A TWO-STAGE ENZYMATICALLY MODIFIED LEMON ALBEDO FLOUR PRODUCTION METHOD FOR GLUTEN-FREE BAKED GOODS.

Abstract

The invention relates to a two-stage enzymatically modified lemon albedo flour production method for gluten-free baked goods, developed to obtain a gluten-like elastic structure to solve problems such as hardness, loss of texture, low leavening, and brittleness encountered in gluten-free baked goods.

Description

A two-step enzymatic treatment for gluten-free baked goods. MODIFIED LEMON ALBEDO FLOUR PRODUCTION METHOD Technical Area The invention addresses the issues of hardness, loss of texture, and poor leavening encountered in gluten-free baked goods. Obtaining a gluten-like elastic structure to solve fragile structure problems Developed for this purpose and belonging to the field of food technology and functional nutrition, it is gluten-free. Double-stage enzymatically modified lemon albedo flour for baked goods It is related to the production method. State of the Art In gluten-free flour formulations with the known state of the art, elasticity and porosity are important. In order to provide structure, commonly: • Hydrocolloids: Hydroxypropyl methylcellulose (HPMC), xanthan gum, • Synthetic emulsifiers: Diacetyl tartaric acid ester of mono and diglycerides (Diacetyl tartaric acid ester of mono- and diglycerides, DATEM), • Single-step (or component-based) enzyme applications: Such as transglutaminase (TGase) These approaches are used. However, these approaches do not fully replicate the gluten-like elastic structure. is unable to provide this, requires high additive rates, and ensures uniformity in product texture. Losses are observed. Furthermore, it increases the need for unnatural additives and affects taste. It remains limited in terms of neutrality. On the other hand, some studies based on citrus albedo (e.g., US4526794A, US2023340164A1) enzymatic sequential even if enzymatic or drying processes have been applied Solutions including PME + TGas application and processing steps after flavonoid removal. This has not been explained. In other words, current techniques involve single-enzyme or just washing. includes drying processes. 1 Another known instance of the technique is called "Effect of Citrus Fiber on the Rheological “Properties of Dough and Quality of the Gluten-Free Bread (J. Korus et al., 2020)” The article discusses structural improvements in gluten-free bread using citrus fiber. This is a study aimed at providing this. This study examines the overall contribution of fibers, however specifically two-stage enzymatic modification of lemon albedo and pectin-protein It does not involve unique technical steps such as the creation of cross-links. Patent document number EP2548442, which falls under another known state of the art. This article discusses the use of transglutaminase (TGase) and its applications in gluten-free bread production. It is aimed at improving dough structure with TGas; that is, its invention is focused on this purpose. However This document describes the application of PME followed by TGas to lemon albedo, along with flavonoids. It does not involve a two-stage process based on pre-treatment of removal. The following studies examine different aspects of the topic (albedo / pectin content, citrus fiber effects, TGase-mediated protein cross-linking supports pectin-protein films. In their study, Korus et al. (2020) investigated the water retention, dough viscoelasticity, and properties of citrus fibers. This study analyzes the effects of citrus fiber on the final bread texture in gluten-free systems. It revealed that it partially improved the structure but did not create a gluten-like network. Bugarin et al. (2023) state that hydration in gluten-free breads with added citrus fiber is important. It shows improvement in fragility and tissue parameters. Additionally, citrus fiber has water-binding properties. It has been reported that lemon albedo affects shelf life with its capacity. Schröder et al. (2004) By demonstrating its high pectin content and gel-forming capacity, albedo's pectin Giosafatto et al. (2014) prove that pectin is a functional component rich in nutrients. and cross-linking of whey protein blends with TGase to form mechanically stable film structures. This shows that it can be formed. This mechanism involves pectin-protein interactions via TGase. It provides direct evidence that it can be strengthened. Moore et al. (2006) They studied the network-forming mechanism of transglutaminase in gluten-free bread. TGase forms cross-links between proteins in gluten-free systems, creating three gluten-like molecules. This is the fundamental study that explains its contribution to a multi-dimensional network structure. However, “lemon albedo (albedo flour) is first dimethylated with PME, then followed by TGaz By applying the mixture sequentially, pectin-protein covalent cross-links are formed, resulting in gluten-free products. There are no publications indicating that a gluten-like network is achieved in bread. 