A fermented palm meal with high organic acid content and its preparation process

Abstract

This invention relates to the field of biofeed technology, specifically disclosing a high-organic-acid fermented palm meal and its preparation process. The preparation process mainly includes the following steps: (1) mixing a compound microbial fermentation agent, a compound enzyme preparation, and water to prepare a seed solution; (2) mixing a certain amount of palm meal, food yeast powder, wheat bran, and water with the seed solution from step (1); (3) fermenting the uniformly mixed palm meal; and (4) drying the fermented material to obtain fermented palm meal. The palm meal obtained by this preparation process contains a variety of organic acids, with an organic acid content ≥8wt%, significantly improving the digestibility of palm meal by animals and also increasing the utilization efficiency of raw materials.

Description

Technical Field

[0001] This invention belongs to the field of biofeed technology, specifically relating to a high-organic-acid fermented palm meal and its preparation process. Background Technology

[0002] Palm meal is the residue left after palm kernels are pressed and the palm oil is removed. Currently, Indonesia and Malaysia are the world's largest producers and exporters of palm meal, with huge production volumes and low prices. It has been used as a common feed ingredient abroad for many years. In recent years, due to the shortage and price increase of conventional feed ingredients, palm meal has gradually been introduced into my country for application.

[0003] Palm meal has a fine texture and a slight chocolate and alcoholic aroma. Its quality varies depending on the variety, degree of hulling, harvest time, and processing method. Rich in linoleic acid, vitamins B and E, and trace elements such as phosphorus, manganese, and iron, palm meal is palatable and has high feed value, making it suitable for aquaculture and various livestock farming. Although nutritious and relatively inexpensive, its high crude fiber content, sandy appearance, and abundant polysaccharide structure result in low digestibility and utilization by monogastric animals, limiting its use in animal husbandry. Summary of the Invention

[0004] The technical problem solved by this invention is that palm meal contains a large amount of insoluble mannan and crude fiber, has a sandy appearance and a large amount of polysaccharide structure, which makes it difficult for monogastric animals to digest and utilize it, thus limiting the use of palm meal in animal husbandry.

[0005] To address these issues, the present invention aims to provide a method for preparing fermented palm meal, which uses a combination of enzymatic hydrolysis and biological fermentation to convert the difficult-to-utilize mannans or cellulose in palm meal into organic acids, thereby improving the digestibility of palm meal for animals.

[0006] Specifically, the present invention proposes the following technical solution.

[0007] The purpose of this invention is to provide a high-organic-acid fermented palm meal, characterized in that the palm meal contains ≥15wt% crude protein and ≥8wt% total organic acids; preferably, the palm meal contains 15wt%–30wt% crude protein and 8wt%–20wt% total organic acids; preferably, the palm meal contains 15wt%–20wt% crude protein and 8wt%–12wt% total organic acids; preferably, the palm meal contains 15wt%–17wt% crude protein and 8wt%–9wt% total organic acids.

[0008] Preferably, the acid-soluble protein content in the crude protein is ≥25wt%; more preferably 25wt% to 35wt%, and even more preferably 31wt% to 35wt%.

[0009] Preferably, the organic acid contains lactic acid, acetic acid, citric acid, malic acid, and succinic acid.

[0010] Preferably, the organic acid contains 3 wt% to 5 wt% lactic acid, 0.1 wt% to 1 wt% acetic acid, 2.5 wt% to 4 wt% citric acid, 0.1 wt% to 1 wt% malic acid, and 0.06 wt% to 1 wt% succinic acid; more preferably, the organic acid contains 3.5 wt% to 4.5 wt% lactic acid, 0.1 wt% to 0.6 wt% acetic acid, 3 wt% to 4 wt% citric acid, 0.1 wt% to 0.8 wt% malic acid, and 0.06 wt% to 0.5 wt% succinic acid; even more preferably, the organic acid contains 3.9 wt% to 4.3 wt% lactic acid, 0.1 wt% to 0.4 wt% acetic acid, 3.1 wt% to 3.9 wt% citric acid, 0.1 wt% to 0.5 wt% malic acid, and 0.06 wt% to 0.3 wt% succinic acid.

[0011] Another object of the present invention is to provide a method for preparing the above-mentioned high organic acid fermented palm meal, characterized in that it includes the following steps:

[0012] (1) Preparation of seed liquid: Take 0.3 to 15 parts by weight of compound microbial fermentation agent and 0.4 to 20 parts by weight of compound enzyme preparation and mix them evenly with water to prepare seed liquid;

[0013] (2) Inoculation and mixing: Take a certain amount of palm meal, food yeast powder, wheat bran, water and mix them evenly with the seed liquid in step (1);

[0014] (3) Ferment the well-mixed palm meal; and

[0015] (4) Dry the material after fermentation to obtain fermented palm meal.

[0016] Preferably, in step (1), the seed liquid contains 0.3 to 11 parts by weight of compound microbial fermentation agent and 0.4 to 16 parts by weight of compound enzyme preparation; more preferably, the seed liquid contains 6.5 to 11 parts by weight of compound microbial fermentation agent and 7 to 12 parts by weight of compound enzyme preparation.

