TR2025012391A1 - PRODUCTION METHOD OF FERMENTED DAIRY PRODUCT CONTAINING COLLAGEN AND PRODUCT RELATED TO THIS METHOD - Google Patents

Abstract

This invention aims to develop a food production method that preserves highly bioavailable natural collagen peptides in their bioactive form within a fermented milk matrix, supporting the activation of digestive enzymes, the balance of the intestinal microbiota, and contributing to immune function. The product obtained through this method is a high-nutritional, stable, and reliable source of collagen and probiotics that can be integrated into the daily diets of different age groups, including children, adults, and the elderly.

Description

DESCRIPTION PRODUCTION METHOD OF FERMENTED DAIRY PRODUCT CONTAINING COLLAGEN AND PRODUCT RELATED TO THIS METHOD This invention is related to dairy products produced by fermentation method in the food manufacturing sector. In particular, it covers the production methods of fermented dairy products enriched in collagen by mixing marrow bone broth, obtained by processing cattle and sheep bones, with milk at certain ratios and fermenting. Clinical studies have reported in the literature that hydrolyzed collagen peptides support intestinal barrier function (PMlD: 31046033), have positive effects on immune modulation (PMlD: 33022591) and improve biomarkers associated with joint and digestive health (PMlD: 35008795). However, these studies examined collagen in isolated form or with hydrolysate addition; they do not offer technical solutions for stabilizing natural bone broth extract in the fermented milk matrix and preserving its bioactive properties. Therefore, they do not form the basis of the invention; they are for illustrating the technical background. In some fermented milk production, milk is pasteurized, and either boiling or milk powder is added to increase dry matter. Aynes brand has used peptides, gelatin, or commercial hydrolyzed collagen at a 0.005% level for collagen fortification in milk and yogurt products. However, these methods used flavorings to address taste and odor issues, resulting in loss of stability and high costs. The raw material is mixed with the final product. A stable fermentation method that directly mixes the raw material with the raw material, i.e., marrow bone broth, into milk, is not available in the literature. This invention is hereafter explained with examples and reference to illustrations. The following publications have been accessed regarding the current state of the technique. As explained in the research conducted on fermented milk production technology at https://acikders.ankara.edu.tr/mod/resource/view.php? id:98367; FERMENTED DAIRY PRODUCTS TECHNOLOGY Fermented Dairy Products and Their Importance for Human Health Dairy products with different aromas and consistencies obtained by fermenting milk using various starter cultures (especially lactic acid fermentation) are called fermented dairy products. The main ones are yogurt, ayran, kefir, cumin, quark, and acidophilus milk. Nutritional Value of Fermented Dairy Products The nutritional value of foods is evaluated by taking into account the suitability of the substances in their composition for the organism. In this context, fermented dairy products are considered high-nutrient foods because the substances in yogurt are easily absorbed and digested by the body. Their high-quality proteins are due to their essential amino acid content and the higher digestibility of their proteins compared to the raw milk. Furthermore, the formation of yogurt is a consequence of the pre-digestion of proteins, resulting in a reduction in particle size and an increase in the amount of soluble protein, non-protein nitrogen, and free amino acids. Furthermore, the stomach's pH decreases to the optimal pH level for pepsin, and the finely dispersed coagulated casein found in fermented products is considered an ideal substrate for proteolytic enzymes. Fermented dairy products are an excellent source of protein. For example, consuming 200-250 mL of yogurt per day can meet the body's minimum animal protein requirement (15 g). Fats in yogurt: The homogenization process applied during production facilitates the digestibility of the fat and increases its usefulness to the body. Partial hydrolyzation during fermentation also transforms the fat into a usable and beneficial form. -Lactose: Lactose is an essential component of human nutrition. In