2 Therefore, with current techniques: • Two-step, sequential application of the enzyme combination, • Application of this enzymatic process after pre-treatment removal of bitter flavonoids. (PME + TGaz application), It is unknown and unpredictable by technical experts. Brief Description of the Invention The aim of the invention is to eliminate the hardness, loss of texture, and poor leavening issues encountered in gluten-free baked goods. and obtaining a gluten-like elastic structure to solve fragile structure problems Developed for this purpose and belonging to the field of food technology and functional nutrition, it is gluten-free. Double-stage enzymatically modified lemon albedo flour for baked goods The goal is to develop a production method. The invention involves modifying lemon albedo flour through special enzymatic and solvent processes. It is produced as an additive in gluten-free pastries (or baked goods) and A method of using gluten has been developed. Thanks to this, gluten-free bread, cakes and similar products can be made with gluten. Elasticity, volume, and porous structure are provided, improving the nutritional value and consumer acceptance of the product. is being increased. This invention involves the removal of flavonoids from lemon albedo flour first with an ethanol-water mixture (30–60%). this is done, followed by sequential administration of Pectin Methyl Esterase (PME) and Transglutaminase (TGase). It is based on the principle of processing with enzymes. - PME application creates free carboxyl groups in pectin chains. - Subsequently, covalent cross-links form between TGase and pectin-protein, forming gluten. A similar network structure is developed. The resulting modified albedo flour is added to gluten-free flour mixes at a rate of 5–15%. It is used. The type and sequence of enzymes used in the invention, and covalent cross-links between pectin and protein. Its purpose of forming and creating a gluten-like network structure is quite specific. 3 Advantages of the invention - Sequential enzymatic modification of lemon albedo with PME and TGase in two steps. As a result of processing, natural pectin-protein networks are formed in gluten-free dough. - Unlike known single enzyme applications, the enzyme sequence and processing conditions are different. This results in an unpredictable level of structural resilience. - Experimental results show a 25% increase in volume, a flexible, porous structure, and low brittleness. The results showed a significant improvement compared to the control group. - Natural, clean-label ingredients that produce gluten-like substances without the need for synthetic additives or emulsifiers. The structure is obtained. - Provides functional benefits. Lemon albedo is rich in fiber, pectin, and antioxidants. It increases the nutritional value of the product. - The products containing modified albedo flour obtained by the method described in the invention are taste-neutral and It is stable, and the flavor is preserved because bitter components are removed through flavonoid removal. It has no effect. - It does not contain synthetic emulsifiers. - The application is adaptable to industrial production. - The formulation in question does not contain synthetic emulsifiers. This is because it is stated in the description. such as, the invention is derived from naturally occurring sources (lemon albedo) and obtained through enzymatic modification. It uses an additive. Furthermore, one of the advantages of the invention is that it is synthetic. Gluten-like structure and elasticity without the need for emulsifiers or additives. This is ensured. In other words, gluten-free baked goods are made using modified lemon albedo flour. Elasticity and structural strength in products are obtained naturally and synthetically. The need for emulsifiers is eliminated. Detailed Description of the Invention The method described in the invention involves removing bitter flavonoids from lemon albedo. sequentially with pectin methylesterase (PME) and transglutaminase (TGase) enzymes It is based on processing. With PME application, free carboxyl groups are formed on pectin chains. groups are formed. Then, covalent bonds are formed between pectin and protein by applying TGas. Cross-links are formed, creating a natural network that mimics the elasticity of gluten. its structure is formed. 4 Hardness, loss of texture, poor leavening, and brittleness encountered in gluten-free baked goods. to obtain gluten-like elastic structures in order to solve the problems The improved invention concerns a method for obtaining modified lemon albedo-based flour as follows: The steps are as follows: - The first step is to separate the white part of the lemon peel (albedo), dry it, and turn it into flour. grinding to the right consistency, - In the second step, albedo flour is mixed with a predetermined ratio of ethanol-water. by washing to remove bitter flavonoids, - The third step is a primary enzymatic process, the Pectin Methyl Esterase (PME) process. Implementation: Within a predetermined temperature range and a predetermined... By applying pectin methylesterase for a period of time, the methyl ester groups of pectin are partially removed. hydrolysis and formation of free COO– groups - The fourth step is a second enzymatic process – the transglutaminase (Tgaz) process. Application: After the third step, transglutaminase is added to pectin and proteins. The process involves creating cross-links between the layers and obtaining modified albedo flour. The first step is to separate, dry, and grind the albedo, which is the white part of the lemon peel. Grinding processes to the desired consistency are the classic, generally known and widely used methods. It is based on methods. The parameters used in this stage are homogeneous in the product and This is critical for obtaining a stable albedo. In the albedo separation process, The lemon peel is completely separated from its outer yellow part (flavedo) and only The albedo portion is used. This