[0017] Preferably, in step (1), the compound microbial fermentation agent is selected from one or more of yeast, Bacillus and lactic acid bacteria; preferably, the compound microbial fermentation agent contains yeast, Bacillus and lactic acid bacteria; more preferably, the compound microbial fermentation agent contains 0.1 to 5 parts of yeast, 0.1 to 5 parts of Bacillus and 0.1 to 5 parts of lactic acid bacteria; more preferably, the compound microbial fermentation agent contains 1 to 5 parts of yeast, 0.5 to 2 parts of Bacillus and 4 to 5 parts of lactic acid bacteria.

[0018] Preferably, the yeast is selected from one or more of Saccharomyces cerevisiae, Candida utilis, and Kluyveromyces oryzae.

[0019] Preferably, the yeast contains Saccharomyces cerevisiae and Kluyveromyces oryzae;

[0020] More preferably, the yeast contains 0.05 to 4 parts by weight of Saccharomyces cerevisiae and / or 0.05 to 4 parts by weight of Kluyveromyces oryzae; more preferably, the yeast contains 0.5 to 2.5 parts by weight of Saccharomyces cerevisiae and / or 0.5 to 2.5 parts by weight of Kluyveromyces oryzae.

[0021] Preferably, the Kluyveromycin is Kluyveromycin KW-6.

[0022] Preferably, the Bacillus is selected from one or more of Bacillus subtilis, Bacillus licheniformis, Bacillus coagulans, Bacillus pumilus, and Bacillus laterosporus.

[0023] Preferably, the Bacillus species includes Bacillus pumilus and / or Bacillus coagulans.

[0024] Preferably, the Bacillus contains 0.1 to 1 part by weight of Bacillus subtilis, 0.1 to 1 part by weight of Bacillus licheniformis, 0.1 to 1 part by weight of Bacillus coagulans, 0.1 to 1 part by weight of Bacillus pumilus, and / or 0.1 to 1 part by weight of Bacillus laterosporus.

[0025] More preferably, the Bacillus contains 0.1 to 1 part by weight of Bacillus subtilis, 0.5 to 1 part by weight of Bacillus licheniformis, 0.1 to 1 part by weight of Bacillus coagulans, 0.5 to 1 part by weight of Bacillus pumilus, and / or 0.1 to 1 part by weight of Bacillus laterosporus.

[0026] More preferably, the Bacillus is 0.1 to 1 part by weight of Bacillus coagulans and 0.5 to 1 part by weight of Bacillus pumilus.

[0027] Preferably, the lactic acid bacteria are selected from one or more of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus acidophilus, Enterococcus faecalis, and Enterococcus faecium;

[0028] Preferably, the lactic acid bacteria contain Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus, Enterococcus faecalis, and Enterococcus faecium;

[0029] More preferably, the lactic acid bacteria contain 0.5 to 1.5 parts by weight of Lactobacillus plantarum, 0.5 to 1.5 parts by weight of Lactobacillus casei, 0.5 to 1.5 parts by weight of Lactobacillus acidophilus, 0.5 to 1.5 parts by weight of Enterococcus faecalis, and / or 0.5 to 1.5 parts by weight of Enterococcus faecium.

[0030] More preferably, the lactic acid bacteria contain 1 to 1.5 parts by weight of Lactobacillus plantarum, 1 to 1.5 parts by weight of Lactobacillus casei, 0.5 to 1 part by weight of Lactobacillus acidophilus, 0.5 to 1 part by weight of Enterococcus faecalis and 0.5 to 1 part by weight of Enterococcus faecium.

[0031] Preferably, in step (1), the complex enzyme preparation is selected from one or more of protease, mannanase, cellulase and dextranase;

[0032] Preferably, the compound enzyme preparation contains protease, mannanase, cellulase and dextranase.

[0033] More preferably, the compound enzyme preparation contains 0.1 to 5 parts by weight of protease, 0.1 to 5 parts by weight of mannanase, 0.1 to 5 parts by weight of cellulase and 0.1 to 5 parts by weight of glucanase;

[0034] More preferably, the compound enzyme preparation contains 4-5 parts by weight of protease, 0.5-1.5 parts by weight of mannanase, 4-5 parts by weight of cellulase and 0.1-1 parts by weight of glucanase.

[0035] Preferably, in the compound enzyme preparation, the protease activity is 50,000 to 300,000 U / g, the mannanase activity is 5,000 to 100,000 U / g, the cellulase activity is 5,000 to 100,000 U / g, and / or the dextranase activity is 5,000 to 100,000 U / g.

[0036] Preferably, in step (1), the weight of the water is 5 to 30 parts by weight; more preferably, the weight of the water is 5 to 15 parts by weight.

[0037] Preferably, in step (2), the palm meal is 50-90 parts by weight and the food yeast powder is 10-20 parts by weight; more preferably, the bran is 5-30 parts by weight and the water is 30-50 parts by weight; more preferably, the palm meal is 75-85 parts by weight, the food yeast powder is 10-20 parts by weight, the bran is 5-10 parts by weight and the water is 35-45 parts by weight.