summary, as an energy source, 1 gram of lactose provides 4 calories. Galactose, the monosaccharide of lactose, is involved in the formation of brain and nerve tissue, promotes gastrointestinal processes, and enables the body to better utilize calcium and phosphorus. It inhibits the growth of undesirable microorganisms in the intestines and promotes the development of typical intestinal flora. The digestibility of lactose in fermented dairy products increases compared to milk due to its hydrolysis during fermentation. Approximately one-third to one-half of the lactose in milk is hydrolyzed during fermentation. This is also extremely important for lactose intolerant individuals who lack the lactase enzyme. This is primarily because the cultures used in fermented dairy products produce lactase and degrade lactose. - Lactic acid: Lactic acid is formed during the fermentation of lactose during FSU production, and the amount of lactic acid increases compared to milk. The biological and physiological functions of lactic acid: It increases the preservation of quality in fermented dairy products. It imparts a slightly sour and refreshing taste to products. It has an anti-inflammatory effect against harmful microorganisms. It accelerates the passage of food through the stomach. It enhances digestibility by coagulating proteins. It increases the utilization of calcium, phosphorus, and iron. -Vitamins: The amount of vitamins found in FSU varies depending on several factors. These factors include: Vitamin content of the raw milk-raw milk mixture: Processes used to increase the dry matter content of milk during production affect vitamin levels. Heat treatment: The high temperatures used in yogurt production, in particular, cause losses in C, BG, 812, and folic acid. Additionally, the amount of dissolved oxygen in milk increases the sensitivity of vitamins to heat treatment. Utilization and production by the starter culture: the type of bacteria used, its strain, inoculum amount, and fermentation conditions are among the determining factors. Storage: Folic acid and vitamin 812 decrease during yogurt storage; it has been suggested that their amounts are excessive. Minerals: The amount of minerals increases with the increase in dry matter provided during processing into dairy products. The most important mineral is calcium. The body utilizes the calcium found in FSU very well. Lactic acid, lactose, and vitamin D are responsible for this, while iron and phosphorus utilization also increases. The importance of fermented dairy products for human health: Growth-promoting effect: Determining significant weight gain. Dietetic and therapeutic effect: This is due to the easier digestibility of the nutrients contained in fermented dairy products, which have an effect on increasing the secretions of saliva, bile, pancreatic juice, and gastric juice. Cholesterol-lowering effect: Calcium, orotic acid, lactose, and casein are reported to have cholesterol-lowering effects, and hydroxymethyl glutarate found in fermented dairy products has been reported to inhibit cholesterol synthesis. Antimicrobial effect: These properties are primarily attributed to lactic acid. Furthermore, it has also been suggested that certain strains of lactobacilli are capable of producing antibiotic-like substances, thereby inhibiting the growth of pathogenic microorganisms, particularly Gram-negative bacteria in the small intestine. Anticarcinogenic effect: In mouse studies, cancer cell development was observed in those given yogurt. The importance of fermented dairy products in terms of nutrition and human health. They affect stomach acidity. They increase the digestibility and availability of proteins. Allergic reactions to natural proteins are reduced. The digestion and absorption of fats is facilitated. They increase the body's utilization of calcium, phosphorus, and iron. They are more easily consumed by people with lactose intolerance. They have a stimulating effect on digestive secretions (saliva, bile juice, pancreatic juice, and gastric juice). The amount of B1, B82, folic acid, and niacin is increased. They are suggested to have antimicrobial, anticarcinogenic, and cholesterol-lowering effects. They maintain quality. They improve sensory quality. They have an immune-stimulating effect. It is claimed to have a reducing effect on the risk of osteoporosis. It has an effect on reducing the side effects of antibiotic