Chitosan-based compositions with antimicrobial properties

Abstract

The present application is directed to antimicrobial compositions, comprising chitosan having an average molecular weight (Mw) of about 2 to about 100 kDa, an organic acid; and an additive selected from gum arabic, maltodextrin, gellan gum, guar gum, xanthan gum, psyllium, polyglutamate, carrageenan, pectin, alginate, hyaluronic acid, dextran, chondroitin, polyphosphate, and combinations thereof. The present application also includes a method of stabilizing and / or preserving a food or beverage product and a method of inhibiting growth of one or more microorganisms in the food or beverage product comprising adding an effective amount of one or more compositions of the application to the product.

Description

CHITOSAN-BASED COMPOSITIONS WITH ANTIMICROBIAL PROPERTIESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of priority from co-pending U.S. provisional patent application No. 63 / 689,915 filed on Septembers, 2024, the contents of which are incorporated by reference in their entirety.FIELD

[0002] The present invention relates to food or beverage antimicrobial compositions comprising chitosan and suitable additives which prevent or reduce the formation of insoluble chitosan aggregates when incorporated in the food or beverage products.BACKGROUND

[0003] Chitosan is a deacylated form of chitin, the second most abundant biopolymer on the planet after cellulose. Chitin is a long-chain polymer of N-acetylglucosamine, i.e., a structural analog of glucose in which the hydroxyl group on the C-2 site is replaced with the acetamido group (Fig. 1). Chitosan comes in a wide range of molecular masses (Mw). Most commercial chitosans have a Mw ranging from 50-2000 kDa, with an average degree of deacetylation (DDA) of 50-100%, commonly 80-90%, (Mourya et al. 2011 ). Due to the presence of primary amino groups, chitosan is soluble in dilute acids, such as acetic acid; its amino (ammonium) groups have a pKaof 6.5 (Mohammed et al, 2017). At acidic pH, these amines get protonated, become positively charged, and that causes chitosan to become a water-soluble cationic polyelectrolyte (Vida et al, 2015).

[0004] The source, method of extraction, and manufacturing processes have an impact on the properties of chitosan. The primary properties that determine the efficacy and scope of applications for chitosan include particle size, Mw, crystalline structure, %DDA, and surface area. Among these various properties, the Mw, polydispersity, crystallinity, and the pattern of distribution of N-acetylglucosamine units- 1 -10369140along the polymeric chain play roles in the physicochemical properties of chitosan (Kumirska, Weinhold, Thoming, & Stepnowski, 2011).

[0005] In addition, chitosan can be degraded by hydrogen peroxide treatment to obtain chitosan fractions of different Mw ranges such as, low, and medium Mw chitosan (Chang, Tai, & Cheng, 2001). Chitosan’s chain length and dispersion are thought to be one of the determinants of its biological activity. In contrast to the long chitosan strands, the low molecular mass chitosan (LMC) chains with a restricted distribution are easier to absorb on the surface of substrates. The LMC yields high solubility, aqueous solutions with little viscosity, and increased permeability, giving it a potential for use in the food, medical, and agricultural industries. Partial hydrolysis of chitosan shows that LMC has less crystalline structure than high molecular mass chitosan (HMC) (Chung, Yeh, & Tsai, 2011 ; Qin et al., 2002).

[0006] Chitosan exhibits antimicrobial activity and inhibits the growth of a wide variety of gram-positive and gram-negative bacteria, yeasts, and molds (Duan et al., 2019). The major mechanism of action of the antimicrobial activity of chitosan involves the interaction of chitosan in the cationic form with the anionic groups on the cell membrane of microorganisms, which may block the transport of essential substances to their interior or cause cellular rupture. Other hypothesized mechanisms of action involve the inhibition of RNA synthesis and disruption of protein synthesis in the bacterial cell membrane, causing disruptions that may lead to cell death; and chitosan forming complexation with metals, trace elements, and essential nutrients that are available for microorganisms growth, inhibiting the production of toxins (Muzzarelli et al., 2012; Zheng & Zhu, 2003). The antifungal activity of chitosan might be explained by the ability of chitosan to adsorb oxygen, since oxygen is essential for filamentous fungi and acetic acid bacteria (Rocha, Coimbra, & Nunes, 2017).