[0038] Preferably, the fermentation method is anaerobic fermentation; more preferably, it is anaerobic fermentation in a fermentation box, anaerobic fermentation in a fermentation tank, or anaerobic fermentation in a breathing bag.

[0039] Preferably, in step (3), the fermentation temperature is 25-40°C; more preferably, the fermentation temperature is 35-40°C.

[0040] Preferably, in step (3), the fermentation time is 2 to 7 days; more preferably, the fermentation time is 4 to 6 days.

[0041] Preferably, the fermented palm meal obtained by the preparation method has a crude protein content of ≥15wt% and a total content of various organic acids of ≥8wt%; wherein, acid-soluble protein accounts for ≥25wt% of the crude protein content.

[0042] Preferably, a feed is made from the fermented palm meal or fermented palm meal obtained by the preparation method.

[0043] Preferably, the fermented palm meal or the fermented palm meal obtained by the preparation method is used in feed.

[0044] The beneficial effects of this invention compared to the prior art are as follows:

[0045] (1) Through the combination of enzyme preparation and microbial fermentation, the fermented palm meal contains a variety of organic acids, mainly lactic acid, acetic acid, malic acid, citric acid and succinic acid. The content of the five organic acids is as high as 8wt% to 9wt%, and the proportion of acid-soluble protein in crude protein can reach 25wt% to 35wt%.

[0046] (2) The fermented palm meal obtained by this invention was measured by a monogastric animal bionic digestion instrument, and the dry matter digestibility (duck) increased from 14.03% to about 35%.

[0047] (3) The results of in vitro digestion simulation of ruminants showed that the digestibility of the fermented palm meal prepared by the present invention increased from 35.46% to about 75%, an increase of 111.5%; it greatly improved the digestibility of palm meal by animals and also improved the utilization efficiency of raw materials.

[0048] Information on strain preservation:

[0049] The Kluyveromyces marxious KW-6 used in this invention was deposited at the China Center for Type Culture Collection on June 2, 2020, with accession number CCTCC NO: M 2020173. The deposit address is: Wuhan University, Wuhan, China, postal code: 430072; telephone: (027)-68754052. Detailed Implementation

[0050] As described above, the purpose of this invention is to provide a method for preparing high-organic-acid fermented palm meal, which converts the insoluble polysaccharides in palm meal into higher contents of crude protein, acid-soluble protein and organic acids, thereby significantly improving the digestibility of palm meal by animals and also improving the utilization efficiency of raw materials.

[0051] In one specific embodiment of the present invention, a fermented palm meal with high organic acid content is provided, wherein the fermented palm meal contains ≥15wt% crude protein and ≥8wt% of various organic acids; wherein acid-soluble protein accounts for ≥25% of the crude protein content.

[0052] Preferably, the organic acid contains 3.9 wt% to 4.3 wt% lactic acid, 0.1 wt% to 0.4 wt% acetic acid, 3.1 wt% to 3.9 wt% citric acid, 0.1 wt% to 0.5 wt% malic acid, and 0.06 wt% to 0.3 wt% succinic acid.

[0053] In another specific embodiment of the present invention, a method for preparing fermented palm meal with high organic acid content is provided, comprising the following steps:

[0054] (1) Preparation of seed liquid: Take 0.3 to 15 parts by weight of compound microbial fermentation agent and 0.4 to 20 parts by weight of compound enzyme preparation and mix them evenly with water to prepare seed liquid;

[0055] (2) Inoculation and mixing: Take palm meal, food yeast powder, wheat bran, water and mix them evenly with the seed liquid in step (1);

[0056] (3) Ferment the well-mixed palm meal;

[0057] (4) Dry the material after fermentation to obtain fermented palm meal.

[0058] In another specific embodiment of the present invention, a method for preparing fermented palm meal with high organic acid content is provided, comprising the following steps:

[0059] (1) Preparation of seed liquid: Take 0.3-15 parts by weight of compound microbial fermentation agent, 0.4-20 parts by weight of compound enzyme preparation and 5-30 parts by weight of water and mix them evenly to prepare seed liquid;

[0060] (2) Inoculation and mixing: Take 50-90 parts by weight of palm meal, 10-20 parts by weight of food yeast powder, 5-30 parts by weight of wheat bran, 30-50 parts by weight of water and mix them evenly with the seed liquid in step (1).

[0061] (3) Ferment the evenly mixed palm meal in an anaerobic fermentation manner, at a temperature of 25-40℃ for 2-7 days.

[0062] (4) Dry the material after fermentation to obtain fermented palm meal.

