PROCESSO PARA FORNECER UM PRODUTO COM UM VALOR NUTRICIONAL MELHORADO A PARTIR DE UM MATERIAL RICO EM MANANO, MATERIAL RICO EM MANAMO FERMENTADO E PRODUTO PARA RAÇÃO ANIMAL

Abstract

The present invention relates to a process for providing a product having an improved nutritional value from a mannan-rich material, the method comprises the steps of (i) providing the mannan-rich material; (ii) adding at least one microorganism, in preferably at least one lactic acid bacterial strain, to the mannan-rich material, providing an inoculated mannan-rich material; and (iii) allowing the inoculated mannan-rich material to ferment for a period of at least 3 days, providing a fermented mannan-rich material having an improved nutritional value.

Description

1 / 38 PROCESS FOR PROVIDING A PRODUCT WITH NUTRITIONAL VALUE Improved from a mannan-rich material, Rich in fermented manna and a product for animal feed. TECHNICAL FIELD OF THE INVENTION

[001] The present invention relates to a process for treating a mannan-rich material (in particular, palm cake material). In particular, the present invention relates to a process for treating a mannan-rich material (in particular, palm cake material, copra cake material or guar germ cake material) to improve the nutritional value of the material. BACKGROUND OF THE INVENTION

[002] Mannan-rich materials or mannan-rich plant materials can represent an environmental problem, as the use of these mannan-rich materials is very limited and the value is very low and the nutritional value of mannan-rich materials is limited.

[003] Palm kernel cake is a material rich in mannan, with a crude protein content of about 14-18%, which is higher than that of corn and bran. Palm kernel cake comprises about 7% crude fat and 6% ash. The fibers of palm kernel cake material may comprise about 43.7% acid detergent fiber, 66.7% neutral detergent fiber, and 21.1% lignin. The fibers of palm kernel cake materials are mainly non-starch polysaccharides, and mannan is the most abundant non-starch polysaccharide, representing about 75% of the fiber content in palm kernel cake materials.

[004] Palm kernel pie can be considered Petition 870250084462, dated 09 / 19 / 2025, page 7 / 53 2 / 38 an animal feed product with a high fiber and medium-quality protein content or an animal feed supplement and, as a product or feed supplement, palm kernel cake can be classified slightly above copra cake and cocoa pod hulls due to its high protein content, but lower than fishmeal and peanut cake, because of the very low digestibility of palm kernel cake.

[005] Most agricultural industries produce large amounts of waste every year and the palm oil industry is certainly no exception, where palm kernel cake is the waste and main by-product (and waste) after the manufacture of palm oil from palm kernels.

[006] If these agricultural residues are not managed properly and effectively, they can contribute to environmental challenges and problems, as well as result in harm to human and animal health.

[007] Waste from the palm oil extraction industry constitutes a major problem, and any initiative to utilize, and perhaps even economically utilize, the waste will be a positive step towards solving environmental pollution.

[008] Many products being by-products of the agricultural industry can be used in animal feed products, and palm cake materials have been shown to provide very low nutritional value for animal feed from palm cake material, due to very low digestibility, proteins entangled in the fibrous network, presence of significant amounts of antinutritional factors (ANF).

[009] Being the main source of vegetable oil production, the oil palm (Elaeis guineensis) is one of Petition 870250084462, dated 09 / 19 / 2025, page 8 / 53 3 / 38 of the most critical trees in many tropical countries, such as Brazil, Indonesia, Malaysia, Thailand, Colombia, and Ecuador.

[0010] Studies have been conducted demonstrating that the nutritional value of palm cake materials and their effects on feed diets in the broiler industry have been evaluated, showing that the digestibility for birds is compromised due to the high fiber content and indigestion of the palm cake material, making traditional palm cake material uninteresting for non-ruminant feed.

[0011] Even for ruminants, palm cake material does not show a sufficient effect on the feed value or nutritional value when feeding diets containing palm cake material, and improvements in palm cake material would be desirable to increase the nutritional value of palm cake material, increasing digestibility, improving milk production, increasing weight gain, etc.

[0012] Thus, as the palm oil market continues to grow and annual palm kernel cake production continues to increase, more and more palm kernel cake needs to be disposed of. Furthermore, with estimated palm kernel cake production reaching 8.32 and 8.00 million tons in 2019 and 2020 worldwide, it is very urgent to find an environmentally friendly way to deal with this growing agricultural waste.

[0013] Therefore, there is a need in the industry for a process to treat mannan-rich material (in particular mannan-rich materials such as palm cake material, copra cake material or guar germ cake material) to provide a high-value product, a Petition 870250084462, dated 09 / 19 / 2025, page 9 / 53 4 / 38 product with high nutritional value, relieving the industry of a serious waste and environmental problem.

[0014] Consequently, an improved process for treating mannan-rich material (in particular mannan-rich materials such as palm cake, copra cake or guar germ cake) would be advantageous and, in particular, a more efficient and / or reliable process for treating mannan-rich materials would be advantageous, resulting in a high-value and high-nutritional-value product, relieving the industry of a serious waste problem, to the benefit of the environment. SUMMARY OF THE INVENTION

[0015] Thus, an objective of the present invention relates to a process for treating a mannan-rich material (in particular, a palm cake material, copra cake material or guar germ cake material) to improve the nutritional value of the material.

[0016] In particular, it is an objective of the present invention to provide a process for a mannan-rich material that solves the aforementioned problems of the prior art with efficiency, reliability, nutritional value, and waste and environmental issues.

[0017] Thus, one aspect of the invention relates to a process for providing a product with improved nutritional value from a mannan-rich material, wherein the method comprises the steps of:

[0018] (i) provide the mannan-rich material;

[0019] (ii) adding at least one microorganism to the mannan-rich material, providing an inoculated mannan-rich material; and Petition 870250084462, dated 09 / 19 / 2025, page 10 / 53 5 / 38

[0020] (iii) allow the inoculated material rich in mannan to ferment for a period of at least 3 days, providing a fermented mannan-rich material with improved nutritional value.

[0021] The mannan-rich material may preferably be a mannan-rich plant material or a mannan-rich algae. Preferably, the mannan-rich plant material may be palm cake material.

[0022] Another aspect of the present invention relates to a fermented mannan-rich material (in particular, a fermented palm cake material, a fermented copra cake material or a fermented guar germ cake material) comprising:

[0023] - a crude protein content in the range of 530% (w / w);

[0024] - a short-chain mannan oligomer; and

[0025] - a lactic acid concentration of at least 200 mg / 100 g of fermented palm cake material.

[0026] Yet another aspect of the present invention relates to a fermented mannan-rich material (in particular, a fermented palm cake material, a fermented copra cake material or a fermented guar germ cake material) obtained by the process according to the present invention.

[0027] Yet another aspect of the present invention relates to an animal feed product comprising a fermented mannan-rich material (in particular, a fermented palm cake material, a fermented copra cake material or a guar germ cake material). Petition 870250084462, dated 09 / 19 / 2025, p. 11 / 53 6 / 38 fermented) according to the present invention.

