Pyrus pyrifolia pomace and palm kernel meal compound fermented protein feed, and preparation method and application thereof
Patent Information
- Application Number
- CN202611142903.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-07-30
- Publication Date
- 2026-09-25
AI Technical Summary
然而，现有技术中对梨渣和棕榈粕的发酵处理多停留在单一原料、单一菌株或单一发酵方式的层面，主要存在以下不足：（1）梨渣与棕榈粕分别单独发酵，无法充分利用梨渣富含的碳水化合物与棕榈粕富含的蛋白质两者间天然营养互补优势；（2）缺乏针对梨渣与棕榈粕复配体系的专用菌株组合筛选与发酵工艺参数的系统优化；（3）发酵方式上，多采用单一的好氧或厌氧发酵，或将多种微生物同时接种，难以同步实现大分子物质降解、蛋白质有效富集与产物贮存性能改善；且多菌同时接种时易出现菌株间相互抑制，导致发酵过程不稳定、效果难以重现；（4）单一菌株发酵对原料中木质素、甘露聚糖等抗营养因子的降解能力有限，对产物蛋白质含量与生物可利用性的提升明显不足
[0024]本发明采用先接种酿酒酵母再接种贝莱斯芽孢杆菌的分阶段接种策略，突破了现有技术中多菌应同时接种的惯常做法；酿酒酵母优先代谢可快速消耗基质中残余氧气，形成有利于芽孢杆菌生长代谢的微氧环境，并释放初步降解产物供后续利用，有效避免了多菌同时接种时常见的相互抑制，所得发酵产物的真蛋白含量达到20 mg/g以上，显著高于其他接种方式，实现了菌株间的协同增效作用。
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of microbial fermentation engineering and feed processing technology, specifically relating to a compound fermented protein feed of pear pomace and palm meal, its preparation method and application. Background Technology
[0002] Pear pomace is a major byproduct of the pear processing industry, including pear juice and canned pears. my country is a major producer and processor of pears, generating a large amount of pear pomace annually. The dry basis of pear pomace contains over 60% carbohydrates and is rich in cellulose, pectin, and other polysaccharides, making it a potential carbon source for microbial fermentation. However, pear pomace has extremely low protein content, only about 2%–4%, a high proportion of stone cells, and a large molecular structure that is difficult to degrade naturally. Currently, pear pomace is mostly discarded directly or simply composted, which not only causes a serious waste of biomass resources but also easily leads to environmental problems such as spoilage and odor spread.
[0003] Palm meal is a byproduct of palm oil processing. It contains 14%–21% crude protein and is relatively rich in essential amino acids, making it a valuable and inexpensive protein feed ingredient. However, palm meal is high in crude fiber, lignin, and non-starch polysaccharides such as mannan, which are anti-nutritional factors. These components are tightly bound to protein, resulting in low digestibility and utilization by monogastric animals. Direct feeding significantly affects the absorption efficiency of nutrients in livestock and poultry, greatly limiting the effective application of palm meal in the feed industry.
[0004] Microbial solid-state fermentation is the mainstream technology for improving the quality of fruit pomace and oilseed meal by-product feed. However, the existing technologies for fermenting pear pomace and palm meal are mostly limited to single raw materials, single strains or single fermentation methods, which have the following shortcomings: (1) Pear pomace and palm meal are fermented separately, which cannot make full use of the natural nutritional complementary advantages between the carbohydrates rich in pear pomace and the proteins rich in palm meal; (2) There is a lack of screening of special strain combinations for pear pomace and palm meal compound systems and systematic optimization of fermentation process parameters; (3) In terms of fermentation methods, single aerobic or anaerobic fermentation is mostly used, or multiple microorganisms are inoculated at the same time, which makes it difficult to simultaneously achieve the degradation of macromolecules, effective enrichment of proteins and improvement of product storage performance; and when multiple bacteria are inoculated at the same time, mutual inhibition between strains is likely to occur, resulting in unstable fermentation process and difficulty in reproducing the effect; (4) The ability of single strain fermentation to degrade anti-nutritional factors such as lignin and mannan in raw materials is limited, and the improvement of product protein content and bioavailability is significantly insufficient.
