), due to the synergistic effect of cellulase and pectinase, the degradation of cell walls is more rapid and thorough, which provides more nutrients for the complex bacterial microorganisms, allowing the fermentation process to proceed more efficiently, shortening the fermentation time and increasing the nutritional content of the fermented lychee pomace, thereby promoting the healthy growth of animals.
优选的,所述益生菌和复合酶的添加量为预处理荔枝果渣质量的1%~5%;进一步优选的,为3%。Preferably, the added amount of the probiotics and complex enzyme is 1% to 5% of the mass of the pretreated litchi pomace; more preferably, it is 3%.
所述饲料不做具体限制,可为动物食用即可,例如玉米、麦麸、豆粕等。The feed is not particularly limited and can be anything that animals can eat, such as corn, wheat bran, soybean meal, etc.
优选的,所述发酵后的荔枝果渣的添加量为饲料质量的100%~120%。Preferably, the added amount of the fermented litchi pomace is 100% to 120% of the feed mass.
在一些优选的方案中,所述解脂假丝酵母购自北京万佳标准物质研发中心。In some preferred embodiments, the Candida lipolytica is purchased from Beijing Wanjia Standard Material Research and Development Center.
所述益生元为甘露寡糖和果寡糖。The prebiotics are manno-oligosaccharides and fructo-oligosaccharides.
所述甘露寡糖和果寡糖的质量比为(1~3):1;进一步优选的,为2:1。The mass ratio of manno-oligosaccharide to fructo-oligosaccharide is (1-3):1; more preferably, it is 2:1.
所述甘露寡糖中有效成分含量≥92%,蛋白质含量≤1%,菌落总数≤1000CFU/g。The mannooligosaccharide has an effective component content of ≥92%, a protein content of ≤1%, and a total colony count of ≤1000 CFU/g.
所述果寡糖中有效成分含量≥95%,菌落总数<10CFU/g。The content of effective ingredients in the fructooligosaccharide is ≥95%, and the total number of colonies is <10 CFU/g.
在一些优选的方案中,所述甘露寡糖和果寡糖均购自河南冠琪生物科技有限公司。In some preferred embodiments, the manno-oligosaccharides and fructo-oligosaccharides are purchased from Henan Guanqi Biotechnology Co., Ltd.
所述解脂假丝酵母菌和益生元的质量比为(2~4):1;进一步优选的,为3:1。The mass ratio of the Candida lipolytica to the prebiotics is (2-4):1; more preferably, it is 3:1.
所述解脂假丝酵母菌的添加量为发酵后的荔枝果渣和饲料总质量的0.5%~1.5%。The addition amount of the Candida lipolytica is 0.5% to 1.5% of the total mass of the fermented litchi pomace and feed.
在一些优选的方案中,选用质量比为(2~4):1的解脂假丝酵母和益生元对饲料和发酵后的荔枝果渣进行二次发酵,能够进一步提高蛋白酶活性,促进动物的生长发育。这可能是因为经过一次发酵后,荔枝果渣中仍残留部分植酸、纤维素、单宁等抗营养物质影响果渣的适口性和营养价值,同时,豆粕、麦麸、玉米等传统饲料中含有抗营养因子、纤维素等成分,不易被动物吸收。发明人发现,通过添加一定量的解脂假丝酵母,不仅能够降解荔枝果渣和豆粕中的抗营养物质,还能产生多种酶类,和残留的复合酶共同降解荔枝果渣、玉米和麦麸等饲料中的纤维素、果胶等大分子物质,将其转化为小分子物质的同时,产生的蛋白酶还能够提高蛋白酶活性和蛋白质利用率,从而改善饲料的适口性和动物的肠道健康,提高饲料的营养价值。In some preferred schemes, the feed and the fermented litchi pomace are subjected to secondary fermentation by using Candida lipolytica and prebiotics in a mass ratio of (2-4):1, which can further improve the activity of protease and promote the growth and development of animals. This may be because after the first fermentation, some anti-nutritional substances such as phytic acid, cellulose, and tannins still remain in the litchi pomace, affecting the palatability and nutritional value of the pomace. At the same time, traditional feeds such as soybean meal, wheat bran, and corn contain anti-nutritional factors, cellulose and other ingredients that are not easily absorbed by animals. The inventors found that by adding a certain amount of Candida lipolytica, not only can the anti-nutritional substances in litchi pomace and soybean meal be degraded, but also a variety of enzymes can be produced, which can degrade macromolecules such as cellulose and pectin in litchi pomace, corn, and wheat bran together with the residual composite enzyme, and convert them into small molecules. At the same time, the produced protease can also improve the protease activity and protein utilization, thereby improving the palatability of the feed and the intestinal health of the animal, and improving the nutritional value of the feed.