[0063] Preferably, the compound microbial fermentation agent contains 0.1-5 parts of yeast, 0.1-5 parts of Bacillus, and 0.1-5 parts of lactic acid bacteria. Preferably, the yeast contains 0.05-4 parts by weight of *Saccharomyces cerevisiae* and / or 0.05-4 parts by weight of *Kluyveromyces kwoniophylla* KW-6; more preferably, the Bacillus contains 0.1-1 parts by weight of *Bacillus subtilis*, 0.1-1 parts by weight of *Bacillus licheniformis*, 0.1-1 parts by weight of *Bacillus coagulans*, 0.1-1 parts by weight of *Bacillus pumilus*, and / or 0.1-1 parts by weight of *Bacillus laterosporus*; even more preferably, the lactic acid bacteria contain 0.5-1.5 parts by weight of *Lactobacillus plantarum*, 0.5-1.5 parts by weight of *Lactobacillus casei*, 0.5-1.5 parts by weight of *Lactobacillus acidophilus*, 0.5-1.5 parts by weight of *Enterococcus faecalis*, and / or 0.5-1.5 parts by weight of *Enterococcus faecium*.

[0064] Preferably, the compound enzyme preparation contains 0.1 to 5 parts by weight of protease, 0.1 to 5 parts by weight of mannanase, 0.1 to 5 parts by weight of cellulase and 0.1 to 5 parts by weight of glucanase.

[0065] Kluyveromyces marxious KW-6 is a strain of Kluyveromyces yeast bred by Angel Yeast Co., Ltd. This strain was isolated and purified from fresh yak milk obtained from herders in Shannan City, Tibet. Morphological observation under a high-power microscope revealed that the yeast strain is slender and oval, primarily budding at both ends and reproducing asexually. After culturing on solid culture plates at 28°C for 10 hours, the colonies were round, white, raised, transparent, with a moist and smooth surface and neat edges. Its food-grade safety attributes were verified through routine morphological observation, physiological and biochemical tests, and toxicological evaluation. Then, using PCR technology, the 26S rDNA D1 / D2 region gene sequence was identified, confirming that the strain of this invention belongs to the genus Kluyveromyces marxious. It was deposited at the China Center for Type Culture Collection on June 2, 2020, with accession number CCTCC NO: M 2020173.

[0066] Experimental methods not specifically described in the following examples were generally performed under standard conditions or as recommended by the manufacturer. All raw materials used in the examples are commercially available products unless otherwise stated. The sources of the reagents and instruments used in the examples are as follows:

[0067] Table 1. Reagents / instruments and manufacturers used in the examples.

[0068]

[0069]

[0070] In this invention, the determination of moisture content in fermented palm meal is carried out in accordance with the People's Republic of China National Standard GB / T6435-2014: Determination of Moisture Content in Feed.

[0071] In this invention, the determination of crude protein content in fermented palm meal is based on the People's Republic of China National Standard GB / T 24318-2009: Determination of Total Nitrogen Content and Calculation of Crude Protein in Feed Raw Materials by Dumas Combustion Method; the calculation formula for crude protein content is: w(P)=F×w(N), where w(P) is the crude protein content (%) in the sample; F is the protein factor, the coefficient for converting nitrogen to protein; and w(N) is the total nitrogen content (%) in the sample.

[0072] In this invention, the determination of acid-soluble protein content in fermented palm meal is based on GB / T 22492-2008 Detection of acid-soluble protein in soybean peptide powder.

[0073] In this invention, the determination of lactic acid and citric acid content in fermented palm meal is carried out in accordance with the People's Republic of China National Standard GB / T 23877-2009: Determination of citric acid, fumaric acid and lactic acid in feed acidifiers by high performance liquid chromatography.

[0074] In this invention, the determination of malic acid content in fermented palm meal is based on the People's Republic of China National Standard GB25544-2010: Food Additive DL-Malic Acid.

[0075] In this invention, the determination of succinic acid content in fermented palm meal is based on the People's Republic of China National Standard GB29939-2013: Disodium succinate, a food additive.

[0076] In this invention, the acetic acid content in fermented palm meal was determined using high-performance liquid chromatography (HPLC). Chromatographic conditions: Analytical column: BIO-RAD Aminex HPX-87H (300mm × 7.8mm) (with guard column); mobile phase: 0.005 mol / L H₂SO₄ solution; flow rate: 0.6 mL / min; column temperature: 65℃; after equilibration and stabilization of the mobile phase, the sample was injected at a volume of 20 μL. Sample preparation: The sample was mixed with an appropriate amount of pure water in a sampling container. A suitable amount of the mixed sample was centrifuged at 4000 rpm for 10 minutes. A precise mass of the supernatant was transferred to a volumetric flask, diluted to volume with pure water, and diluted appropriately to ensure the concentration was within the standard curve range. The diluted solution was filtered through a 0.45 μm filter membrane before testing. The formula for calculating the acetic acid content is:

[0077]

[0078] Where X is the content of a certain component in the sample (μg / g); c is the content of a certain component in the sample solution (μg / mL); V is the final volume of the sample (mL); m is the mass of the sample (g); and f is the dilution factor of the sample.

[0079] In this invention, the dry matter digestibility (duck) of fermented palm meal was determined using a SDS-III monogastric animal bionic digester. The detection method was as follows: the fermented palm meal sample was crushed, passed through a 60-mesh sieve, and 1g of sample was weighed. The dry matter digestibility of the sample was obtained by operating the third-generation monogastric animal bionic digestion system according to the operating procedures (specific operation refers to the equipment operation manual provided by Hubei Zhongben Intelligent). The formula for calculating the dry matter digestibility is: Dry matter digestibility = (dry matter weight of sample before digestion - dry matter weight of sample after digestion) / dry matter weight of sample before digestion × 100%.