[0005] Therefore, the key issue that urgently needs to be addressed is: to develop a method for preparing compound protein feed that can simultaneously leverage the complementary nutritional advantages of pear pomace and palm meal, and achieve multiple objectives such as macromolecular degradation, removal of anti-nutritional factors, protein enrichment, and improved storage stability. Summary of the Invention
[0006] To address the aforementioned technical problems, the purpose of this invention is to provide a compound fermented protein feed of pear pomace and palm meal, its preparation method, and its application.
[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:
[0008] A method for preparing a compound fermented protein feed of pear pomace and palm meal includes the following steps:
[0009] S1. Mix pear pomace and palm meal in a certain proportion, adjust the moisture content, and prepare a solid fermentation culture medium;
[0010] S2. First, inoculate the solid fermentation medium of S1 with the seed liquid of Saccharomyces cerevisiae, and after a period of aerobic solid fermentation, inoculate the seed liquid of Bacillus belye and continue aerobic solid fermentation.
[0011] S3. Inoculate the aerobic fermentation product obtained in step S2 with Lactobacillus acidophilus seed liquid and carry out anaerobic solid-state fermentation to obtain pear pomace and palm meal compound fermented protein feed.
[0012] A further improvement of the technical solution of the present invention is that the mass ratio of pear pomace to palm meal in step S1 is (6~8):(2~4).
[0013] A further improvement of the technical solution of the present invention is that the solid fermentation culture medium further includes auxiliary materials and supplementary nitrogen source; the auxiliary materials include wheat bran, and the amount of wheat bran added is 15% to 20% of the total mass of pear pomace and palm meal; the supplementary nitrogen source includes urea, and the amount of urea added is 0.5% to 2.0% of the mass of solid fermentation culture medium.
[0014] A further improvement of the technical solution of the present invention is that the dry basis moisture content of the solid fermentation culture medium is 110%~150%; the dry basis moisture content refers to the percentage of the mass of water added when preparing the culture medium to the total mass of the pear pomace, palm meal and wheat bran actually weighed.
[0015] A further improvement of the technical solution of the present invention is that, in step S2, the inoculation volume ratio of Saccharomyces cerevisiae seed liquid to Bacillus vesiculosus seed liquid is (1~3):(2~4); and, in terms of volume / mass ratio, the total inoculation volume of Saccharomyces cerevisiae seed liquid and Bacillus vesiculosus seed liquid is 10%~13% of the mass of the solid fermentation culture medium.
[0016] A further improvement of the technical solution of the present invention is that the aerobic solid-state fermentation temperature of the brewing yeast is 30~40℃ and the time is 12~36 h; the aerobic solid-state fermentation temperature of Bacillus belysin is 30~40℃ and the total aerobic solid-state fermentation time is 48~60 h.
[0017] A further improvement of the technical solution of the present invention is that, in terms of volume / mass ratio, the inoculation amount of Lactobacillus acidophilus seed liquid in step S3 is 6% to 12% of the mass of the solid fermentation culture medium, and the anaerobic solid fermentation temperature is 30 to 40°C and the time is 18 to 36 hours.
[0018] A further improvement of the technical solution of the present invention is that the brewing yeast seed liquid is prepared by inoculating brewing yeast into YPD liquid culture medium and culturing it at 30 ℃ for 24 h;
[0019] The Bacillus belysin seed culture was prepared by inoculating Bacillus belysin into LB liquid medium at an initial pH of 8, an inoculum size of 15%, and a liquid volume of 24%, and then incubating at 37 °C and 180 rpm for 24 h with shaking.
[0020] The Lactobacillus acidophilus seed culture was prepared by inoculating Lactobacillus acidophilus into MRS liquid medium and culturing it at 37 °C for 24 h.