此外,选用适量的甘露寡糖和果寡糖作为益生元,两种益生元不仅能够促进益生菌的增殖和生长,与解脂假丝酵母之间还有协同作用,一方面果寡糖为解脂假丝酵母提供生长所需的营养物质,促进其在肠道内的定植和繁殖,另一方面甘露寡糖与解脂假丝酵母通过竞争肠道内的营养和定植位点,共同抑制病原菌的生长,从而进一步增强肠道健康,提高动物肠道对营养物质的吸收,从而促进动物的生长发育。In addition, appropriate amounts of manno-oligosaccharides and fructo-oligosaccharides are selected as prebiotics. The two prebiotics can not only promote the proliferation and growth of probiotics, but also have a synergistic effect with Candida lipolytica. On the one hand, fructo-oligosaccharides provide Candida lipolytica with the nutrients needed for growth and promote its colonization and reproduction in the intestine. On the other hand, manno-oligosaccharides and Candida lipolytica jointly inhibit the growth of pathogens by competing for nutrients and colonization sites in the intestine, thereby further enhancing intestinal health and improving the absorption of nutrients by the animal intestine, thereby promoting the growth and development of animals.
本发明还包括所述发酵荔枝果渣协同益生菌的动物饲料的制备方法制备得到的动物饲料。The present invention also includes animal feed prepared by the method for preparing animal feed of fermented litchi pomace and probiotics.
与现有技术相比,本发明的优点和有益效果为:Compared with the prior art, the advantages and beneficial effects of the present invention are:
1.本发明提供一种发酵荔枝果渣协同益生菌的动物饲料的制备方法,对荔枝果渣进行发酵,提高了发酵荔枝果渣中可溶性酚等营养成分,同时提高了荔枝果渣中蛋白酶的活性,从而有利于将果渣中大分子蛋白物质分解成易吸收的小分子物质,将该发酵荔枝果渣应用于制备动物饲料,并进行二次发酵,能够进一步降解抗营养物质和纤维素等成分,提高易吸收的营养物质含量,为动物提供充足的营养,促进动物生长发育。此外,本发明提高了荔枝果渣的综合利用率,增加荔枝产业经济附加值。1. The present invention provides a method for preparing animal feed of fermented litchi pomace and synergistic probiotics, wherein the litchi pomace is fermented, the soluble phenol and other nutrients in the fermented litchi pomace are increased, and the activity of protease in the litchi pomace is increased, so as to facilitate the decomposition of macromolecular protein substances in the pomace into easily absorbable small molecules, and the fermented litchi pomace is applied to prepare animal feed, and secondary fermentation is performed, which can further degrade anti-nutrient substances and cellulose and other components, increase the content of easily absorbable nutrients, provide sufficient nutrition for animals, and promote animal growth and development. In addition, the present invention improves the comprehensive utilization rate of litchi pomace and increases the economic added value of the litchi industry.
2.本发明将荔枝果渣调节在一个合适的pH范围内,同时加入适量碳酸钙,能够提高最终产物中营养物质含量。2. The present invention adjusts the litchi pomace to a suitable pH range and adds a proper amount of calcium carbonate, thereby increasing the nutrient content in the final product.
3.本发明选用特定的益生菌,且所述植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的质量比为(1~3):(2~4):(3~5):(4~6):(1~3):1时,能够显著增加发酵后的荔枝果渣中多酚,尤其是自由酚和可溶性酚含量的同时,提高蛋白酶的活性,进而促进营养物质的消化吸收。3. The present invention selects specific probiotics, and when the mass ratio of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae is (1-3):(2-4):(3-5):(4-6):(1-3):1, the polyphenols in the fermented litchi pomace, especially the contents of free phenol and soluble phenol, can be significantly increased, while the activity of protease is improved, thereby promoting the digestion and absorption of nutrients.