[0028] Another aspect of the present invention relates to the use of a fermented mannan-rich material (in particular, a fermented palm cake material, a fermented copra cake material and / or a fermented guar germ cake material) as an ingredient for an animal feed product.

[0029] The present invention will now be described in more detail below. DETAILED DESCRIPTION OF THE INVENTION

[0030] Thus, the inventors of the present invention have surprisingly found a method for providing highly nutritious products from mannan-rich materials (in particular from palm cake material, copra cake material or guar germ cake material) in an efficient and reliable process that solves the waste and environmental problems of mannan-rich materials, by subjecting the mannan-rich materials to a fermentation process using at least one microorganism, preferably at least one lactic acid bacterial strain, in combination with enzymatic treatment.

[0031] A preferred embodiment of the present invention relates to a process for providing a product with enhanced nutritional value from a mannan-rich material, wherein the method comprises the steps of:

[0032] (i) provide the mannan-rich material;

[0033] (iii) adding at least one microorganism, preferably at least one lactic acid bacterial strain, to the mannan-rich material, providing an inoculated mannan-rich material; and Petition 870250084462, dated 09 / 19 / 2025, page 12 / 53 7 / 38

[0034] (iii) allow the inoculated material rich in mannan to ferment for a period of at least 3 days, providing a fermented mannan-rich material with improved nutritional value.

[0035] Mannan-rich material can be mannan-rich plant material or mannan-rich algae.

[0036] Mannane-rich material can be polymers that contain multiple mannose sugar units as a major component. Mannane can be a polysaccharide found in hemicellulose.

[0037] Plant mannans may comprise mannose monosaccharides linked by β(1-4), occasionally with α(1-6) galactose branches, forming galactomannans. They are insoluble and a form of storage polysaccharide. Other mannan structures in plants may be a mixed mannose / glucose β(1-4) backbone. Many mannans are acetylated and some from marine sources may have sulfate ester side chains. The insoluble polysaccharides of mannans may be mannooligosaccharides.

[0038] In one embodiment of the present invention, the mannan-rich material may comprise a mannan content (in particular a β-mannan content) of at least 20 g of mannan per kg of mannan-rich material, such as at least 30 g of mannan per kg of mannan-rich material, for example, at least 40 g of mannan per kg of mannan-rich material, such as at least 50 g of mannan per kg of mannan-rich material, for example, at least 60 g of mannan per kg of mannan-rich material, such as at least 80 g of mannan per kg of mannan-rich material, for example, at least 100 g of mannan per kg of mannan-rich material, such as at least 125 g of mannan Petition 870250084462, dated 09 / 19 / 2025, page 13 / 53 8 / 38 per kg of mannan-rich material, for example, at least 150 g of mannan per kg of mannan-rich material, such as at least 175 g of mannan per kg of mannan-rich material, for example, at least 200 g of mannan per kg of mannan-rich material, such as at least 225 g of mannan per kg of mannan-rich material, for example, at least 250 g of mannan per kg of mannan-rich material, such as at least 275 g of mannan per kg of mannan-rich material, for example, at least 300 g of mannan per kg of mannan-rich material, such as at least 325 g of mannan per kg of mannan-rich material, for example, at least 350 g of mannan per kg of mannan-rich material.

[0039] Preferably, the mannan-rich material can be selected from a palm cake material, a copra cake material, a guar germ cake material, or a combination thereof.

[0040] Even more preferably, the mannan-rich material and / or the mannan-rich plant material may be palm cake material.

[0041] Palm cake material can be the residue and main byproduct after palm oil is manufactured from palm material or palm kernel material. As the palm oil market continues to expand and annual palm cake production may continue to increase, more and more palm cake material needs to be disposed of.

[0042] Preferably, the palm cake material according to the present invention may be a palm kernel cake material; a palm kernel cake; or a combination of a palm kernel cake material and a palm kernel cake. Petition 870250084462, dated 09 / 19 / 2025, page 14 / 53 9 / 38

[0043] Palm kernel oil can be the edible seed of the oil palm fruit. The fruit can produce two distinct oils: palm oil derived from the outer parts of the fruit and palm kernel oil derived from the seed of the fruit.

[0044] The pulp left after the oil has been extracted from the seed can be made into palm kernel cake. Palm kernel cake can be used as a protein-rich feed for dairy cattle or burned in boilers to generate electricity for palm oil factories and neighboring villages.

[0045] In one embodiment of the present invention, at least one microorganism can be selected from at least one lactic acid bacterial species and / or at least one yeast and / or at least one fungus. Preferably, at least one microorganism can be selected from at least one lactic acid bacterial species.

[0046] Preferably, at least one lactic acid bacterial species may be provided in step (ii) in an inoculum. The inoculum may include at least one lactic acid bacterial strain, such as at least two lactic acid bacterial strains, for example, at least 3 lactic acid bacterial strains, such as at least 4 lactic acid bacterial strains, for example, at least 5 lactic acid bacterial strains.

[0047] The inoculum according to the present invention may comprise at least 50% lactic acid bacterial strain, such as at least 60% lactic acid bacterial strain, for example, at least 70% lactic acid bacterial strain, such as at least 80% lactic acid bacterial strain, for example, at least 90% lactic acid bacterial strain Petition 870250084462, dated 09 / 19 / 2025, p. 15 / 53 10 / 38 lactic acid, such as at least 95% lactic acid bacterial strain, for example, at least 98% lactic acid bacterial strain, or 100% lactic acid bacterial strain. Preferably, the inoculum according to the present invention consists essentially of lactic acid bacterial strains.

[0048] In one embodiment of the present invention, the inoculum consists essentially of at least one lactic acid bacterial strain.

[0049] In a preferred embodiment of the present invention, at least one lactic acid bacterial strain may be selected from the group consisting of the genera Enterococcus, Lactobacillus, Pediococcus, Lactococcus or Bifidobacterium or combinations thereof.

[0050] In a further embodiment of the present invention, at least one lactic acid bacterial strain may be selected from the group consisting of Pediococcus pentosaceus; Pendiococcus acidilactici; Lactobacillus plantarum, Lactobacillus rhamnosus, Enterococcus faecium, Lactobacillus acidophilus, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus salivarius, Lactobacillus pentosus, Lactobacillus vaginalis, Lactobacillus xylosus and a combination thereof.

[0051] In one embodiment of the present invention, at least one lactic acid bacterial strain may be the main microorganism present in the fermented palm cake material and / or in the animal feed product according to the present invention. Even more preferably, the main lactic acid bacterial strains may be selected from the groups consisting of Pediococcus pentosaceus; Pediococcus acidilactici, Lactobacillus Petition 870250084462, dated 19 / 09 / 2025, p. 16 / 53 11 / 38 plantarum, Lactobacillus rhamnosus, Enterococcus faecium, Lactobacillus acidophilus, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium bifidum, Lactobacillus salivarius, Lactobacillus pentoses, Lactobacillus vaginalis and Lactobacillus xylosus.