[0007] Chitosan has been applied as a preservative in various fruit juices, wine, and milk (Fernandes et al., 2008; Roller & Covill, 1999; Valera, Sainz, Mas, & Torija, 2017). However, the composition of the beverage may limit the antimicrobial activity of chitosan due to interaction and subsequent aggregation of chitosan with the beverage matrix.- 2 -10369140

[0008] Yoshida et al. (2020), described a tea beverage, which contains chitosan that does not lead to the formation of insoluble aggregates and the beverage does not get cloudy. However, according to this publication, formation of cloudiness can only be prevented when a non-fermented or semi-fermented tea beverage contains chitosan having a molecular weight of not more than 7 kDa at a concentration of not more than 200 ppm. In Yoshida, beverages containing chitosan having a molecular weight of 16 kDa or greater became cloudy. A bioactive complex formulation having compositions formed from natural ingredients, comprising chitosan, mineral salts, omega-3 rich oils, phospholipids, various casein derivatives, glycopeptides, phosphopeptides, etc. was described by Mora-Gutierrez et al. (2010).

[0009] A major challenge utilizing chitosan in various commercial applications, including food and beverages is the molecular interaction of chitosan with other ingredients / additives. The protonated amine binds to various anionic polysaccharides and forms electrostatic complexes that cause turbidity or aggregation in the food or beverage systems. The formation of such complexes affects the antimicrobial activity of chitosan, possibly by impairing its ability to interact with the cell membranes of the microorganisms.

[0010] Therefore, preventing or reducing chitosan aggregation in commercial applications such as food, beverages and the like is highly desired.SUMMARY

[0011] The present application discloses food and beverage antimicrobial compositions which include chitosan and a suitable additive. Upon incorporation of the composition in food and beverage products, interaction of the chitosan with other components of said food and beverage product is reduced, and thereby chitosan aggregation is reduced or prevented. As a result, the chitosan has higher antimicrobial activity in the product, resulting in enhanced stability of said product.

[0012] The present application relates to a food or beverage antimicrobial composition, comprising: chitosan having an average molecular weight (Mw) of about 2 to about 100 kDa; an organic acid; and- 3 -10369140an additive selected from gum arabic, maltodextrin, gellan gum, guar gum, xanthan gum, psyllium, polyglutamate, carrageenan, pectin, alginate, hyaluronic acid, dextran, chondroitin, polyphosphate, and combinations thereof.

[0013] In some embodiments, the additive is gum arabic.

[0014] In some embodiments, the average Mw of the chitosan is about 2 kDa to about 50 kDa. In some embodiments, the average Mw of the chitosan is about 50 kDa to about 100 kDa.

[0015] The present application also includes a food or beverage product comprising the composition of the application.

[0016] The present application also includes a method of stabilizing and / or preserving a food or beverage product comprising adding an effective amount of one or more compositions of the application to the product.

[0017] The present application also includes a method of inhibiting growth of one or more microorganisms in a food or beverage product comprising adding an effective amount of one or more compositions of the application to the product.

[0018] Other features and advantages of the present application will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating embodiments of the application, are given by way of illustration only and the scope of the claims should not be limited by these embodiments but should be given the broadest interpretation consistent with the description as a whole.DRAWINGS

[0019] The embodiments of the application will now be described in greater detail with reference to the attached drawings in which:

[0020] Figure 1 shows chemical structures of chitin (a), chitosan (b), and cellulose (c).

[0021] Figure 2 shows the core structure of polysaccharides of gum arabic (GA).- 4 -10369140

[0022] Figure 3 shows the optical density (OD) of the exemplary chitosan -based formulations in the cola-based beverages at 50 ppm dosage after 35 days of storage. Additive-1 is 1 % GA; additive-2 is 5% GA; additive 3 is 10% GA.