4.本发明选用特定的纤维素酶和果胶酶作为复合酶,且纤维素酶和果胶酶的质量比为(4~6):1时,能够在保证发酵后的荔枝果渣营养成分的同时,改善果渣的质地和口感,使其更易于被动物消化吸收。4. The present invention selects specific cellulase and pectinase as the composite enzyme, and when the mass ratio of cellulase to pectinase is (4-6):1, the texture and taste of the fermented litchi pomace can be improved while ensuring the nutritional components of the fermented litchi pomace, making it easier for animals to digest and absorb.
5.本发明选用质量比为1:(2~4)的复合菌和复合酶,缩短发酵时间并提高发酵后的荔枝果渣的营养成分,促使动物健康成长。5. The present invention selects composite bacteria and composite enzymes in a mass ratio of 1: (2-4), shortens the fermentation time and improves the nutritional content of the fermented litchi pomace, thereby promoting the healthy growth of animals.
6.本发明选用质量比为(2~4):1的解脂假丝酵母和益生元对饲料和发酵后的荔枝果渣进行二次发酵,能够进一步提高蛋白酶活性,促进动物的生长发育。6. The present invention uses Candida lipolytica and prebiotics in a mass ratio of (2-4):1 to perform secondary fermentation on feed and fermented litchi pomace, which can further improve protease activity and promote the growth and development of animals.
7.本发明选用适量的甘露寡糖和果寡糖作为益生元,两种益生元不仅能够促进益生菌的增殖和生长,与解脂假丝酵母还有协同作用,提高动物肠道对营养物质的吸收,从而促进动物的生长发育。7. The present invention selects appropriate amounts of manno-oligosaccharides and fructo-oligosaccharides as prebiotics. The two prebiotics can not only promote the proliferation and growth of probiotics, but also have a synergistic effect with Candida lipolytica to improve the absorption of nutrients by the animal intestine, thereby promoting the growth and development of the animal.
具体实施方式DETAILED DESCRIPTION
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其它实施例,都属于本发明保护的范围。The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
本发明使用的原料均为市售,具体的:The raw materials used in the present invention are all commercially available, specifically:
植物乳杆菌中活性乳酸菌含量约100亿CFU/克,枯草芽孢杆菌中活性乳酸菌含量约1000亿CFU/克,干酪乳杆菌中活性乳酸菌含量约100亿CFU/克,均购自西安西海生物科技有限公司。The active lactic acid bacteria content in Lactobacillus plantarum is about 10 billion CFU/g, the active lactic acid bacteria content in Bacillus subtilis is about 100 billion CFU/g, and the active lactic acid bacteria content in Lactobacillus casei is about 10 billion CFU/g, all of which are purchased from Xi'an Xihai Biotechnology Co., Ltd.
乳酸片球菌中有效活菌数含量约100亿CFU/克,地衣芽孢杆菌中活性乳酸菌含量约1000亿CFU/克,均购自山东中科嘉亿生物工程有限公司。The effective live bacteria count of Pediococcus acidilactici is about 10 billion CFU/g, and the active lactic acid bacteria count of Bacillus licheniformis is about 100 billion CFU/g, both of which were purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.
酿酒酵母的酵母活性细胞数约200亿/g,购自山东英菲达生物科技有限公司。The number of active yeast cells of Saccharomyces cerevisiae was about 20 billion/g, which was purchased from Shandong Infida Biotechnology Co., Ltd.
纤维素酶的酶活约11000u/g,果胶酶的酶活约30000u/g,均购自宁夏夏盛实业集团有限公司。The enzymatic activity of cellulase is about 11000u/g, and the enzymatic activity of pectinase is about 30000u/g, both of which were purchased from Ningxia Xiasheng Industrial Group Co., Ltd.