[0052] At least one lactic acid bacterial strain may be selected from the group consisting of one or more of the following: Pediococcus pentosaceus (DSM 12834); Pendiococcus acidilactici (DSM 16243); Lactobacillus plantarum (DSM 12837); Enterococcus faecium (NCIMB 30122), Lactobacillus rhamnosus (NCIMB 30121), Pediococcus pentosaceus HTS (LMG P22549), Pendiococcus acidilactici (NCIMB 30086) and / or Lactobacillus plantarum LSI (NCIMB 30083). Preferably, one or more lactic acid bacterial strains may be selected from the group consisting of one or more of the following: Pediococcus pentosaceus (DSM 12834); Pendiococcus acidilactici (DSM 16243); Lactobacillus plantarum (DSM 12837).

[0053] Preferably, at least one lactic acid bacterial strain and / or the main microorganism (as provided in step (ii) and / or as provided in the fermented mannan-rich material (in particular, a fermented palm cake material, a fermented copra cake material or a fermented guar germ cake material) and / or as provided in the animal feed product according to the present invention may include one or more strains of Lactobacillus plantarum.

[0054] In the context of the present invention, the term principal microorganism refers to the microorganism present in the greatest quantity, determined in a weight / weight ratio relative to the total number of microorganisms present, by Petition 870250084462, dated 09 / 19 / 2025, page 17 / 53 12 / 38 example, during co-fermentation.

[0055] Co-fermentation can be a mixture of different microorganisms (such as a mixture of yeasts, fungi and / or bacteria) or a mixture of different bacteria.

[0056] Fermentation can be a co-fermentation comprising a combination of microorganisms selected from:

[0057] - at least one lactic acid bacterial species and at least one yeast; or

[0058] - at least one lactic acid bacterial species and / or at least one fungus; or

[0059] - at least one yeast and at least one fungus; or

[0060] - at least one lactic acid bacterial species, at least one yeast and at least one fungus.

[0061] The yeast may be selected from Saccharomyces spp., in particular a strain of Saccharomyces cerevisiae or Saccharomyces uvarum; a strain of Pichia, preferably Pichia stipitis; a strain of Candida, in particular a strain of Candida utilis, Candida diddensii or Candida boidinii; a strain of Zymomonas; a strain of Hansenula, in particular H. anomala; a strain of Klyveromyces, in particular K. fragilis; or a strain of Schizosaccharomyces, in particular S. pombe. Preferably, the yeast may be selected from Saccharomyces spp., especially from a strain of Saccharomyces cerevisiae.

[0062] The fungus can be selected from Aspergillus.

[0063] In one embodiment of the present invention, the inoculum, the animal feed product and / or the material rich in Petition 870250084462, dated 09 / 19 / 2025, page 18 / 53 13 / 38 fermented mannan (in particular, fermented palm cake material, fermented copra cake material or fermented guar germ cake material) does not comprise a yeast.

[0064] Preferably, the fermentation process according to the present invention (i.e., step iii) does not include yeast fermentation. Preferably, the fermentation process according to the present invention (i.e., step iii) does not include fungal fermentation. Preferably, the fermentation process according to the present invention (i.e., step iii) does not include yeast fermentation and does not include fungal fermentation.

[0065] The fermentation process according to the present invention (step iii) may preferably be essentially a homofermentative process. Essentially homofermentative means that the predominant bacterial flora driving the fermentation is homofermentative to produce mainly a single organic acid. Preferably, the single organic acid may be lactic acid.

[0066] In the present context, the term essentially homofermentative refers to a fermentation process in which 60% or more of the bacterial strains are homofermentative, such as 70% or more of the bacterial strains are homofermentative, for example, 80% or more of the bacterial strains are homofermentative, such as 85% or more of the bacterial strains are homofermentative, for example, 90% or more of the bacterial strains are homofermentative, such as 95% or more of the bacterial strains are homofermentative, for example, 99% or more of the bacterial strains are homofermentative.

[0067] In one embodiment of the present invention, the Petition 870250084462, dated 09 / 19 / 2025, p. 19 / 53 14 / 38 The fermentation process provided in step (iii) may be a homolactic fermentation. Homolactic fermentation may result in one molecule of glucose being ultimately converted into two molecules of lactic acid. Heterolactic fermentation, in contrast, produces carbon dioxide, ethanol, and acetic acid, in addition to lactic acid. Preferably, the level of heterolactic fermentation may be limited and homolactic fermentation may be favored.

[0068] The term homolactic fermentation, when used in accordance with the present invention, indicates that the main product of fermentation, the main organic acid produced, is lactic acid, and the levels of acetic acid and ethanol are below the taste threshold, around the taste threshold, or slightly above the taste threshold.

[0069] Preferably, essentially homofermentative may indicate a ratio of lactic acid to acetic acid or lactic acid to ethanol in (mM / mM) of more than 1:1, such as 2:1 or more, for example, 10:1 or more, such as 20:1 or more, for example, 50:1 or more, or such as 100:1 or more.

[0070] It may be preferable that the formation of acetic acid and / or ethanol during fermentation be reduced or omitted, since these products are considered to negatively affect the flavor of the resulting fermented mannan-rich material (in particular, fermented palm cake material, fermented copra cake material or fermented guar germ cake material) and / or the animal feed product comprising the fermented mannan-rich material (in particular, fermented palm cake material, fermented copra cake material or fermented guar germ cake material) in accordance with Petition 870250084462, dated 09 / 19 / 2025, p. 20 / 53 15 / 38 the present invention.

[0071] Therefore, to reduce or avoid the formation of acetic acid and / or ethanol during the fermentation of mannan-rich material, as provided in step (i) of the present invention, oxygen, in particular gaseous oxygen, can be removed (totally or partially) from the fermenting material.

[0072] The process of removing or depleting oxygen from the inoculated material rich in mannan can be carried out during at least the first hour of fermentation; such as within at least the first 5 days of fermentation; for example, within at least the first 2 days of fermentation; such as within at least the first 24 hours of fermentation; for example, within at least the first 18 hours of fermentation; such as within at least the first 12 hours of fermentation; for example, within at least the first 6 hours of fermentation; such as within at least the first 3 hours of fermentation; for example, within at least the first 2 hours of fermentation; such as within at least the first hour of fermentation; for example, throughout the entire fermentation process.

[0073] In one embodiment of the present invention, the amount of oxygen present in and / or around the inoculated mannan-rich material can be reduced in, or depleted of, oxygen by the introduction of gas depletion and / or vacuum. In the context of the present invention, the term gas depletion refers to a process in which a gas other than oxygen can be introduced into the inoculated material and pushes and depletes (totally or partially) the oxygen from the inoculated mannan-rich material. Alternatively, or additionally, the rich material Petition 870250084462, dated 09 / 19 / 2025, page 21 / 53 16 / 38 in inoculated mannan may have its oxygen content reduced or depleted by subjecting the inoculated mannan-rich material to vacuum treatment.

[0074] The fermentation process of an oxygen-depleted mannan-rich inoculated material may result in a fermented mannan-rich material with a reduced or no detectable amount of acetic acid and / or ethanol.