[0023] Figure 4 shows the effect of exemplary chitosan-based formulations on stability of cola-based beverages with A) control cola beverage (no chitosan or additives), B) medium molecular mass chitosan (MMC), and c) low molecular mass chitosan (LMC) at 50 ppm dosage, respectively, after 35 days of storage.

[0024] Figure 5 shows the OD of the exemplary chitosan-based formulations in orange juice-based beverages at 50 ppm dosage after 35 days of storage.

[0025] Figure 6 shows the effect of exemplary chitosan-based formulations on stability of orange juice-based beverages with A) control orange-juice beverage (no chitosan or additives), B) LMC, and C) LMC + Additive-2 at 50 ppm dosage, respectively, after 35 days of storage.

[0026] Figure 7 shows the OD of the exemplary chitosan-based formulations in the peach juice-based beverages at 100 ppm dosage after 35 days of storage.

[0027] Figure 8 shows the effect of exemplary chitosan-based formulations on stability of peach juice-based beverages with A) control peach juice beverage (no chitosan or additives), and B) MMC at 100 ppm dosage, respectively, after 35 days of storage.

[0028] Figure 9 shows the OD of exemplary chitosan-based formulations in the Ready to Drink (RTD) black tea-based beverages at 25 ppm dosage after 35 days of storage.

[0029] Figure 10 shows the effect of exemplary chitosan-based formulations on stability of RTD black tea-based beverages with A) control RTD black tea beverage (no chitosan or additives), B) MMC, C) LMC, and D) LMC + Additive-2 at 25 ppm dosage, respectively, after 35 days of storage.

[0030] Figure 11 shows the OD of the exemplary chitosan-based formulations in the cold coffee drink-based beverages at 50 ppm dosage after 35 days of storage.- 5 -10369140

[0031] Figure 12 shows the effect of the exemplary LMC chitosan-based formulations against A. niger in orange juice-based beverages at 50 ppm dosage.

[0032] Figure 13 shows the spoilage of the control (A) orange juice-based beverage and (B) mold inhibition activity of exemplary LMC chitosan-based formulation against A. niger in orange juice-based beverage at day 14 at 50 ppm dosage.

[0033] Figure 14 shows the effect of exemplary LMC chitosan-based formulations against A. niger in cola-based beverages at 50 ppm dosage.

[0034] Figure 15 shows the effect of exemplary LMC chitosan-based formulations against P. chrysogenum in cola-based beverages at 50 ppm dosage.

[0035] Figure 16 shows the SP values of the exemplary chitosan-based formulations in the orange juice-based beverages at 50 and 100 ppm dosage. Each bar for each dosage (50 ppm and 100 ppm) represents from left to right: control orange juice-based beverage, MMC, LMC, LMC+Additive 1 , LMC+Additive 2, and LMC+Additive 3.

[0036] Figure 17 shows the SP values of the exemplary chitosan-based formulations in the peach drink-based beverages at 100 and 200 ppm dosage. Each bar for each dosage (100 ppm and 200 ppm) represents from left to right: control peach drinkbased beverage, LMC, MMC, MMC+Additive 4 (1 % maltodextrin), MMC+Additive 5 (5% maltodextrin), and MMC+Additive 6 (10% maltodextrin).DESCRIPTION OF VARIOUS EMBODIMENTSI. Definitions

[0037] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present application herein described for which they are suitable as would be understood by a person skilled in the art.

[0038] The term “and / or” as used herein means that the listed items are present, or used, individually or in combination. In effect, this term means that “at least one of” or “one or more” of the listed items is used or present.- 6 -10369140

[0039] As used in the present application, the singular forms “a”, “an” and “the” include plural references unless the content clearly dictates otherwise. For example, an embodiment including “a microorganism” should be understood to present certain aspects with one microorganism species or two or more additional microorganisms.

[0040] In embodiments comprising an “additional” or “second” component, the second component as used herein is different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.

[0041] In understanding the scope of the present application, the term “comprising” and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, “including”, “having” and their derivatives.

[0042] The term “consisting” and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other unstated features, elements, components, groups, integers and / or steps.