解脂假丝酵母购自北京万佳标准物质研发中心。Candida lipolytica was purchased from Beijing Wanjia Standard Material Research and Development Center.
甘露寡糖中有效成分含量≥92%,蛋白质含量≤1%,菌落总数≤1000CFU/g;果寡糖中有效成分含量≥95%,菌落总数<10CFU/g,均购自河南冠琪生物科技有限公司。Manno-oligosaccharide has an effective ingredient content of ≥92%, a protein content of ≤1%, and a total colony count of ≤1000 CFU/g; fructooligosaccharide has an effective ingredient content of ≥95%, and a total colony count of <10 CFU/g, all of which were purchased from Henan Guanqi Biotechnology Co., Ltd.
实施例1Example 1
本实施例提供了一种发酵荔枝果渣协同益生菌的动物饲料的制备方法,步骤为:This embodiment provides a method for preparing animal feed with fermented litchi pomace and probiotics, the steps are as follows:
S1、将荔枝果渣粉碎后加入水调节含水量为65%,得到荔枝果渣液,调节其pH为6后,添加碳酸钙,搅拌均匀后在85℃下均质15min后,得到预处理荔枝果渣;S1, crushing litchi pomace and adding water to adjust the water content to 65% to obtain litchi pomace liquid, adjusting the pH value to 6, adding calcium carbonate, stirring evenly and homogenizing at 85° C. for 15 min to obtain pretreated litchi pomace;
S2、将预处理荔枝果渣、益生菌和复合酶混匀,在35℃下发酵60h,即得发酵后的荔枝果渣;S2, mixing the pretreated litchi pomace, probiotics and complex enzyme, and fermenting at 35° C. for 60 h to obtain fermented litchi pomace;
S3、将发酵后的荔枝果渣和饲料混合均匀,再添加解脂假丝酵母和益生元,在28℃下二次发酵36h,得到动物饲料。S3. Evenly mix the fermented litchi pomace and feed, add Candida lipolytica and prebiotics, and perform secondary fermentation at 28° C. for 36 hours to obtain animal feed.
所述碳酸钙和荔枝果渣液的固液比为6g:1L。The solid-to-liquid ratio of the calcium carbonate to the litchi pomace liquid is 6g:1L.
所述益生菌为植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的混合物。The probiotics are a mixture of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae.
所述植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的质量比为2:3:4:5:2:1。The mass ratio of the Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae is 2:3:4:5:2:1.
所述复合酶为纤维素酶和果胶酶。The complex enzyme is cellulase and pectinase.
所述纤维素酶和果胶酶的质量比为5:1。The mass ratio of the cellulase to the pectinase is 5:1.
所述益生菌和复合酶的质量比为1:3。The mass ratio of the probiotics to the complex enzyme is 1:3.
所述益生菌和复合酶的添加量为预处理荔枝果渣质量的3%。The added amount of the probiotics and the complex enzyme is 3% of the mass of the pretreated litchi pomace.
所述饲料为玉米、麦麸、豆粕。The feed is corn, wheat bran and soybean meal.
所述玉米、麦麸、豆粕的质量比为2:2:1。The mass ratio of the corn, wheat bran and soybean meal is 2:2:1.
所述发酵后的荔枝果渣的添加量为饲料质量的110%。The added amount of the fermented litchi pomace is 110% of the mass of the feed.
所述益生元为甘露寡糖和果寡糖。The prebiotics are manno-oligosaccharides and fructo-oligosaccharides.
所述甘露寡糖和果寡糖的质量比为2:1。The mass ratio of manno-oligosaccharide to fructo-oligosaccharide is 2:1.
所述解脂假丝酵母菌和益生元的质量比为3:1。The mass ratio of the Candida lipolytica to the prebiotics is 3:1.
所述解脂假丝酵母菌的添加量为发酵后的荔枝果渣和饲料总质量的1%。The addition amount of the Candida lipolytica is 1% of the total mass of the fermented litchi pomace and feed.
实施例2Example 2
本实施例与实施例1的区别为:所述碳酸钙和荔枝果渣液的固液比为7g:1L。The difference between this embodiment and embodiment 1 is that the solid-liquid ratio of the calcium carbonate to the litchi pomace liquid is 7 g:1 L.