[0075] In palm kernel cake material, particularly palm kernel cake, the cell wall may comprise 55-65% (about 58%) mannan, 8-15% (in particular about 12%) cellulose and 1-8% (in particular about 4%) xylan. The mannan structure of palm kernel cake material, particularly palm kernel cake material, may be hard and crystalline.

[0076] In one embodiment of the present invention, at least one enzyme can be added to the mannan-rich material (in particular, palm cake material, copra cake material or guar germ cake material) in combination with at least one lactic acid bacterial strain in step (ii).

[0077] At least one enzyme may include at least one cell wall-degrading enzyme.

[0078] The enzyme that degrades the cell wall may be an enzyme capable of cleaving the cell wall polysaccharides into smaller units or shorter chains.

[0079] The number of carbohydrate residues in a carbohydrate chain can be expressed as the degree of polymerization (DP). This means that an oligosaccharide composed of two mannose residues has a degree of polymerization (DP) of 2. Petition 870250084462, dated 09 / 19 / 2025, page 22 / 53 17 / 38

[0080] In one embodiment of the present invention, at least one enzyme added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii) may be β-mannanase (or β-mannosidase or β-D-mannoside mannohydrolase).

[0081] Preferably, at least one enzyme added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii) may be selected from the group consisting of glucanase (e.g., endo-1,3(4)-e-glucanase), xylanase (e.g., endo-1,4-xylanase), cellulase, phosphatase (e.g., 6-phytase) and a combination thereof.

[0082] In one embodiment of the present invention, at least one enzyme added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii) may be a combination of glucanase (e.g., endo-1,3(4)-e-glucanase), xylanase (e.g., endo-1,4-xylanase), cellulase and phosphatase (e.g., 6-phytase).

[0083] At least one enzyme added to the mannan-rich material, in combination with at least one lactic acid bacterial strain in step (ii), may preferably be a combination of β-mannanase, glucanase (e.g., endo-1,3(4)-β-glucanase), xylanase (e.g., endo-1,4-xylanase), cellulase and phosphatase (e.g., 6-phytase).

[0084] In one embodiment of the present invention, at least one enzyme added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii) may be a mixture of enzymes, such as a series Petition 870250084462, dated 09 / 19 / 2025, page 23 / 53 18 / 38 Ronozyme is supplied by DSM.

[0085] In the present context, the term fermentation or fermented refers to a controlled metabolic process of mannan-rich material to be fermented by adding a predetermined amount of inoculum to the mannan-rich material, allowing microorganisms, in particular at least one lactic acid bacterial strain, and the mannan-rich material to interact, resulting in the degradation of the mannan-rich material.

[0086] In one embodiment of the present invention, the fermentation is a solid-state fermentation.

[0087] The inventors of the present invention have surprisingly discovered that the inoculated mannan-rich material should preferably be fermented for an extended period, of more than 3 days, in order to provide sufficient decomposition of the mannan-rich material and to provide the product with enhanced nutritional value.

[0088] The mannan-rich material inoculated according to the present invention can be left to ferment for more than 3 days, such as about 4 days or more, for example, for about 5 days or more, such as about 6 days or more, for example, for about 8 days or more, such as about 10 days or more, for example, for about 12 days or more, such as about 14 days or more, for example, for about 16 days or more, such as about 18 days or more, for example, for about 20 days or more, such as about 22 days or more, for example, for about 24 days or more, such as about 26 days or more, for example, for about 28 days or more, such as about 30 days or more, for example, for about 32 days or more, such as about 35 days or more, for example, Petition 870250084462, dated 09 / 19 / 2025, p. 24 / 53 19 / 38 for about 37 days or more, such as for about 40 days or more, for example, for about 42 days or more, such as for about 45 days or more, for example, in the 5-45 day range, such as in the 10-40 day range, for example, in the range of 20-35 days, as in the 25-30 day interval.

[0089] In one embodiment of the present invention, a second portion of at least one lactic acid bacterial strain can be added to the inoculated mannan-rich material at certain intervals, to provide or maintain a high fermentative effect.

[0090] The correct intervals can be determined by the time elapsed since the last addition of at least one lactic acid bacterial strain and / or by determining a drop in activity or fermentation effect.

[0091] In one embodiment of the present invention, the addition of the second portion of at least one lactic acid bacterial strain can be carried out from 1 to 10 times during the entire fermentation period provided in step (iii), such as 2-8 times, for example, 3-6 times, or 4-5 times.

[0092] In one embodiment of the present invention, the time period of the combined fermentation process and enzymatic treatment provided in step (iii) may be longer than 3 days, such as about 4 days or more, for example, about 5 days or more, such as about 6 days or more, for example, about 8 days or more, such as about 10 days or more, for example, about 12 days or more, such as about 14 days or more, for example, about 16 days or more, such as about 18 days or more, for example, about 20 days or more, such as about 22 days or Petition 870250084462, dated 09 / 19 / 2025, p. 25 / 53 20 / 38 more, for example, for about 24 days or more, such as for about 26 days or more, for example, for about 28 days or more, such as for about 30 days or more, for example, for about 32 days or more, such as for about 35 days or more, for example, for about 37 days or more, such as for about 40 days or more, for example, for about 42 days or more, such as for about 45 days or more, for example, in the interval of 5 to 45 days, such as in the interval of 10 to 40 days, for example, in the interval of 20 to 35 days, such as in the interval of 25 to 30 days.

[0093] The inoculated mannan-rich material (in particular, inoculated palm cake material, inoculated copra cake material or inoculated guar germ cake material) can be fermented at a temperature in the range of 15-45 °C, such as 15-40 °C, such as 25-35 °C, such as 30-40 °C, such as 15-20 °C or such as 40-45 °C.

[0094] The combined fermentation process and enzymatic treatment provided for in step (iii) may be carried out at a temperature in the range of 15-45 °C, such as 15-40 °C, such as 25-35 °C, such as 30-40 °C, such as 15-20 °C or such as 40-45 °C.

[0095] In one embodiment of the present invention, the moisture content of the inoculated mannan-rich material can be adjusted, before it is allowed to ferment (step (iii)), to a moisture content of the inoculated mannan-rich material in the range of 15-70% (w / w) moisture, for example, in the range of 20-65% (w / w) moisture; as in the range of 30-55% (w / w) moisture; for example, in the range of 35-45% (w / w) moisture; as about 40% (w / w) moisture.

[0096] The moisture content of the inoculated mannan-rich material provided in step (ii) and / or during the Petition 870250084462, dated 09 / 19 / 2025, page 26 / 53 21 / 38 fermentation process in step (iii) may be in the range of 15-70% (w / w) moisture, for example, in the range of 20-65% (w / w) moisture; as in the range of 30-55% (w / w) moisture; For example, in the range of 35-45% (w / w) moisture; such as around 40% (w / w) moisture.

[0097] In one embodiment of the present invention, the inoculated mannan-rich material can be allowed to ferment until the pH of the fermenting composition has reached a pH value below 6.5; such as a pH value below 6.0; for example, a pH value below 5.5; such as a pH value below 5.0; for example, a pH value below 4.5; such as a pH value below 4.3; for example, a pH value below 4.1; such as a pH value below 4.0; for example, a pH value below 3.9; such as a pH value below 3.8.