[0080] In this invention, the in vitro digestion assay of fermented palm meal ruminant is performed using the AGRS-Ⅲ ruminant biomimetic digester. The detection method is as follows: the fermented palm meal sample is crushed, passed through a 60-mesh sieve, and 1g of sample is weighed. The dry matter digestibility of the sample is obtained by operating the AGRS-Ⅲ ruminant biomimetic digester according to the operating procedures (specific operation refers to the equipment operation manual provided by Hubei Jingboxiang Xingwang). The formula for calculating the dry matter digestibility is: Dry matter digestibility = (Dry matter weight of sample before digestion - Dry matter weight of sample after digestion) / Dry matter weight of sample before digestion × 100%.

[0081] The present invention will be described in detail below with reference to specific embodiments:

[0082] Example 1

[0083] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 0.1 parts of Saccharomyces cerevisiae powder, 0.1 parts of Bacillus subtilis powder and 0.1 parts of Lactobacillus plantarum powder; the compound enzyme preparation contains 0.1 parts of protease, 0.1 parts of mannanase, 0.1 parts of cellulase and 0.1 parts of glucanase; mix the compound microbial fermentation agent, compound enzyme preparation and 10 parts of water and stir for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0084] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0085] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 25°C for 7 days to end the fermentation.

[0086] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0087] Example 2

[0088] (1) Preparation of seed liquid: By weight, the compound microbial fermentation agent contains 0.05 parts of Saccharomyces cerevisiae powder, 0.05 parts of Kluyveromyces kw-6 powder, 0.5 parts of Bacillus licheniformis powder, 0.5 parts of Bacillus laterosporus powder, 0.5 parts of Bacillus coagulans powder, 0.5 parts of Lactobacillus plantarum powder and 0.5 parts of Lactobacillus casei powder; the compound enzyme preparation contains 0.1 parts of protease, 1 part of mannanase, 1 part of cellulase and 1 part of glucanase; the compound microbial fermentation agent, the compound enzyme preparation and 10 parts of water are mixed and stirred for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0089] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0090] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 30℃ for 5 days to end the fermentation.

[0091] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0092] Example 3

[0093] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 0.1 parts of Kluyveromyces KW-6 powder, 1 part of Bacillus subtilis powder, 1 part of Bacillus licheniformis powder, 1 part of Bacillus coagulans powder, 1 part of Lactobacillus plantarum powder, 1 part of Lactobacillus casei powder, 1 part of Lactobacillus paracasei powder, 1 part of Enterococcus faecalis and 1 part of Enterococcus faecium; the compound enzyme preparation contains 0.1 parts of protease, 5 parts of mannanase, 5 parts of cellulase and 5 parts of glucanase; the compound microbial fermentation agent, the compound enzyme preparation and 10 parts of water are mixed and stirred for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0094] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0095] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 35°C for 3 days to end the fermentation.

[0096] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0097] Example 4

[0098] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 1 part of Saccharomyces cerevisiae powder, 0.1 part of Bacillus coagulans powder and 0.1 part of Lactobacillus casei powder; the compound enzyme preparation contains 1 part of protease, 1 part of mannanase, 0.1 part of cellulase and 5 parts of glucanase; mix the compound microbial fermentation agent, compound enzyme preparation and 10 parts of water and stir for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0099] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0100] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 40℃ for 2 days to end the fermentation.

[0101] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0102] Example 5

[0103] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 0.5 parts of Saccharomyces cerevisiae powder, 0.5 parts of Kluyveromyces kW-6 powder, 0.5 parts of Bacillus pumilus powder, 1 part of Lactobacillus plantarum powder, 1 part of Lactobacillus casei powder, 1 part of Lactobacillus paracasei powder, 1 part of Lactobacillus acidophilus powder and 1 part of Enterococcus faecalis powder; the compound enzyme preparation contains 1 part of protease, 1 part of mannanase, 0.1 part of cellulase and 5 parts of glucanase; the compound microbial fermentation agent, the compound enzyme preparation and 10 parts of water are mixed and stirred for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0104] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0105] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 35°C for 3 days to end the fermentation.

[0106] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0107] Example 6

[0108] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 1 part Kluyveromyces KW-6 bacterial powder, 1 part Bacillus subtilis bacterial powder, 1 part Bacillus licheniformis bacterial powder, 1 part Bacillus pumilus bacterial powder, 1 part Bacillus coagulans bacterial powder, 0.5 parts Enterococcus faecalis and 0.5 parts Enterococcus faecium; the compound enzyme preparation contains 1 part protease, 5 parts mannanase, 1 part cellulase and 0.1 part glucanase; the compound microbial fermentation agent, the compound enzyme preparation and 10 parts water are mixed and stirred for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0109] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0110] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 30℃ for 5 days to end the fermentation.