实施例3Example 3
本实施例与实施例1的区别为:所述植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的质量比为3:4:4:4:1:1。The difference between this embodiment and embodiment 1 is that the mass ratio of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae is 3:4:4:4:1:1.
对比例1Comparative Example 1
本对比例与实施例1的区别为:未调节荔枝果渣pH值。The difference between this comparative example and Example 1 is that the pH value of the litchi pomace is not adjusted.
对比例2Comparative Example 2
本对比例与实施例1的区别为:未添加碳酸钙。The difference between this comparative example and Example 1 is that no calcium carbonate is added.
对比例3Comparative Example 3
本对比例与实施例1的区别为:所述益生菌为植物乳杆菌、乳酸片球菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的混合物。The difference between this comparative example and Example 1 is that the probiotics are a mixture of Lactobacillus plantarum, Pediococcus acidilactici, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae.
所述植物乳杆菌、乳酸片球菌、干酪乳杆菌、地衣芽孢杆菌、酿酒酵母的质量比为2:3:5:2:1。The mass ratio of the Lactobacillus plantarum, Pediococcus acidilactici, Lactobacillus casei, Bacillus licheniformis and Saccharomyces cerevisiae is 2:3:5:2:1.
对比例4Comparative Example 4
本对比例与实施例1的区别为:所述益生菌为枯草芽孢杆菌、地衣芽孢杆菌、酿酒酵母的混合物。The difference between this comparative example and Example 1 is that the probiotics are a mixture of Bacillus subtilis, Bacillus licheniformis and Saccharomyces cerevisiae.
所述枯草芽孢杆菌、地衣芽孢杆菌、酿酒酵母的质量比为2:3:4:2:1。The mass ratio of the Bacillus subtilis, Bacillus licheniformis and Saccharomyces cerevisiae is 2:3:4:2:1.
对比例5Comparative Example 5
本对比例与实施例1的区别为:所述益生菌为植物乳杆菌、乳酸片球菌、枯草芽孢杆菌、干酪乳杆菌、酿酒酵母的混合物。The difference between this comparative example and Example 1 is that the probiotics are a mixture of Lactobacillus plantarum, Pediococcus acidilactici, Bacillus subtilis, Lactobacillus casei and Saccharomyces cerevisiae.
所述枯草芽孢杆菌、干酪乳杆菌、酿酒酵母的质量比为2:3:4:5:1。The mass ratio of the bacillus subtilis, lactobacillus casei and saccharomyces cerevisiae is 2:3:4:5:1.
对比例6Comparative Example 6
本对比例与实施例1的区别为:所述复合酶为纤维素酶。The difference between this comparative example and Example 1 is that the composite enzyme is cellulase.
对比例7Comparative Example 7
本对比例与实施例1的区别为:所述复合菌和复合酶的质量比为1:1。The difference between this comparative example and Example 1 is that the mass ratio of the composite bacteria to the composite enzyme is 1:1.
对比例8Comparative Example 8
本对比例与实施例1的区别为:S3、将发酵后的荔枝果渣和饲料混合均匀,得到动物饲料。The difference between this comparative example and Example 1 is as follows: S3, the fermented litchi pomace and the feed are mixed evenly to obtain animal feed.
对比例9Comparative Example 9
本对比例与实施例1的区别为:S3、将发酵后的荔枝果渣和饲料混合均匀,再添加一定量的解脂假丝酵母,在28℃下二次发酵36h,得到动物饲料。The difference between this comparative example and Example 1 is as follows: S3, the fermented litchi pomace and feed are mixed evenly, and a certain amount of Candida lipolytica is added, and secondary fermentation is carried out at 28° C. for 36 hours to obtain animal feed.
对比例10Comparative Example 10
本对比例与实施例1的区别为:所述益生元为果寡糖。The difference between this comparative example and Example 1 is that the prebiotic is fructooligosaccharide.