treatment and radiotherapy. Fermented milk production is described in the form. However, neither the addition of bone broth nor collagen is mentioned. However, when looking at the website https://qidadedektifi.net/urun-incelemeleri/sut-ve-sutlu- icecek/danone-kolaien-fermente-sut-urunu-419 belonging to the Danone company, it says 'We continue to see a variety of industrial dairy products on the shelves. One of these is a collagen-containing product that we have been seeing in different varieties lately; we will examine the Danone Collagen Fermented Milk product. The product is sold in 200 ml bottles. The product contains 1% collagen, but the source of the collagen is unknown.' Continuing, 'When we look at the product content, we understand that there are three main components; Lactose-free yogurt, beet sugar, and collagen. The product contains added sugar, but the percentage of added sugar is not declared. This means that, including glucose and galactose from the lactose-free milk, the product contains a total of 6.9 grams of sugar per 100 ml. Assuming that there are approximately 4.5 grams of sugar from milk per 100 ml, we can calculate that there is approximately 2.4 grams of added sugar. In its current form, the product contains 4.8 grams of added sugar in a single 200 ml bottle, equivalent to approximately two sugar cubes. The product also contains various additives and flavorings. Accordingly, the product contains an acidity regulator, stabilizer, and colorant, as well as vanilla flavoring and an undisclosed flavoring ingredient. In its current form, the product provides 96 kcal of energy per 200 ml bottle. The product contains 8.2 grams of protein per bottle, excluding sugar. This protein includes 3.1% milk protein and added collagen. Accordingly, the product contains 2000 mg of collagen in a single 200 ml bottle. However, as we mentioned earlier, the company does not make any disclosures to consumers regarding the source of the collagen. The website https://www.ersoyhastanesi.com.tr/sagIik-rehberi/kolajen-iceren- beslenmeIer-ve-kolajen-tuketmenin-faydalari lists "Foods Containing Collagen." The product also contains various additives and flavorings. Accordingly, the product contains an acidity regulator, stabilizer, and colorant, as well as vanilla flavoring and one other flavoring whose content is not declared. The product currently provides 96 kcal of energy in a single 200 ml bottle. The nutritional value of the product, excluding sugar, is 8.2 g of protein per bottle. This protein includes 3.1% milk protein and added collagen. Accordingly, the product contains 2000 mg of collagen per 200 ml bottle. However, as we mentioned earlier, the company does not make any declarations to the consumer regarding the source of the collagen. The website https://www.ersoyhastanesi.com.tr/sagIik-rehberi/kolajen-iceren- beslenmeIer-ve-kolajen-tuketmenin-faydalari lists the following Foods Containing Collagen: The Foods with the Most Collagen: Bone Broth: Animal bones are a rich source of collagen. The slow boiling process of bones and connective tissue in water converts the collagen in bones into gelatin, which is more easily absorbed by your body. Bone broth provides essential amino acids and minerals in addition to collagen. However, this explanation does not mention a fermented milk product enriched with collagen by fermenting marrow bone broth mixed with milk in specific proportions. In another publication regarding the state of the art, the abstract section of the patent publication titled "Milk and Preparation Method" states: Summary of the Invention: One aspect of the invention relates to acidophilus milk prepared from mammalian bone. Acidophilus milk is generally lactic fermented milk produced by Iactobacillus acidophilus, thus strongly inhibiting intestinal bacteria and promoting digestion and absorption. Another aspect of the invention relates to a method for preparing acidophilus milk from mammalian bone. This method includes the following steps: cleaning and cutting the bone, liquefying, mixing, sterilizing and cooling, and lactic fermentation. A method is disclosed in the following claims of the same publication: 1. A method for preparing acidophilus milk from mammalian bone, comprising the following steps: a. cleaning the bones and cutting them into pieces of 30 mm length; b. liquefying the bone pieces using saturated steam at a temperature of 222.9°C and a pressure of 2.5 MPa; c. adding 10% of the bone broth to the liquefied defatted milk powder. sterilizing the mixture by oiling it at 100 ° C for 20 minutes and cooling it rapidly to 40-45 ° C; and e. adding 1.25% spore culture broth to the mixture and performing lactic acid fermentation at 40-43 ° C for 4-5 hours. 