[0111] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0112] Example 7

[0113] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 1 part of Saccharomyces cerevisiae powder, 4 parts of Kluyveromyces kW-6 powder, 0.1 part of Bacillus laterosporus powder, 0.5 parts of Lactobacillus casei powder and 0.5 parts of Lactobacillus paracasei powder, and the compound enzyme preparation contains 5 parts of protease, 0.1 parts of mannanase, 1 part of cellulase and 5 parts of glucanase; the compound microbial fermentation agent, the compound enzyme preparation and 10 parts of water are mixed and stirred for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0114] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0115] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 25°C for 7 days to end the fermentation.

[0116] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0117] Example 8

[0118] (1) Seed liquid preparation: By weight, the compound microbial fermentation agent contains 2.5 parts of Saccharomyces cerevisiae powder, 2.5 parts of Kluyveromyces kW-6 powder, 0.5 parts of Bacillus pumilus powder, 0.5 parts of Bacillus coagulans powder, 1 part of Lactobacillus plantarum, 1 part of Lactobacillus casei, 1 part of Lactobacillus acidophilus, 1 part of Enterococcus faecalis and 1 part of Enterococcus faecium; the compound enzyme preparation contains 5 parts of protease, 1 part of mannanase, 5 parts of cellulase and 0.1 part of glucanase; the compound microbial fermentation agent, the compound enzyme preparation and 10 parts of water are mixed and stirred for 10 min at a temperature of 30±5℃ to prepare the seed liquid for later use.

[0119] (2) Inoculation and mixing: Weigh 80 parts palm meal, 15 parts food yeast powder and 5 parts wheat bran, measure 40 parts water, and mix them thoroughly with the seed liquid prepared in step (1) in a mixer.

[0120] (3) Fermentation: Transfer the evenly mixed palm meal into a fermentation bag with a breather buckle, seal it, and ferment at 30℃ for 5 days to end the fermentation.

[0121] (4) Drying: Dry the material after fermentation to prepare fermented palm meal.

[0122] First, the crude protein and acid-soluble protein content in the fermented palm meal products prepared in each embodiment were tested, as shown in the table below:

[0123] Table 2. Results of determination of crude protein and acid-soluble protein content in fermented palm meal product.

[0124]

[0125]

[0126] As can be seen from the table above, the fermented palm meal prepared in Examples 1-8 of this invention has a significantly higher acid-soluble protein content than the initial palm meal substrate, wherein the acid-soluble protein accounts for more than 25 wt% of the crude protein content.

[0127] Next, the organic acid content and dry matter digestibility of the fermented palm meal prepared in each example were tested, as shown below:

[0128] Table 3. Results of determination of organic acid and dry matter digestibility of fermented palm meal product.

[0129]

[0130] As can be seen from the table above, the fermented palm meal prepared in Examples 1-8 of this invention exhibits significantly increased contents of lactic acid, acetic acid, malic acid, citric acid, and succinic acid. Lactic acid and citric acid are particularly significantly increased, with the total content of the five organic acids obtained in this invention exceeding 8%, while the organic acid content of palm meal without microbial and enzymatic fermentation is only 0.05%. Measurements using a monogastric animal biomimetic digestion apparatus showed that the dry matter digestibility of the fermented palm meal (duck) increased from 14.03% to approximately 35%. In vitro digestion simulation results for ruminants showed that the dry matter digestibility of the fermented palm meal prepared using this invention increased from 35.46% to approximately 75%, an increase of 111.5%.

[0131] While the specific embodiments of the present invention have been described above, they are not intended to limit the scope of protection of the present invention. Based on the technical solutions of the present invention, various modifications or variations that can be made by those skilled in the art without creative effort are still within the scope of protection of the present invention.

Claims

1. A fermented palm meal with high organic acid content, characterized in that, The palm meal contains 15wt% to 20wt% crude protein and 8wt% to 12wt% total organic acids; acid-soluble protein accounts for 25wt% to 35wt% of the crude protein content. The organic acid contains lactic acid, acetic acid, citric acid, malic acid, and succinic acid; the organic acid contains 3.9 wt% to 5 wt% lactic acid, 0.1 wt% to 1 wt% acetic acid, 2.5 wt% to 4 wt% citric acid, 0.1 wt% to 1 wt% malic acid, and 0.06 wt% to 1 wt% succinic acid; the total content of the lactic acid, acetic acid, citric acid, malic acid, and succinic acid is within 8 wt% to 12 wt%.

2. The high-organic-acid fermented palm meal according to claim 1, characterized in that, The palm meal contains 15wt% to 17wt% crude protein and 8wt% to 9wt% of various organic acids.

3. The high-organic-acid fermented palm meal according to claim 1, characterized in that, Acid-soluble protein accounts for 31 wt% to 35 wt% of crude protein.

4. The high-organic-acid fermented palm meal according to any one of claims 1-3, characterized in that, The organic acid contains 3.9 wt% to 4.5 wt% lactic acid, 0.1 wt% to 0.6 wt% acetic acid, 3 wt% to 4 wt% citric acid, 0.1 wt% to 0.8 wt% malic acid and 0.06 wt% to 0.5 wt% succinic acid.