[0021] The present invention also discloses a compound fermented protein feed of pear pomace and palm meal, wherein the pH of the compound fermented protein feed of pear pomace and palm meal is 5.5~6.5, and the true protein content is not less than 20 mg / g based on the wet weight of the fermentation product.
[0022] The present invention also discloses the application of the pear pomace and palm meal compound fermented protein feed in livestock and poultry feed.
[0023] Due to the adoption of the above technical solution, the beneficial effects of the present invention are as follows:
[0024] This invention employs a phased inoculation strategy, first inoculating with Saccharomyces cerevisiae and then with Bacillus belyssioides, which breaks through the conventional practice of inoculating multiple bacteria simultaneously in existing technologies. The preferential metabolism of Saccharomyces cerevisiae can rapidly consume the residual oxygen in the substrate, forming a micro-aerobic environment conducive to the growth and metabolism of Bacillus belyssioides, and releasing preliminary degradation products for subsequent use. This effectively avoids the mutual inhibition commonly seen when multiple bacteria are inoculated simultaneously. The true protein content of the obtained fermentation product reaches more than 20 mg / g, which is significantly higher than other inoculation methods, realizing the synergistic effect between strains.
[0025] This invention utilizes the natural nutritional complementarity of pear pomace, which is rich in carbohydrates, and palm meal, which is rich in protein. By optimizing the ratio of raw materials and the proportion of microorganisms, the aerobic solid-state fermentation product can achieve a maximum true protein content of 25.64 mg / g, a water-soluble protein content of 2.07 mg / g, and a reducing sugar content of 18.21 mg / g. This achieves effective enrichment of protein and efficient degradation of macromolecular carbohydrates, significantly improving the nutritional value and digestibility of the feed.
[0026] This invention integrates aerobic and anaerobic fermentation: in the aerobic stage, *Saccharomyces cerevisiae* and *Bacillus belye* synergistically degrade anti-nutritional factors such as crude fiber, lignin, and mannan, while accumulating microbial protein; in the anaerobic stage, *Lactobacillus acidophilus* is introduced to produce natural antibacterial substances such as organic acids, effectively inhibiting the growth of miscellaneous and putrefactive bacteria. This integrated aerobic-anaerobic process ensures that the fermentation product possesses both high nutritional value and excellent storage properties, overcoming the technical limitations of single fermentation methods that cannot achieve both.
[0027] This invention integrates liquid culture of microorganisms, aerobic solid-state fermentation, and anaerobic solid-state fermentation into a phased, intensive process system. The operation process is clear, the process conditions are controllable, and it is easy to achieve large-scale production. This invention transforms pear pomace and palm meal, which were originally inefficient and environmentally polluting, into high-value-added compound fermented protein feed. This not only expands feed resources but also reduces agricultural processing waste and environmental pollution, resulting in significant economic and ecological benefits. Detailed Implementation
[0028] The present invention will be further described below with reference to embodiments. It should be understood that the preferred embodiments described herein are for illustrative and understanding purposes only and are not intended to limit the scope of the invention.
[0029] Unless otherwise specified, all materials and reagents used in the embodiments and comparative examples of this invention are commercially available. Experimental methods not specifically described in the embodiments are generally performed under conventional conditions in the art or as recommended by the manufacturer.
[0030] The *Saccharomyces cerevisiae*, *Bacillus belyss*, and *Lactobacillus acidophilus* mentioned are all strains with corresponding fermentation activities known in the art. In this embodiment of the invention, commercially available strains were selected, specifically from the following sources: *Saccharomyces cerevisiae* was purchased from the China Industrial Microbial Culture Collection Center (CICC), accession number CICC 1421; *Bacillus belyss* was purchased from the China Industrial Microbial Culture Collection Center (CICC), accession number CICC 11068s; and *Lactobacillus acidophilus* was purchased from the China Industrial Microbial Culture Collection Center (CICC), accession number CICC 6074.