[0098] In another embodiment of the present invention, the fermented mannan-rich material may have a pH value below 6.5; such as a pH value below 6.0; for example, a pH value below 5.5; such as a pH value below 5.0; for example, a pH value below 4.5; such as a pH value below 4.3; for example, a pH value below 4.1; such as a pH value below 4.0; for example, a pH value below 3.9; such as a pH value below 3.8.

[0099] In one embodiment of the present invention, the mannan-rich material provided in step (i) can be ground into smaller particle sizes. Preferably, several particle fractions can be provided, for example, by sieving.

[00100] Preferably, most of the ground mannan-rich material can have a particle size below Petition 870250084462, dated 09 / 19 / 2025, page 27 / 53 22 / 38 of 1.0 mm, such as below 0.9 mm, for example, below 0.8 mm.

[00101] In one embodiment of the present invention, the ground mannan-rich material comprises in the range of 30-70% (w / w); as in the range of 40-60% (w / w); for example, about % (w / w) of the composition comprises a particle size below 0.5 mm and in the range of 30-70% (w / w); as in the range of 40-60% (w / w); for example, about 50% (w / w) of the ground mannan-rich material comprises a particle size above 0.5 mm.

[00102] Preferably, in the range of 20-80% (w / w) of the ground mannan-rich material, it comprises a particle size below 0.5 mm (preferably in the range of 0.0-10.5 mm), and in the range of 20-80% (w / w) of the ground mannan-rich material according to the present invention, it comprises a particle size above 0.5 mm (preferably in the range of 0.5-1.0 mm)

[00103] In a further embodiment of the present invention, the ground mannan-rich material according to the present invention comprises at least 2, preferably at least 3, even more preferably at least 4 of the following criteria:

[00104] a) 1-10 % (m / m), as about 5 % (m / m), of the ground mannan-rich material comprises a particle size above 1.0 mm;

[00105] b) 45-55% (w / w), as about 50% (w / w), of the ground mannan-rich material comprises a particle size between 0.5-1.0 mm;

[00106] c) 30-40% (w / w), as approximately 50% (w / w) of the ground mannan-rich material comprises a size of Petition 870250084462, dated 09 / 19 / 2025, page 28 / 53 23 / 38 particle between 0.25-0.5 mm; and / or

[00107] d) 5-15 % (w / w), as approximately 10 % (w / w) of the ground mannan-rich material comprises a particle size below 0.25 mm.

[00108] In the present context, the term "over" refers to a variation from the stated value of 10% or less, such as 5% or less, for example, 1% or less.

[00109] The selection of the various particle sizes can be determined by sieving, as is known to the skilled person.

[00110] In one embodiment of the present invention, the fermented mannan-rich material can be separated into at least 2 fractions, as well as at least 3 fractions, for example, at least 4 fractions.

[00111] A preferred embodiment of the present invention relates to a fermented mannan-rich material obtained by the process according to the present invention.

[00112] A preferred additional embodiment of the present invention relates to a fermented mannan-rich material (in particular, a fermented palm cake material, a fermented copra cake material, a fermented guar germ cake material or a combination thereof) comprising:

[00113] - a crude protein content in the range of 530% (w / w);

[00114] - a short-chain mannan oligomer; and

[00115] - a lactic acid concentration of at least 200 mg / 100 g of fermented palm cake material.

[00116] In one embodiment of the present invention, the Petition 870250084462, dated 09 / 19 / 2025, p. 29 / 53 24 / 38 fermented mannan-rich material (or animal feed product) may additionally comprise at least one lactic acid bacterium or a derivative thereof.

[00117] At least one lactic acid bacterium or a derivative thereof may include viable cells of at least one lactic acid bacterium, dead or inactivated cells of at least one lactic acid bacterium, and / or a fraction obtained from at least one lactic acid bacterium. Preferably, the lactic acid bacterium derivative, in the context of the present invention, does not include lactic acid and / or acetic acid, as these components are defined elsewhere in this application.

[00118] Fermented mannan-rich material (in particular, fermented palm cake material, fermented copra cake material or fermented guar germ cake material, preferably fermented palm cake material) may contain a crude protein content in the range of 5-30% (w / w) of the fermented mannan-rich material, such as in the range of 7-25% (w / w), for example, in the range of 10-20% (w / w), such as in the range of 12-18% (w / w), for example, in the range of 14-17% (w / w), such as about 16% (w / w).

[00119] Fermented mannan-rich material (in particular, fermented palm cake material, fermented copra cake material or fermented guar germ cake material, preferably fermented palm cake material) may contain a crude fiber content in the range of 530% (w / w) of the fermented mannan-rich material, such as in the range of 7-25% (w / w), for example, in the range of 10-20% (w / w), such as in the range of 12-18% (w / w), for example, in the range of 14-17% (w / w), such as about 16% (w / w).

[00120] Fermented mannan-rich material (in Petition 870250084462, dated 09 / 19 / 2025, page 30 / 53 25 / 38 in particular, the fermented palm cake material, the fermented copra cake material or the fermented guar germ cake material, preferably the fermented palm cake material) may contain a crude fat content in the range of 1-15% (w / w) of the fermented mannan-rich material, such as in the range of 2-12% (w / w), for example, in the range of 3-10% (w / w), such as in the range of 4-8% (w / w), for example, in the range of 5-7% (w / w), such as about 6% (w / w).

[00121] In one embodiment of the present invention, the short-chain mannan oligomer may have a degree of polymerization (DP) in the range of 2-10, such as in the range of 2-8, for example, in the range of 2-6, such as in the range of 2-4.

[00122] The mannose subunits present in the mannan-rich fermentation material may preferably be present as short-chain mannan oligomer. Preferably, at least 10% of the mannose subunits present in the mannan-rich fermentation material may be present as short-chain mannan oligomer, such as at least 20%, for example, at least 30%, such as at least %, for example, at least 50%, such as at least 60%, for example, at least 65%, such as at least 70%, for example, at least 75%, such as at least 80%, for example, at least %, such as at least 90%, for example, at least 95%. The short-chain mannan oligomer may preferably have a degree of polymerization (DP) in the range of 2 to 10, such as in the range of 2 to 8, for example, in the range of 2 to 6, or in the range of 2 to 4.

[00123] The animal feed product and / or the mannan-rich fermentation material may have a high content of lactic acid bacterial strains, preferably a high content Petition 870250084462, dated 09 / 19 / 2025, page 31 / 53 26 / 38 of viable lactic acid bacterial strain.

[00124] In one embodiment of the present invention, the animal feed product and / or the fermented mannan-rich material comprises at least one lactic acid bacterial strain in a total amount in the range of 105-1012 CFU per gram, such as in the range of 106-1012 CFU per gram, for example, in the range of 107-1011 CFU per gram, such as in the range of 108-1011 CFU per gram, for example, in the range of 109-1010 CFU per gram.