5. The high-organic-acid fermented palm meal according to any one of claims 1-3, characterized in that, The organic acid contains 3.9 wt% to 4.3 wt% lactic acid, 0.1 wt% to 0.4 wt% acetic acid, 3.1 wt% to 3.9 wt% citric acid, 0.1 wt% to 0.5 wt% malic acid and 0.06 wt% to 0.3 wt% succinic acid.

6. A method for preparing high-organic-acid fermented palm meal according to any one of claims 1-5, characterized in that, Includes the following steps: Step 1: Preparation of seed liquid: Take 0.3-15 parts by weight of compound microbial fermentation agent and 0.4-20 parts by weight of compound enzyme preparation and mix them evenly with water to prepare seed liquid; Step 2: Inoculation and Mixing: Mix palm meal, food yeast powder, wheat bran, water and the seed solution from Step 1 evenly; Step 3: Ferment the well-mixed palm meal; and Step 4: Dry the material after fermentation to obtain fermented palm meal.

7. The preparation method according to claim 6, characterized in that, In step 1, the seed liquid contains 0.3 to 11 parts by weight of compound microbial fermentation agent and 0.4 to 16 parts by weight of compound enzyme preparation.

8. The preparation method according to claim 6, characterized in that, In step 1, the seed liquid contains 6.5 to 11 parts by weight of compound microbial fermentation agent and 7 to 12 parts by weight of compound enzyme preparation.

9. The preparation method according to claim 6, characterized in that, In step 1, the compound microbial fermentation agent is selected from one or more of yeast, Bacillus and lactic acid bacteria.

10. The preparation method according to claim 6, characterized in that, In step 1, the compound microbial fermentation agent contains yeast, Bacillus and lactic acid bacteria.

11. The preparation method according to claim 9, characterized in that, In step 1, the compound microbial fermentation agent contains 0.1 to 5 parts by weight of yeast, 0.1 to 5 parts by weight of Bacillus and 0.1 to 5 parts by weight of lactic acid bacteria.

12. The preparation method according to claim 9, characterized in that, In step 1, the compound microbial fermentation agent contains 1 to 5 parts yeast, 0.5 to 2 parts Bacillus and 4 to 5 parts lactic acid bacteria by weight.

13. The preparation method according to claim 9, characterized in that, In step 1, the yeast is selected from one or more of Saccharomyces cerevisiae, Candida utilis, and Kluyveromyces oryzae.

14. The preparation method according to claim 9, characterized in that, In step 1, the yeast contains Saccharomyces cerevisiae and Kluyveromyces oryzae.

15. The preparation method according to claim 13, characterized in that, In step 1, the yeast contains 0.05 to 4 parts by weight of Saccharomyces cerevisiae and / or 0.05 to 4 parts by weight of Kluyveromyces oryzae.

16. The preparation method according to claim 13, characterized in that, In step 1, the yeast contains 0.5 to 2.5 parts by weight of Saccharomyces cerevisiae and / or 0.5 to 2.5 parts by weight of Kluyveromyces oryzae.

17. The preparation method according to claim 9, characterized in that, In step 1, the Bacillus is selected from one or more of Bacillus subtilis, Bacillus licheniformis, Bacillus coagulans, Bacillus pumilus, and Bacillus laterosporus.

18. The preparation method according to claim 9, characterized in that, In step 1, the Bacillus species includes Bacillus pumilus and / or Bacillus coagulans.

19. The preparation method according to claim 17, characterized in that, In step 1, the Bacillus contains 0.1 to 1 part by weight of Bacillus subtilis. 0.1 to 1 part by weight of Bacillus licheniformis, 0.1 to 1 part by weight of Bacillus coagulans, 0.1 to 1 part by weight of Bacillus pumilus, and / or 0.1 to 1 part by weight of Bacillus retroflexus.

20. The preparation method according to claim 17, characterized in that, in step 1, the Bacillus contains 0.1 to 1 parts by weight of Bacillus subtilis and 0.5 to 1 part by weight of Bacillus licheniformis. 0.1 to 1 part by weight of Bacillus coagulans, 0.5 to 1 part by weight of Bacillus pumilus, and / or 0.1 to 1 part by weight of Bacillus laterosporus.

21. The preparation method according to claim 17, characterized in that, In step 1, the Bacillus used is 0.1 to 1 part by weight of Bacillus coagulans and 0.5 to 1 part by weight of Bacillus pumilus.

22. The preparation method according to claim 9, characterized in that, In step 1, the lactic acid bacteria are selected from one or more of Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus acidophilus, Enterococcus faecalis, and Enterococcus faecium.

23. The preparation method according to claim 9, characterized in that, In step 1, the lactic acid bacteria include Lactobacillus plantarum, Lactobacillus casei, Lactobacillus acidophilus, Enterococcus faecalis, and Enterococcus faecium.