[0031] The *Saccharomyces cerevisiae* seed culture was prepared using YPD liquid medium (10 g / L yeast extract, 20 g / L peptone, 20 g / L glucose) and cultured at 30 °C for 24 h. The resulting seed culture OD... 600 It is 1.0~1.5;
[0032] The *Bacillus belyssus* seed culture was prepared using LB liquid medium (10 g / L tryptone, 5 g / L yeast extract, 10 g / L sodium chloride) and cultured at 37 °C and 180 rpm for 24 h. The resulting seed culture OD... 600 The value is 1.2~1.5 (preferably 1.36);
[0033] The Lactobacillus acidophilus seed culture was prepared using MRS liquid medium (MRS broth dry powder 52.24 g / L) and cultured at 37 ℃ and 100 rpm for 24 h. The OD of the resulting seed culture was... 600 It is 1.0~1.5.
[0034] The moisture content of the pear pomace and palm meal is no higher than 10%.
[0035] Example 1
[0036] A method for preparing a compound fermented protein feed of pear pomace and palm meal includes the following steps:
[0037] S1. Preparation of solid-state fermentation culture medium
[0038] Weigh out 10.5 g of pear pomace, 4.5 g of palm meal, 2.4 g of wheat bran and 0.20 g of urea, mix them together, and add 22.6 mL of water to prepare a solid fermentation medium with a dry basis moisture content of 130%.
[0039] S2, Aerobic Solid-State Fermentation
[0040] First, inoculate 1.8 mL of Saccharomyces cerevisiae seed culture into the solid-state fermentation medium of step S1, and ferment aerobic solid-state fermentation at 37 ℃ for 24 h. Then, inoculate 2.7 mL of Bacillus belye seed culture and continue aerobic solid-state fermentation at 37 ℃ for 24 h.
[0041] S3, Anaerobic Solid-State Fermentation
[0042] Add 3.6 mL of Lactobacillus acidophilus seed liquid to the aerobic fermentation product obtained in step S2, and carry out anaerobic solid-state fermentation at 37 ℃ for 24 h to obtain pear pomace and palm meal compound fermented protein feed.
[0043] Example 2
[0044] A method for preparing a compound fermented protein feed of pear pomace and palm meal includes the following steps:
[0045] S1. Preparation of solid-state fermentation culture medium
[0046] Weigh out 10.0 g of pear pomace, 5.0 g of palm meal, 2.25 g of wheat bran and 0.36 g of urea, mix them together, and add 19.0 mL of water to prepare a solid fermentation medium with a dry basis moisture content of 110%.
[0047] S2, Aerobic Solid-State Fermentation
[0048] First, inoculate 1.2 mL of Saccharomyces cerevisiae seed culture into the solid-state fermentation medium of step S1, and ferment aerobic solid-state fermentation at 30 ℃ for 36 h. Then, inoculate 2.4 mL of Bacillus belye seed culture and continue aerobic solid-state fermentation at 30 ℃ for 24 h.
[0049] S3, Anaerobic Solid-State Fermentation
[0050] 2.2 mL of Lactobacillus acidophilus seed liquid was added to the aerobic fermentation product obtained in step S2, and anaerobic solid-state fermentation was carried out at 30 °C for 36 h to obtain a compound fermented protein feed of pear pomace and palm meal.
[0051] Example 3
[0052] A method for preparing a compound fermented protein feed of pear pomace and palm meal includes the following steps:
[0053] S1. Preparation of solid-state fermentation culture medium
[0054] Weigh out 12.0 g of pear pomace, 3.0 g of palm meal, 3.0 g of wheat bran and 0.90 g of urea, mix them together, and add 27.0 mL of water to prepare a solid fermentation medium with a dry basis moisture content of 150%.