distinction is important in terms of production efficiency and product quality. This is of great importance. During the drying stage, it is controlled at a temperature of 50–60 °C for 16–20 hours. A drying process is applied; this ensures that the moisture content of the albedo remains optimum. By reducing the level, the risk of microbial spoilage is decreased and enzymatic modification The appropriate physical structure is provided for these processes. The drying time and temperature are determined accordingly. Selecting the appropriate range is important for preserving the pectin structure and the quality of the final product. In the preferred application of the invention, the drying temperature is 55 °C and the drying time is 18 hours. It's been hours. The ethanol-water ratio mentioned in the second step is between 30% and 60%. Citrus peel and Removal of bitter flavonoids such as naringin and limonin in its albedo, in the literature It is usually made with ethanol-water mixtures ranging from 20–70%. The most commonly used range is 30– The answer is 60%. Because: flavonoid solubility decreases below 30%, and above 60% it increases. Pectin begins to dissolve and structural losses occur. Furthermore, in the second step... The 30–60% ethanol-water mixture ratios and washing frequency used for flavonoid removal are also... Effectively removing bitter-tasting components and leaving the product taste neutral. This has been determined experimentally. This pretreatment both increases the activity of the enzymes and This also prevents undesirable flavors in the final product. The flavonoid purification stage mentioned in the second step can be repeated 2-4 times. The Pectin Methyl Esterase (PME) treatment mentioned in the third step is performed at a pH range of 5.0–7.0. is being carried out. The treatment concentration of the PME enzyme mentioned in the third step is 10–40 U / g albedo. The dosage range has been selected. This dosage range ensures a controlled and effective enzymatic reaction. sufficient for this to occur, but possible side effects due to excessive enzyme use Reactions and cost increases have been avoided. In the third step, pectin methylesterase is involved in the formation of the free COO– groups mentioned. (PME) enzyme hydrolyzes the methyl ester groups in pectin, releasing free carboxylate (COO– This allows the release of COO– groups. These free COO– groups are then used in the next enzymatic step. It forms critical binding sites for cross-linking (e.g., with transglutaminase). In the third step, the PME enzyme mentioned is applied at a temperature of 35–45 °C for 60 minutes. PME application is generally performed at a temperature range of 35–45 °C and for a duration of 60 minutes. These conditions are applied and maximize enzyme activity while maintaining the pectin structure. It prevents unwanted degradation. The precise determination of these parameters ensures enzymatic processes. the success of the modification and the functional properties of the final product are critical to the invention. It is a technical detail that creates the step. The third step is added to albedo flour, which has been stripped of its flavonoids. In the preferred application of the invention, as the first enzymatic step in the first stage, at pH 6.0, 40 At °C temperature, pectin methyl esterase was administered at a dose of 20 U PME / g albedo flour for 60 minutes. (PME) has been applied. The processing concentration of the Transglutaminase (TGase) enzyme mentioned in the fourth step is 5– U / g albedo flour. In the preferred application of the invention, the second in the fourth stage. In the preferred application of the invention as the enzymatic step, at 37 °C for 45 minutes. Transglutaminase (TGaz) has been added to the albedo flour at a dose of U TGaz / g. The modified albedo flour obtained in the fourth step is pre-added to gluten-free flour mixes. It is added in a specified ratio. The modified albedo obtained in the fourth step. 6 Depending on the type of baking, 5–15% gluten is added to the gluten-free flour mix. The dough is then processed under standard fermentation and baking conditions. The modified albedo flour obtained using the method described in this invention can be used to make gluten-free bread, cakes, and pizza. The base can be used in the production of cookies or similar baked goods. Each parameter used in the method described in the invention relates to gluten in gluten-free baked goods. carefully selected and optimized to ensure optimal formation of a similar elastic structure. For example, in the third step, during the Pectin Methyl Esterase (PME) treatment, it is applied. Temperature, pH, and time values ​​affect the enzyme's demethylation activity on pectin. It is of critical importance because it has a direct impact. The PME enzyme is found in the 35–45 °C range and Application for approximately 60 minutes ensures sufficient free radicals in the pectin chains. This allows the carboxyl group to be exposed. These free COO– groups then follow, that is In the fourth step, during the Transglutaminase (TGase) process, strong bonds are formed between pectin and proteins. This is necessary for the formation of covalent cross-links. Temperature and time exceeding these limits... its release results either from insufficient enzyme activation or denaturation, thus reducing its effectiveness. It can lower the density of the gluten-like network, preventing its formation. Similarly, the temperature and enzyme dose selected for the TGas treatment