性能测试Performance Testing
参考GB/T 23527-2009《蛋白酶活力》中的方法测试发酵后的荔枝果渣中蛋白酶的活性;参考龚小洁等发表的论文《乳酸菌发酵对荔枝果渣理化性质的影响》中的方法测试发酵后的荔枝果渣中可溶性酚含量。选取39头初始体重为5.5~6kg的断奶幼猪,随机分成13组,每组3头,每组对应喂养实施例1~3和对比例1~7制备的动物饲料,每天早中晚喂食三次,测量初始体重和喂养30天后猪仔的重量,取平均值,计算猪仔增重率,增重率(%)=(30天后体重-初始体重)/初始体重×100%。结果如表1所示。The activity of protease in the fermented litchi pomace was tested with reference to the method in GB/T 23527-2009 "Protease Activity"; the soluble phenol content in the fermented litchi pomace was tested with reference to the method in the paper "Effects of Lactic Acid Bacteria Fermentation on the Physical and Chemical Properties of Litchi Pomace" published by Gong Xiaojie et al. 39 weaned piglets with an initial weight of 5.5-6 kg were randomly divided into 13 groups, 3 piglets in each group, and each group was fed with animal feed prepared in Examples 1-3 and Comparative Examples 1-7 three times a day in the morning, noon and evening, and the initial weight and the weight of the piglets after 30 days of feeding were measured, and the average value was taken to calculate the weight gain rate of the piglets, weight gain rate (%) = (weight after 30 days - initial weight) / initial weight × 100%. The results are shown in Table 1.
表1测定结果Table 1 Measurement results
据统计本发明实施例1~3所制备的发酵后的荔枝果渣,其蛋白酶活性高,可溶性酚含量高,制备得到的动物饲料能够大大增加动物的体重,使得动物健康生长。对比例1未调节pH值,对比例2未添加碳酸钙,对比例3未添加枯草芽孢杆菌,对比例4未添加干酪乳杆菌,对比例5未添加地衣芽孢杆菌,对比例6未添加果胶酶,对比例7复合酶添加量过少,对比例8未进行二次发酵,对比例9未添加益生元,对比例10未添加甘露寡糖,制备得到的发酵后的荔枝果渣蛋白酶活性和可溶性酚含量均较低,动物生长的较缓慢。因此,采用本申请所述的方法提高了发酵荔枝果渣中可溶性酚等营养成分,同时提高了荔枝果渣中蛋白酶的活性,从而有利于将果渣中大分子蛋白物质分解成易吸收的小分子物质,将该发酵荔枝果渣应用于制备动物饲料,并进行二次发酵,能够进一步降解抗营养物质和纤维素等成分,提高易吸收的营养物质含量,为动物提供充足的营养,促进动物生长发育。此外,本发明提高了荔枝果渣的综合利用率,增加荔枝产业经济附加值。According to statistics, the fermented litchi pomace prepared in Examples 1 to 3 of the present invention has high protease activity and high soluble phenol content, and the prepared animal feed can greatly increase the weight of animals and make animals grow healthily. Comparative Example 1 does not adjust the pH value, Comparative Example 2 does not add calcium carbonate, Comparative Example 3 does not add Bacillus subtilis, Comparative Example 4 does not add Lactobacillus casei, Comparative Example 5 does not add Bacillus licheniformis, Comparative Example 6 does not add pectinase, Comparative Example 7 does not add too little complex enzyme, Comparative Example 8 does not perform secondary fermentation, Comparative Example 9 does not add prebiotics, Comparative Example 10 does not add manno-oligosaccharide, and the prepared fermented litchi pomace has low protease activity and soluble phenol content, and the animal grows slowly. Therefore, the method described in the present application increases the soluble phenol and other nutrients in the fermented litchi pomace, and at the same time increases the activity of the protease in the litchi pomace, thereby facilitating the decomposition of the macromolecular protein substances in the pomace into easily absorbable small molecular substances, and the fermented litchi pomace is applied to prepare animal feed, and secondary fermentation is performed, which can further degrade anti-nutrient substances and cellulose and other components, increase the content of easily absorbable nutrients, provide sufficient nutrition for animals, and promote animal growth and development. In addition, the present invention increases the comprehensive utilization rate of litchi pomace and increases the economic added value of the litchi industry.
以上所述是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明所述原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
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