[0055] S2, Aerobic Solid-State Fermentation
[0056] First, inoculate 2.5 mL of Saccharomyces cerevisiae seed culture into the solid-state fermentation medium of step S1, and ferment aerobic solid-state fermentation at 40 ℃ for 12 h. Then, inoculate 3.4 mL of Bacillus belye seed culture and continue aerobic solid-state fermentation at 40 ℃ for 36 h.
[0057] S3, Anaerobic Solid-State Fermentation
[0058] Add 5.5 mL of Lactobacillus acidophilus seed liquid to the aerobic fermentation product obtained in step S2, and carry out anaerobic solid-state fermentation at 40 °C for 18 h to obtain pear pomace and palm meal compound fermented protein feed.
[0059] Example 4
[0060] A method for preparing a compound fermented protein feed of pear pomace and palm meal includes the following steps:
[0061] S1. Preparation of solid-state fermentation culture medium
[0062] Weigh out 9.0 g of pear pomace, 6.0 g of palm meal, 2.4 g of wheat bran and 0.20 g of urea, mix them together, and add 22.6 mL of water to prepare a solid fermentation medium with a dry basis moisture content of 130%.
[0063] S2, Aerobic Solid-State Fermentation
[0064] First, inoculate 2.7 mL of Saccharomyces cerevisiae seed culture into the solid-state fermentation medium of step S1, and ferment aerobic solid-state fermentation at 35 ℃ for 30 h. Then, inoculate 1.8 mL of Bacillus belye seed culture and continue aerobic solid-state fermentation at 37 ℃ for 24 h.
[0065] S3, Anaerobic Solid-State Fermentation
[0066] Add 3.6 mL of Lactobacillus acidophilus seed liquid to the aerobic fermentation product obtained in step S2, and carry out anaerobic solid-state fermentation at 37 ℃ for 24 h to obtain pear pomace and palm meal compound fermented protein feed.
[0067] Comparative Example 1
[0068] A method for preparing a compound fermented protein feed of pear pomace and palm meal differs from Example 1 in that: in step S2, the aerobic solid-state fermentation stage first involves fermentation with Bacillus belye, followed by fermentation with Saccharomyces cerevisiae, i.e.:
[0069] First, inoculate 2.7 mL of Bacillus belye seed culture into the solid-state fermentation medium of step S1, and ferment aerobic solid-state fermentation at 37 ℃ for 24 h. Then, inoculate 1.8 mL of Saccharomyces cerevisiae seed culture and continue aerobic solid-state fermentation at 37 ℃ for 24 h.
[0070] Comparative Example 2
[0071] A method for preparing a compound fermented protein feed of pear pomace and palm meal differs from Example 1 in that: in step S2, the aerobic solid-state fermentation stage simultaneously carries out fermentation of Bacillus berberis and Saccharomyces cerevisiae, that is:
[0072] Simultaneously inoculate 1.8 mL of Saccharomyces cerevisiae seed culture and 2.7 mL of Bacillus belye seed culture into the solid-state fermentation medium of step S1, and ferment aerobic solid-state fermentation at 37°C for 24 h.
[0073] Product physicochemical property testing:
[0074] True protein was determined using the Kjeldahl method (the sample was first precipitated with 10% trichloroacetic acid solution to remove non-protein nitrogen, and then determined according to GB5009.5 Kjeldahl method).
[0075] Water-soluble proteins were determined using the Coomassie Brilliant Blue (Bradford) colorimetric method.
[0076] The reducing sugar content was determined by the 3,5-dinitrosalicylic acid (DNS) colorimetric method.
[0077] The aerobic fermentation products and final fermentation products of Examples 1-4 and Comparative Examples 1-2 were tested for physicochemical properties. The specific results are as follows:
[0078]
[0079] The pear pomace and palm meal compound fermented protein feed prepared by this invention has a pH of 5.5-6.5 and a true protein content of not less than 20 mg / g based on the wet weight of the fermentation product, making it suitable for use in livestock and poultry feed.