also affect the gluten-free dough. It is optimized in terms of the effective establishment of cross-links. The preferred method of the invention. In the application, TGaz was administered at 37 °C for 45 minutes at a dose of 10 U / g albedo flour. It allows for the formation of ideal covalent bonds between protein and pectin. Higher High temperatures or excessive doses can impair the enzyme's activity or alter the dough's structure. It can have a negative impact. Problems that arise during the invention development process depending on parameter selection. It has been meticulously evaluated and resolved. For example, the PME application temperature is 50 °C. If it is removed, the enzyme's activity decreases, and consequently, pectin It was determined that insufficient carboxyl groups were forming in the chains. This problem is related to temperature and This was overcome by optimizing the pH range. Similarly, the TGaz dose being too high. If left untreated, undesirable hardening and structural deterioration of the dough have been observed. Therefore, the dosage range is limited. In conclusion, the method described in this invention involves the two-stage enzymatic extraction of lemon albedo. gluten-free breads are made using modified albedo flour produced through modification. Volume increase reached up to 25%, fragility decreased, and a porous structure was observed. 7 The product offered a texture comparable to its gluten-containing counterparts. It was obtained using the method described in the invention. The resulting final product can be marketed as a gluten-free food. This method involves enzymatic combinations and sequences unknown in current techniques. Because it is based on certain principles, it may not be readily predictable by a technical expert. Novel enzymatic sequencing-based TGase application following PME application. Sequential application in a way that creates a reaction mechanism, only the component not a combination, but a new one that alters the reaction dynamics and the microstructure of the product. a mechanism has been created. Thanks to this unique sequence of processes, gluten-free dough is made. Unpredictable levels of gluten-like network structure stability have been achieved. This effect The invention was not achieved through individual PME or TGaz applications in the literature. Therefore, the invention... It meets the invention step criterion; an innovative technique in gluten-free product technology. It offers progress. In conclusion, the methodological steps of the invention are as follows: - The order of enzyme application and processing conditions result in gluten-like, elastic results in gluten-free dough. and creates a robust network structure. - According to experimental results, the volume increase is up to 25%. - Fragility reduced by up to 30%, porous structure improved, textured gluten products. They have become close. - This effect is not achieved solely by using PME or TGaz alone. This is known in the literature. - The invention has developed a new reaction mechanism that is unpredictable for the expert. Example application: The subject of the invention is a sample application for the production of modified lemon albedo flour. The white part of the lemon peel (albedo) was separated, dried at 55 °C for 18 hours, and Albedo flour was obtained by grinding the material to pass through a 250 µm sieve. The resulting... albedo flour is mixed with a 40% ethanol–60% water mixture to remove bitter flavonoids. Mixed at a ratio of 1:10 (w / v), left to stand for 30 minutes, filtered, and this process repeated three times. This has been repeated. As a first enzymatic step, albedo flour, which has been purified of its flavonoids, was subjected to further processing. Pectin was infused at pH 6.0, 40 °C, at a dose of 20 U PME / g albedo flour, for 60 minutes. Methyl esterase (PME) was applied; immediately after this process, as a second enzymatic step... Transglutaminase (TGaz) at a dose of 10 U TGaz / g albedo flour for 45 minutes at 37 °C. added. After the process was completed, the product was dried at 50 °C, ground again, and 8 Modified lemon albedo flour is obtained by applying two-step enzymatic modification. It is a standard gluten-free product containing modified albedo flour, rice flour, and corn starch. The dough was prepared by adding 10% to the flour mixture, and the same formulation additive was used. It was created as a control sample without including the dough. The resulting doughs were kneaded using classic kneading techniques. It was processed under conditions of fermentation at 35 °C for 40 minutes and baking at 180 °C for 35 minutes. Industrial Applicability of the Invention The invention addresses the issues of hardness, loss of texture, and poor leavening encountered in gluten-free baked goods. Obtaining a gluten-like elastic structure to solve fragile structure problems Developed for this purpose and belonging to the field of food technology and functional nutrition, it is gluten-free. Double-stage enzymatically modified lemon albedo flour for baked goods This relates to the production method and includes natural, gluten-free bread, cake, croissant, and pizza bases. It can be used in products such as [mention specific products]. The invention is not limited to the above descriptions; a person skilled in the field can easily create the invention. It can present different applications. These are the claims of the invention and the protection sought. should be evaluated within this context. 9