[00125] The fermented palm cake material and / or animal feed product according to the present invention may comprise a lactic acid concentration of at least 50 mM, as at least 100 mM, as at least 250 mM, as at least 500 mM, as at least 750 mM, as at least 1000 mM, as in the range of 100-1000 mM, as in the range of 100-500 mM, as in the range of 100-300 mM, as in the range of 100-200 mM, as in the range of 150-500 mM, as in the range of 200-500 mM or as in the range of 300-500 mM of lactic acid.

[00126] In one embodiment of the present invention, the fermented palm cake material and / or the animal feed product according to the present invention may contain a lactic acid concentration of at least 200 mg / 100 g of fermented palm cake material (or animal feed product), such as at least 400 mg / 100 g, for example, at least 600 mg / 100 g, such as at least 800 mg / 100 g, for example, at least 1,000 mg / 100 g, such as at least 1,500 mg / 100 g, for example, at least 2,000 mg / 100 g, such as at least 2,500 mg / 100 g, for example, at least 3,000 mg / 100 g, such as at least 3,500 mg / 100 g, for example, at least 4,000 mg / 100 g, such as at least 4,500 mg / 100 g, for example, at least 5,000 mg / 100 g, as at least 5,500 mg / 100 g, for example, by Petition 870250084462, dated 09 / 19 / 2025, page 32 / 53 27 / 38 less than 6,000 mg / 100 g, such as at least 6,500 mg / 100 g, for example, at least 7,000 mg / 100 g, such as at least 7,500 mg / 100 g, for example, at least 8,000 mg / 100 g, such as at least 8,500 mg / 100 g, for example, at least 9,000 mg / 100 g, such as in the range of 200-9,000 mg / 100 g, for example, in the range of 1,000-8,500 mg / 100 g, such as in the range of 2,000-8,000 mg / 100 g, for example, in the range of 2,500-7,500 mg / 100 g, such as in the range of 3,000-7,000 mg / 100 g, for example, in the range of 3,500-6,500 mg / 100g, as in the range of 4,500-6,000 mg / 100g, for example, in the range of 5,000-5,500 mg / 100g.

[00127] In one embodiment of the present invention, the fermented mannan-rich material may have an acetic acid and / or ethanol content in the range of 0-1% (w / w) of acetic acid or ethanol relative to the fermented mannan-rich material, such as in the range of 0.1-0.75% of acetic acid or ethanol relative to the fermented mannan-rich material, such as in the range of 0.25-0.5% of acetic acid or ethanol relative to the fermented mannan-rich material, such as in the range of 0.0001-0.001% of acetic acid or ethanol relative to the fermented mannan-rich material, such as in the range of 0.0005-0.05% of acetic acid or ethanol relative to the fermented mannan-rich material. Preferably, the fermented ingredient may have an undetectable content of acetic acid or ethanol.

[00128] In another embodiment of the present invention, the fermented mannan-rich material may have an acetic acid or ethanol content of up to 100 mM; as up to 50 mM; for example, up to 25 mM; as up to 10 mM; for example, up to 1 mM; as up to 0.1 mM; for example, up to 0.01 mM; as in the range of 0.01-100 mM; as in the range of 0.1-75 mM; as in the range of 1-50 mM; as in the range Petition 870250084462, dated 09 / 19 / 2025, page 33 / 53 28 / 38 from 5-25 mM; as in the 1-10 mM range.

[00129] Due to their moderate crude protein content, mannan-rich materials may be suitable for animal feed products and may totally or partially replace soybean meal and other protein-rich materials, for example, in the breeding industry. Mannan-rich materials can be widely used as a feed supplement.

[00130] Untreated mannan-rich material may have a digestibility of less than 30%, and the traditional challenge of using mannan-rich materials may be that, once the mannan from the mannan-rich material enters the intestines of animals, it is very difficult to break down for utilization and energy supply, and may even cause negative effects on animal health.

[00131] The high fiber content in mannan-rich materials (in particular, palm cake, copra cake or guar germ cake, preferably palm cake) can be closely bound to and enveloped by proteins, which strongly affects the digestibility and absorption of proteins in animals during consumption. Therefore, mannan-rich materials used in animal feed have been difficult for animals to digest and absorb.

[00132] Consequently, the nutritional value of materials rich in mannan can be greatly improved by destroying their fiber structure and degrading their mannan.

[00133] The degradation of mannan-rich materials and the destruction of fiber structure can result in increased solubility and, consequently, greater digestibility of mannan-rich materials. Petition 870250084462, dated 09 / 19 / 2025, page 34 / 53 29 / 38

[00134] Preferably, the solubility and digestibility of materials rich in mannan and / or mannan-oligosaccharides can, after being subjected to fermentation according to the present invention, be increased by at least 20%, such as at least 25%, for example, at least 30%, such as at least 35%, for example, at least 40%, such as at least 45%, for example, at least 50%, such as at least 55%, for example, at least 60%, such as at least 70%, for example, in the range of 20-70%, such as in the range of 25-65%, for example, in the range of 30-60%, such as in the range of 35-55%, for example, in the range of 40-50%, such as about 48%.

[00135] The solubility and digestibility of materials rich in mannan and / or mannan-oligosaccharides can be increased from about 15% solubility to a solubility, after being subjected to fermentation according to the present invention, of at least 20%, such as at least 25%, for example, at least 30%, such as at least 35%, for example, at least 40%, such as at least 45%, for example, at least 50%, such as at least 55%, for example, at least 60%, such as at least 70%, for example, in the range of 20-70%, such as in the range of 25-65%, for example, in the range of 30-60%, such as in the range of 35-55%, for example, in the range of 40-50%, such as about 48%.

[00136] In one embodiment of the present invention, the amount of soluble material rich in mannan and / or mannan-oligosaccharides can be at least 25% greater after being subjected to fermentation according to the present invention compared to the material rich in mannan and / or mannan-oligosaccharides not subjected to fermentation according to the present invention, such as at least 50% greater, for example, at least 75% greater, Petition 870250084462, dated 09 / 19 / 2025, page 35 / 53 30 / 38 as at least 100% larger, for example, at least 150% larger, as at least 200% larger, for example, at least 250% larger, as at least 300% larger, for example, at least 350% larger, as at least 400% larger, for example, at least 450% larger.

[00137] In one embodiment of the present invention, solubility can be determined and / or quantified by high-performance liquid chromatography (HPLC).

[00138] In one embodiment of the present invention, the mannan-rich fermentation material may exhibit a pepsin digestibility of at least 50%, such as at least 60%, for example, at least 65%, such as at least 70%, for example, at least 75%, such as at least 80%, for example, at least 85%, such as at least 90%, for example, at least 92%.

[00139] However, in one embodiment of the present invention, the fermented mannan-rich material may have an ileal digestibility of at least 50%, such as at least 60%, for example, at least 65%, such as at least 70%, for example, at least 75%, such as at least 80%, for example, at least 85%, such as at least 90%, for example, at least 92%.

[00140] Still, in one embodiment of the present invention, the fermented mannan-rich material may have an in vivo digestibility—determined in a cow—of at least 50%, such as at least 60%, for example, at least 65%, such as at least 70%, for example, at least 75%, such as at least 80%, for example, at least 85%.