24. The preparation method according to claim 22, characterized in that, In step 1, the lactic acid bacteria contain 0.5 to 1.5 parts by weight of Lactobacillus plantarum, 0.5 to 1.5 parts by weight of Lactobacillus casei, 0.5 to 1.5 parts by weight of Lactobacillus acidophilus, 0.5 to 1.5 parts by weight of Enterococcus faecalis, and / or 0.5 to 1.5 parts by weight of Enterococcus faecium.

25. The preparation method according to claim 22, characterized in that, In step 1, the lactic acid bacteria contain 1 to 1.5 parts by weight of Lactobacillus plantarum, 1 to 1.5 parts by weight of Lactobacillus casei, 0.5 to 1 part by weight of Lactobacillus acidophilus, 0.5 to 1 part by weight of Enterococcus faecalis and 0.5 to 1 part by weight of Enterococcus faecium.

26. The preparation method according to any one of claims 6-25, characterized in that, In step 1, the complex enzyme preparation is selected from one or more of protease, mannanase, cellulase and dextranase.

27. The preparation method according to any one of claims 6-25, characterized in that, In step 1, the compound enzyme preparation contains protease, mannanase, cellulase and dextranase.

28. The preparation method according to any one of claims 6-25, characterized in that, In step 1, the compound enzyme preparation contains 0.1 to 5 parts by weight of protease, 0.1 to 5 parts by weight of mannanase, 0.1 to 5 parts by weight of cellulase and 0.1 to 5 parts by weight of glucanase.

29. The preparation method according to any one of claims 6-25, characterized in that, In step 1, the compound enzyme preparation contains 4-5 parts by weight of protease, 0.5-1.5 parts by weight of mannanase, 4-5 parts by weight of cellulase and 0.1-1 parts by weight of glucanase.

30. The preparation method according to claim 26, characterized in that, In the compound enzyme preparation, in step 1, the protease activity is 50,000 to 300,000 U / g, the mannanase activity is 5,000 to 100,000 U / g, the cellulase activity is 5,000 to 100,000 U / g, and / or the dextranase activity is 5,000 to 100,000 U / g.

31. The preparation method according to any one of claims 6-25, characterized in that, In step 1, the water is 5 to 30 parts by weight.

32. The preparation method according to claim 26, characterized in that, In step 1, the water is 5 to 30 parts by weight.

33. The preparation method according to claim 28, characterized in that, In step 1, the water is 5 to 30 parts by weight.

34. The preparation method according to any one of claims 6-25, characterized in that, In step 1, the water is 5 to 15 parts by weight.

35. The preparation method according to any one of claims 6-25, characterized in that, In step 2, the amount of palm meal is 50-90 parts by weight, and the amount of food yeast powder is 10-20 parts by weight.

36. The preparation method according to claim 26, characterized in that, In step 2, the amount of palm meal is 50-90 parts by weight, and the amount of food yeast powder is 10-20 parts by weight.

37. The preparation method according to claim 28, characterized in that, In step 2, the amount of palm meal is 50-90 parts by weight, and the amount of food yeast powder is 10-20 parts by weight.

38. The preparation method according to claim 31, characterized in that, In step 2, the amount of palm meal is 50-90 parts by weight, and the amount of food yeast powder is 10-20 parts by weight.

39. The preparation method according to claim 35, characterized in that, In step 2, the amount of wheat bran is 5 to 30 parts by weight, and the amount of water is 30 to 50 parts by weight.

40. The preparation method according to any one of claims 6-25, characterized in that, In step 2, the amount of palm meal is 75-85 parts by weight, the amount of food yeast powder is 10-20 parts by weight, the amount of wheat bran is 5-10 parts by weight, and the amount of water is 35-45 parts by weight.

41. The preparation method according to any one of claims 6-25, characterized in that, In step 3, the fermentation method is anaerobic fermentation.

42. The preparation method according to claim 26, characterized in that, In step 3, the fermentation method is anaerobic fermentation.

43. The preparation method according to claim 28, characterized in that, In step 3, the fermentation method is anaerobic fermentation.

44. The preparation method according to claim 31, characterized in that, In step 3, the fermentation method is anaerobic fermentation.

45. The preparation method according to claim 35, characterized in that, In step 3, the fermentation method is anaerobic fermentation.

46. ​​The preparation method according to claim 41, characterized in that, In step 3, the anaerobic fermentation is anaerobic fermentation in a fermentation box, anaerobic fermentation in a fermentation tank, or anaerobic fermentation in a breathing bag.

47. The preparation method according to any one of claims 6-25, characterized in that, In step 3, the fermentation temperature is 25–40℃; and / or the fermentation time is 2–7 days.

48. The preparation method according to claim 41, characterized in that, In step 3, the fermentation temperature is 25–40℃; and / or the fermentation time is 2–7 days.

49. The fermented palm meal obtained by the preparation method according to any one of claims 6-48, characterized in that, The palm meal contains 15wt% to 20wt% crude protein and 8wt% to 12wt% total organic acids; of which acid-soluble protein accounts for 25wt% to 35wt% of the crude protein content.

50. A feed made from any one of claims 1-5 or claim 49, using fermented palm meal.

51. The use of fermented palm meal as described in any one of claims 1-5 or claim 49 in animal feed.

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