[0080] The present invention has been described in detail with reference to the above embodiments. It should be noted that the above embodiments are merely illustrative examples. Without departing from the spirit and essence of the present invention, those skilled in the art can devise various alternatives and improvements to the present invention, all of which should be understood to be within the scope of protection of the present invention.
Claims
1. A method for preparing a compound fermented protein feed of pear pomace and palm meal, characterized in that, Includes the following steps: S1. Mix pear pomace and palm meal in a certain proportion, adjust the moisture content, and prepare a solid fermentation culture medium; S2. First, inoculate the solid fermentation medium of step S1 with the seed liquid of Saccharomyces cerevisiae, and after a period of aerobic solid fermentation, inoculate the seed liquid of Bacillus belye and continue aerobic solid fermentation. S3. Inoculate the aerobic fermentation product obtained in step S2 with Lactobacillus acidophilus seed liquid and carry out anaerobic solid-state fermentation to obtain pear pomace and palm meal compound fermented protein feed.
2. The method for preparing a compound fermented protein feed of pear pomace and palm meal according to claim 1, characterized in that, In step S1, the mass ratio of pear pomace to palm meal is (6~8):(2~4).
3. The method for preparing a compound fermented protein feed of pear pomace and palm meal according to claim 2, characterized in that, The solid-state fermentation medium also includes auxiliary materials and a supplementary nitrogen source; the auxiliary materials include wheat bran, and the amount of wheat bran added is 15% to 20% of the total mass of pear pomace and palm meal; the supplementary nitrogen source includes urea, and the amount of urea added is 0.5% to 2.0% of the mass of the solid-state fermentation medium.
4. The method for preparing a compound fermented protein feed of pear pomace and palm meal according to claim 3, characterized in that, The dry basis moisture content of the solid fermentation medium is 110%~150%.
5. The method for preparing a compound fermented protein feed of pear pomace and palm meal according to claim 1, characterized in that, In step S2, the inoculation volume ratio of Saccharomyces cerevisiae seed liquid to Bacillus vesiculus seed liquid is (1~3):(2~4); the total inoculation amount of Saccharomyces cerevisiae seed liquid and Bacillus vesiculus seed liquid is 10%~13% of the mass of solid fermentation medium, based on the volume / mass ratio.
6. The method for preparing a compound fermented protein feed of pear pomace and palm meal according to claim 5, characterized in that, The aerobic solid-state fermentation temperature of the brewing yeast is 30~40 ℃ and the time is 12~36 h; the aerobic solid-state fermentation temperature of Bacillus belysin is 30~40 ℃ and the total aerobic solid-state fermentation time is 48~60 h.
7. A method for preparing a compound fermented protein feed of pear pomace and palm meal according to any one of claims 1 to 6, characterized in that, In step S3, the inoculation amount of Lactobacillus acidophilus seed liquid is 6% to 12% of the mass of the solid fermentation medium, and the anaerobic solid fermentation temperature is 30 to 40 °C and the time is 18 to 36 h.
8. The method for preparing a compound fermented protein feed of pear pomace and palm meal according to claim 7, characterized in that, The Saccharomyces cerevisiae seed culture was prepared by inoculating Saccharomyces cerevisiae into YPD liquid medium and culturing it at 30 ℃ for 24 h. The Bacillus belysin seed culture was prepared by inoculating Bacillus belysin into LB liquid medium and incubating it at 37 °C and 180 rpm for 24 h under the conditions of initial pH 8, inoculum size 15%, and liquid volume 24%. The Lactobacillus acidophilus seed culture was prepared by inoculating Lactobacillus acidophilus into MRS liquid medium and culturing it at 37 °C for 24 h.
9. A pear pomace and palm meal fermented protein feed prepared by the method described in any one of claims 1 to 8, characterized in that, The pH of the pear pomace and palm meal compound fermented protein feed is 5.5~6.5, and the true protein content is not less than 20 mg / g based on the wet weight of the fermentation product.
10. The application of the pear pomace and palm meal compound fermented protein feed as described in claim 9 in livestock and poultry feed.