[00141] In one embodiment of the present invention, the digestibility of the fermented mannan-rich material can be Petition 870250084462, dated 09 / 19 / 2025, pp. 36 / 53 31 / 38 at least 10% greater than the digestibility of the mannan-rich material provided in step (i), being at least 20% greater, for example, at least 30% greater, being at least 40% greater, for example, at least 50% greater, being at least 60% greater, for example, at least 70% greater, being at least 80% greater, for example, at least 90% greater, being at least 100% greater, for example, at least 110% greater, being at least 120% greater, for example, at least 130% greater.

[00142] Digestibility can be determined in vivo in an animal, such as a cow or a pig, or digestibility can be determined by an in vitro assay.

[00143] Pepsin digestibility can be measured according to ISO 6655 (August 1997) using, as per said standard, a pepsin concentration of 0.02% in hydrochloric acid.

[00144] Ileal digestibility (also referred to as Boisen digestibility) can be measured according to the methods described by S. Boisen in Prediction of the apparent ileal digestibility of protein and amino acid in feedstuffs for pigs by in vitro analysis, Animal Feed Science Technology, 51, pp. 29-43 (1995) and subsequently described in In vitro analysis for determining standardized ileal digestibility of protein and amino acids in actual batches of feedstuffs and diets for pigs; Ciência do Pecuária, 309: pp. 182-185 (2007).

[00145] Fermented mannan-rich material (in particular, fermented palm cake material, fermented copra cake material or fermented guar germ cake material, preferably palm cake material) Petition 870250084462, dated 09 / 19 / 2025, pp. 37 / 53 32 / 38 fermented) according to the present invention may comprise in the range of 30-70% (w / w); as well as in the range of 40-60% (w / w); For example, approximately 50% (w / w) of the mannan-rich fermented material with a particle size of less than 0.5 mm and a range of 30-70% (w / w); such as a range of 40-60% (w / w); for example, approximately 50% (w / w) of the mannan-rich fermented material with a particle size greater than 0.5 mm.

[00146] Preferably, in the range of 20-80% (w / w) of the mannan-rich fermentation material according to the present invention, it comprises a particle size below 0.5 mm (preferably in the range of 0.01-0.5 mm), and in which in the range of 20-80% (w / w) of the fermented mannan-rich material according to the present invention comprises a particle size above 0.5 mm (preferably in the range of 0.5-1.0 mm).

[00147] In a further more advanced embodiment of the present invention, the fermented mannan-rich material according to the present invention comprises at least 2, preferably at least 3, even more preferably at least 4 of the following criteria:

[00148] a) 1-10 % (m / m), as approximately 5 % (m / m) of the mannan-rich fermentation material comprises a particle size above 1.0 mm;

[00149] b) 45-55 % (m / m), as approximately 50 % (m / m) of the fermented mannan-rich material comprises a particle size between 0.5-1.0 mm;

[00150] c) 30-40% (w / w), as approximately 50% (w / w) of the fermented mannan-rich material comprises a particle size between 0.25-0.5 mm; and / or

[00151] d) 5-15% (w / w), as approximately 10% (w / w) of the fermented mannan-rich material comprises a size Petition 870250084462, dated 09 / 19 / 2025, pp. 38 / 53 33 / 38 of particles smaller than 0.25 mm.

[00152] In the present context, the term "over" refers to a variation from the stated value of 10% or less, such as 5% or less, for example, 1% or less.

[00153] The selection of the various particle sizes can be determined by sieving, as is known to the skilled person.

[00154] In another embodiment of the present invention, the fermented mannan-rich material can be dried, providing a dry fermented mannan-rich material.

[00155] Fermented mannan-rich dry material may include a moisture content in the range of 1-10% (w / w), such as in the range of 2-9% (w / w), for example, in the range of 3-8% (w / w), such as in the range of 4-7% (w / w), for example, in the range of 4.5-6% (w / w), such as about 5% (w / w) moisture.

[00156] Drying of fermented mannan-rich material can be carried out using a gentle drying method, such as centrifugal flash drying, spray drying, drum drying, ring drying or vacuum drying to protect the nutritional value of the material.

[00157] A preferred embodiment of the present invention relates to an animal feed product comprising a fermented mannan-rich material according to the present invention.

[00158] The animal feed product according to the present invention may comprise in the range of 5-95% (w / w) of the fermented mannan-rich material according to the present invention, as in the range of 10-90% (w / w), for example, in the range of 15-85% (w / w), as in the range of 20-80% (w / w), for example, in the range of 25-75% (w / w), as in the range of 30-70% (w / w), for example Petition 870250084462, dated 09 / 19 / 2025, pp. 39 / 53 34 / 38 example, in the range of 35-65% (p / p), as in the range of 40-60% (p / p), for example, in the range of 45-55% (p / p), as approximately % (p / p).

[00159] In one embodiment of the present invention, the animal feed product may include a carbohydrate component. Preferably, the carbohydrate component may include molasses.

[00160] The animal feed product according to the present invention may comprise in the range of 0.5-10% (w / w) of carbohydrate component, such as in the range of 1-9% (w / w), for example, in the range of 2-8% (w / w), such as in the range of 3-7% (w / w), for example, in the range of 4-6% (w / w), such as about 5% (w / w).

[00161] In one embodiment of the present invention, the animal feed product may include grass, hay, or a combination of grass and hay.

[00162] The animal feed product according to the present invention may comprise in the range of 5-90% (w / w) of grass, hay or a combination of grass and hay, such as in the range of 10-80% (w / w), for example, in the range of 20-70% (w / w), such as in the range of 30-60% (w / w), for example, in the range of 35-50% (w / w), such as in the range of 40-45% (w / w), for example, about % (w / w).

[00163] In one embodiment of the present invention, the animal feed product may contain limestone, a mineral premix, a salt, trace elements, a vitamin premix, or a mixture thereof.

[00164] In one embodiment of the present invention, the digestibility of fermented mannan-rich material (in particular, fermented palm cake material, the material Petition 870250084462, dated 09 / 19 / 2025, pp. 40-53 35 / 38 of fermented copra cake or fermented guar germ cake material, preferably fermented palm cake material) can be 50% or more, such as 55% or more, for example, 60% or more, such as 65% or more, for example, 70% or more, such as 75% or more, for example, 80% or more.

[00165] The animal feed product according to the previous invention may be a farm animal feed product or a pet food product.

[00166] Preferably, the livestock feed product may be a pig feed product (such as a sow feed product and / or a piglet feed product), a broiler feed product, a chicken feed product, a rooster feed product, a laying hen feed product, a turkey feed product, a duck feed product, a goose feed product, a quail feed product, a ruminant feed product (such as a cattle feed product, a sheep feed product, a goat feed product), a horse feed product, a fish feed product.

[00167] In the context of the present invention, the term "comprising," which may be synonymous with the terms "including," "containing," or "characterized by," refers to an inclusive or open list of features and does not exclude additional unrecorded features or steps of the method. The term "understanding" leaves the claim open to the inclusion of unspecified ingredients, even in large quantities.