Claims

1. Hardness, loss of texture, poor leavening, and brittleness encountered in gluten-free baked goods. Towards solving structural problems, aiming to obtain a gluten-like elastic structure. to be developed; - The first step is to separate the white part of the lemon peel (albedo), dry it, and turn it into flour. grinding to the right consistency, - In the second step, albedo flour is mixed with a predetermined ratio of ethanol-water. by washing to remove bitter flavonoids, - The third step is a primary enzymatic process, the Pectin Methyl Esterase (PME) process. Implementation: Within a predetermined temperature range and a predetermined... By applying pectin methylesterase for a period of time, the methyl ester groups of pectin are partially removed. hydrolysis and formation of free COO– groups - The fourth step is a second enzymatic process – the transglutaminase (Tgaz) process. Application: After the third step, transglutaminase is added to pectin and proteins. forming cross-links between them and obtaining modified albedo flour a modified lemon albedo-based flour production characterized by including the following steps method.

2. In the first step, the drying temperature mentioned is 50–60 °C and the drying time is 16–20 hours. A method like the one in claim 1, characterized by its nature.

3. The drying temperature mentioned in the first step is 55 °C and the drying time is 18 hours. A method like the one in claim 2, characterized by...

4. With the ethanol-water ratio mentioned in the second step being between 30% and 60%. A method like the one described in claim 3.

5. The flavonoid purification stage mentioned in the second step can be repeated 2-4 times. A method like the one in claim 4, characterized by...

6. The Pectin Methyl Esterase (PME) treatment mentioned in the third step is performed at a pH of 5.0–7.0 A method like the one in claim 5, characterized by being carried out within a specified range.

7. The treatment concentration of the PME enzyme mentioned in the third step is 10–40 U / g. A method like the one in claim 6, characterized by having albedo.

8. Application of the PME enzyme mentioned in the third step at a temperature of 35–45 °C for 60 minutes. A method like the one in claim 7, characterized by...

9. In the third step, as the first enzymatic step, 20 U is used at pH 6.0 and temperature of 40 °C. PPE / g albedo flour at a dose of pectin methyl esterase (PME) for 60 minutes. a method like the one in claim 8, characterized by its implementation.

10. The process of the Transglutaminase (TGase) enzyme mentioned in step four. claim 9, characterized by a concentration of 5–20 U / g albedo flour. a method like that.

11. In the fourth stage, as the second enzymatic step, 10 U was injected at 37 °C for 45 minutes. By adding transglutaminase (TGaz) enzyme to albedo flour at a dose of TGaz / g A method like the one described in claim 10. 11