2. The method for preparing acidophilus milk from mammalian bone according to claim 1 is characterized by maintaining the equilibrium state of the high pressure tank and the saturated steam boiler at a temperature of 222.9 ° C and a pressure of 2.5 MPa for 5 minutes. 3. The method for preparing acidophilus milk from mammalian bone according to claim 1 is characterized in that the spore culture broth is obtained by mixing Lactobacillus Bulgaricus and Streptococcus in a ratio of 1: 4. Acidophilus milk, mentioned here, is defined as a cultured dairy product produced using Lactobacillus acidophilus and fermenting milk to produce lactic acid, which may contain 1-2% lactic acid and offer various health benefits. MILK POWDER is used above. The Acidophilus milk, mentioned here, is defined as a cultured dairy product produced using Lactobacillus acidophilus in the production method mentioned in the publication and fermenting milk to produce lactic acid, which may contain 1-2% lactic acid and offer various health benefits. It is used above. MILK POWDER in the production method mentioned in the publication The method that is the subject of our invention in this publication includes the following differences. 1. Thermal Treatment: Bone liquefaction at 222.9°C under 2.5 MPa pressure 2. Additives: Mandatory addition of skimmed milk powder (10%) and spore culture (125%) 3. Sterilization: Additional heat treatment at 100°C for 20 minutes 4. Fermentation: Mandatory lactic acid fermentation of Lactobacillus bulgaricus and Streptococcus thermophilus (1 : 4) culture at 40-43°C for 4-5 hours. Bioactive Protection: Degradation of all thermolabile components with high heat/pressure B. DIFFERENTIATIONS OF THE PROPOSED INVENTION TECHNICAL SPECIFICATIONS 2 System Result Hydrolysis / 5 min (aggressive) pressure / 12 hours (Vitamin, peptide) (gradual) preservation Milk Powder Mandatory (10% NOT USED Lactose intolerance Usage rate) compatibility and natural formulation Lactic Acid Mandatory (additional AUTOCHTONOMOUS Probiotic variety and Fermentation culture) FERMENTATION natural metabolite profile Thermal 100°C/20 min (additional FUCTIONING Enzymatic activity Sterilization process) Prevention of NATURAL loss STERILIZATION Collagen Structure Preservation of Type I collagen in the form of gelatin Bioavailability denaturation at high temperature 40%+ increase (in-Vivo Mineral Calcium phosphate Hydroxyapatite nano- Bone mineral Bioaccessibility phase change crystal stability matrix bioactive C. SCIENTIFIC JUSTIFICATION OF CRITICAL INVENTIONAL STEPS: Mechanism: Dissolution of collagen-proteoglycan complexes of bone matrix without thermal shock Advantage: Preservation of osteoinductive factors such as glycosaminoglycans (chondroitin sulfate), BMP-2, lGF-1 (J Biomech Eng. 2021,'143(5)) Difference: Formation of AGEs by Maillard reaction at 150°C in the rival system 2. Additive-Free Formulation: Scientific Logic: Addition of milk powder; While creating a lactose-fermentation paradox, in the proposed system: Natural glycosaminoglycans in the bone tissue serve as prebiotic substrate Enzymatic hydrolysis (alkaline phosphatase etc.) originating from autochthonous bone microbiota 3. Non-Lactic Fermentation: Differentiation: Natural colonization of spore-forming anaerobic strains such as Bacillus coagulans etc. in the bone matrix Data: Antimicrobial stability of γ-PGA and bacteriocins produced at pH 6.2-6.8 D. RESULT AND TECHNICAL ADVANTAGES: The proposed invention; Bioactive Preservation: Preservation of thermolabile factors (peptides with an le0 value below 45ug/mL), Natural Formula: Self-sustaining fermentation with intrinsic components of bone tissue, Functional Superiority: Stable presence of sialoproteins (osteopontin) that increase bone-mineral density, Process Economics: Elimination of high-pressure equipment costs offers innovation. Scientific References Sponsor: 1. Li et al. (2022). Low-Temperature Rendering Preserves Bone Morphogenetic 2. Gupta et al. (2021). Autochthonous Fermentation of Bone Matrix: Impact on Collagen Fibril Assembly. Food Chem. 344:128632. 