[00168] In the context of the present invention, the terms consisting essentially of and consists of Petition 870250084462, dated 09 / 19 / 2025, pp. 41 / 53 36 / 38 essentially refer to limiting the scope of a claim to the features or steps specified, and to those features or steps not mentioned that do not materially affect the basic and innovative characteristics of the claimed invention. EXAMPLES

[00169] Example 1 - Influence of fermentation and fermentation time on the solubility of mannan-rich materials obtained from palm kernel cake.

[00170] For this experiment, palm kernel cake was obtained as a byproduct of palm kernel oil production.

[00171] Two conventional preparations were made (Sample 1 and Sample 2). The first (sample 1) comprises 60 kg of palm kernel cake and 40 kg of water. The second (sample 2) comprises 60 kg of palm kernel cake and 40 kg of water mixed with the enzyme β-mannanase.

[00172] A preparation (Sample 3) according to the present invention was prepared comprising 60 kg of palm kernel cake and 40 kg of water mixed with a combination of lactic acid bacteria (LAB) comprising Lactobacillus plantarum, Pediococcus acidilactici and Pediococcus pentosaceus.

[00173] Two other preparations (Sample 4 and Sample 5) are prepared according to the present invention, comprising:

[00174] - 60 kg palm kernel cake;

[00175] - 40 kg of water;

[00176] - a combination of lactic acid bacteria (LAB) Lactobacillus plantarum, Pediococcus acidilactici and Pediococcus pentosaceus; and Petition 870250084462, dated 09 / 19 / 2025, pp. 42 / 53 37 / 38

[00177] - the enzyme β-mannanase.

[00178] The ingredients of the various preparations (Samples 1-5) are well mixed, resulting in a solid-state fermentation of the palm kernel cake with a moisture content of about 40% and the temperature throughout the fermentation process is maintained at 40°C for 14 days / 336 hours (Samples 1-4) or 28 days / 672 hours (Sample 5). RESULTS:

[00179] The result of the fermentation is demonstrated by the solubility of the mannan-rich material, palm kernel cake, which is determined and quantified by high-performance liquid chromatography (HPLC), and the results show the following: TABLE 1 LAB included Enzymes included Fermentation time Solubility Sample 1 No No 0 hours (start) 14 days 15% 15% Sample 2 No Yes 0 hours (start) 14 hours 15% 22% Sample 3 Yes No 0 hours (start) 14 days 15% 31% Sample 4 Yes Yes 0 hours (start) 14 days 15% 35% Sample 5 Yes Yes 0 hours (start) 28 days 15% 48%

[00180] Thus, the results show a significant increase in the soluble mannan-oligosaccharide content of palm kernel cake when the palm kernel cake is subjected to Petition 870250084462, dated 09 / 19 / 2025, pages 43 / 53 38 / 38 fermentation. This increased solubility leads to greater digestibility of mannan-oligosaccharides and, consequently, of palm kernel cake when used, for example, in animal feed, such as feed for poultry or swine.

[00181] The results also show a significant increase in the soluble mannan-oligosaccharide content of palm kernel cake when the fermentation time is extended.

[00182] The initial test showed an insignificant increase in the solubility / digestibility of mannan-oligosaccharides and palm kernel cake after two days of fermentation, however after 3 days of fermentation a difference in solubility / digestibility was observed and it was significant after 14 days of fermentation and even more significant after 28 days of fermentation.

[00183] It is believed that the addition of additional enzymes such as glucanase, xylanase, cellulase, phosphatase or a combination thereof may further increase the solubility and digestibility of palm kernel cake. Petition 870250084462, dated 09 / 19 / 2025, pp. 44 / 53

Claims

1 / 3 CLAIMS 1. PROCESS FOR PROVIDING A PRODUCT WITH IMPROVED NUTRITIONAL VALUE FROM MANNAN-RICH MATERIAL, wherein the method is characterized by comprising the steps of: (i) providing the mannan-rich material; (ii) adding at least one microorganism to the mannan-rich material, providing an inoculated mannan-rich material; (iii) allowing the inoculated mannan-rich material to ferment for a period of at least 3 days, providing a fermented mannan-rich material with improved nutritional value.

2. PROCESS, according to claim 1, characterized in that at least one microorganism is selected from at least one lactic acid bacterial strain.

3. PROCESS, according to any one of claims 1 or 2, characterized in that at least one enzyme is added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii).

4. PROCESS, according to claim 3, characterized in that at least one enzyme added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii) is selected from the group consisting of glucanase (e.g., endo-1,3(4)-eglucanase), xylanase (e.g., endo-1,4-xylanase), cellulase, phosphatase (e.g., 6-phytase) and a combination thereof.

5. PROCESS, according to any one of claims 3 or 4, characterized in that at least one enzyme Petition 870250084462, dated 09 / 19 / 2025, page 45 / 53 2 / 3 added to the mannan-rich material in combination with at least one lactic acid bacterial strain in step (ii) being βmannanase (or β-mannosidase or β-D-mannoside mannohydrolase).

6. PROCESS, according to any of the preceding claims, characterized in that the mannan-rich material is selected from a palm cake material, a copra cake material, a guar germ cake material or a combination thereof, preferably the mannan-rich material is a palm cake material.

7. PROCESS, according to any of the preceding claims, characterized in that the fermentation process (i.e., step iii) does not include yeast fermentation and does not include fungal fermentation.

8. PROCESS, according to any of the preceding claims, characterized in that the moisture content of the inoculated mannan-rich material supplied in step (ii) and / or during the fermentation process in step (iii) can be in the range of 30-55% (w / w).

9. PROCESS, according to any of the preceding claims, characterized in that the inoculated mannan-rich material is fermented at a temperature in the range of 30-40 °C.

10. FERMENTED MANAM-RICH MATERIAL characterized by being obtained by the process as defined in any one of claims 1 to 9.

11. FERMENTED MANNAN-RICH MATERIAL characterized by comprising: - a crude protein content in the range of 5-30% (w / w); - a short-chain mannan oligomer; and - a lactic acid concentration of at least 200 mg / 100 g of fermented palm cake material.

12. FERMENTED MANNAN-RICH MATERIAL, according to any one of claims 10 or 11, characterized in that the short-chain mannan oligomer has a degree of polymerization (DP) in the range of 2-10, such as in the range of 2-8, for example, in the range of 2-6, such as in the range of 2-4.

13. MANNAN-RICH MATERIAL FERMENTED, according to any one of claims 10 to 12, characterized in that the solubility of the mannan and / or mannan-oligosaccharide-rich materials can be increased by at least 20%.

14. MANAN-RICH MATERIAL FERMENTED, according to any one of claims 10 to 13, characterized in that the digestibility of the mannan-rich material fermented is 50% or more, such as 55% or more, for example, 60% or more, such as 65% or more, for example, 70% or more, such as 75% or more, for example, 80% or more.

15. ANIMAL FEED PRODUCT characterized by comprising a material rich in fermented mannan, as defined in any one of claims 10 to 14. Petition 870250084462, dated 09 / 19 / 2025, pp. 47 / 53