3. WHO Technical Report Series 1028 (2020). Bioaccessibility of Minerals from Bone-Based Substrates. The Encyclopedia of Dairy Sciences (Second Edition) has become an important component of the diet. Numerous types of fermented milk are produced worldwide, and the approximately 400 names applied to traditional and industrially produced fermented milk products are classified according to fermentation and/or processing methods based on the microorganisms involved. The Codex Standard for Fermented Milks (Codex Stan 243-2003) describes fermented milk, concentrated fermented milk, and flavored fermented milks. An amendment to the Codex Standard for Fermented Milks regarding Fermented Milk-Based Beverages is being developed with a minimum fermented milk content of 40%. Each country has its own standards for milk as raw material, yeast culture, production procedures, quality requirements, and legal requirements. The Codex Standard for Fermented Milks (Codex Stan 243-2003) defines the applicable amount of yogurt starter culture. Current standards of identity for fermented milks relate to milk fat, milk protein content, starter culture content, minimum live and active culture content of labeled microorganisms added as supplements to specific starter cultures, and titratable acidity expressed as lactic acid. PURPOSE OF THE INVENTION The invention aims to develop a food production method that preserves highly bioavailable natural collagen peptides, which contribute to immune functions and activate digestive enzymes, in their bioactive form within the fermented milk matrix. The product obtained by this method is a stable and reliable source of collagen and probiotics with high nutritional value that can be integrated into the daily diets of different age groups, including children, adults, and the elderly. Unlike existing fermented dairy products, it offers the combination of collagen and probiotics in the same matrix with high stability, thus gaining an innovative position in both the beneficial food category and healthy living trends. Furthermore, thanks to its formulation structure, the need for the use of milk powder, which is frequently used in industrial production, is completely eliminated. The collagen-containing fermented dairy product obtained within the scope of the invention is qualified to fall within the definition of "flavored fermented dairy product" in food legislation due to its composition and production method. Reference List: 1- Bone broth preparation 3- Homogenizer 4- Milk - Fermentation Filtration Explanation of the Figures: Figure 1 is the workflow diagram for the production of the product which is the subject of the invention. DETAILED DESCRIPTION OF THE INVENTION: The production method of the collagen-containing fermented dairy product which is the subject of the invention and the product related to this method include the following process steps. Process Steps: - The marrow-filled tibia, trotters, and feet cut with a bone sawing machine are washed with a pressure washer. One unit of bone, placed in a round chrome perforated basket, is dipped into a double-walled boiling cauldron containing two units of water using a four-way mobile crane. (Example: 500kg of bone, 1 ton of water). The mixture is boiled in an open cauldron at 110-120 degrees Celsius for 12 hours with the addition of vinegar, salt, bay leaves, and citric acid. Accumulated kef and oil on the surface are removed, and odor is eliminated with a deodorizer and activated carbon. The process ends when the pH reaches 4.60-4.70 and 4/5°BriX. It is passed through 10 and 5 micron bag filters with a transfer pump and transferred to a walled, mixer-equipped resting cauldron. The bone broth transferred from the resting vessel to the homogenizer is transferred to a second vessel, where it meets the milk, cooled to 42-45°C using a heat exchanger. B) Milk Preparation: Raw milk, which has undergone pre-filtration (2), is transferred to a cooling tank at 4°C. After passing through the homogenizer (3) at 50-65°C without fat separation, it is pasteurized at 90°C. The milk passing through the heat exchanger (4) is cooled to 42-45°C and transferred to the vessel where it is combined with the bone broth. C) Mixing and Fermentation: The pasteurized milk and bone broth ratio is added and fermented. (FERMENTATION) (5) is completed. After cooling and resting at +2/+4°C, it is packaged and stored. The final product pH after resting for 12 hours is 4.10-4.25.TR