[0043] The term “consisting essentially of”, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps.

[0044] The term "suitable" as used herein means that the selection of the particular compound or conditions would depend on the specific synthetic manipulation to be performed, and the identity of the molecule(s) to be transformed, but the selection would be well within the skill of a person trained in the art.

[0045] The terms "about", “substantially” and “approximately” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a - 7 -10369140deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies or unless the context suggests otherwise to a person skilled in the art.

[0046] The term “inhibit” or “reduce” and variations thereof as used herein means any detectable inhibition of a parameter in the presence of one or compositions of the application compared to otherwise the same conditions except in the absence of the one or more compositions of the application.

[0047] As used herein, the term “effective amount” means an amount of one or more compositions of the application that is effective to achieve the desired result. For example in the context of inhibiting microbial growth, an effective amount is an amount that, for example, increases said inhibition compared to the inhibition without administration of the one or more compositions.

[0048] The term “average molecular weight” or “Mw” as used herein refers to the weight average molecular weight as measured in kiloDaltons (kDa) using Size Exclusion Chromatography (SEC) or Gel Permeation Chromatography (GPC).

[0049] The term “composition of the application” or “composition of the present application” and the like as used herein refers to a composition comprising chitosan having an average molecular weight (Mw) of about 2 to about 50 kDa, an organic acid; and an additive as defined in the application.

[0050] When a range of values is presented in the present application, unless otherwise stated or understood, the range includes the end values (+ / - 5%) plus all values in 1 / 1 Oth increments therebetween.

[0051] The term “food or beverage product of the present application” or “food or beverage product of the application” as used herein refers to the food or beverage product comprising a composition of the application.II. Compositions of the Application- 8 -10369140

[0052] The present application includes a food or beverage antimicrobial composition, comprising: chitosan having an average molecular weight (Mw) of about 2 to about 100 kDa; an organic acid; and an additive selected from gum arabic, maltodextrin, gellan gum, guar gum, xanthan gum, psyllium, polyglutamate, carrageenan, pectin, alginate, hyaluronic acid, dextran, chondroitin, polyphosphate, and combinations thereof.

[0053] In some embodiments, the additive is gum arabic. In some embodiments, the additive is maltodextrin.

[0054] In some embodiments, the average Mw of the chitosan is about 2 kDa to about 80 kDa, about 20 kDa to about 60 kDa, about 30 kDa to about 50 kDa, or about 35 kDa to about 45 kDa. In some embodiments, the average Mw of the chitosan is about 2 kDa to about 50 kDa. In some embodiments, the average Mw of the chitosan is about2 kDa to about 20 kDa. In some embodiments, the average Mw of the chitosan is about 10 kDa to about 20 kDa. In some embodiments, the average Mw of the chitosan is about 50 kDa to about 100 kDa. In some embodiments, the average Mw of the chitosan is about 60 kDa to about 90 kDa. In some embodiments, the average Mw of the chitosan is about 70 kDa to about 80 kDa.

[0055] In some embodiments, the composition further comprises water and is a solution.

[0056] In some embodiments, the chitosan is present in the composition in an amount of about 2 % w / v to about 10% w / v based on the total volume of the composition. In some embodiments, the chitosan is present in the composition in an amount of about3 % w / v to about 8 % w / v based on the total volume of the composition. In some embodiments, the chitosan is present in the composition in an amount of about 5% w / v based on the total volume of the composition.

[0057] In some embodiments, the chitosan has a degree of deacetylation that is about 50% or greater than 50%. In some embodiments, the degree of deacetylation of the chitosan is about 80% or greater than 80%.- 9 -10369140

[0058] In some embodiments, the additive is present in the composition in an amount of about 0.5 % w / v to about 20 % w / v based on the total volume of the composition. In some embodiments, the additive is present in the composition in an amount of about 0.8 % w / v to about 12 % w / v based on the total volume of the composition. In some embodiments, the additive is present in the composition in an amount of about 1 % w / v, about 5% w / v, or about 10 % w / v based on the total volume of the composition. In some embodiments, the additive is present in the composition in an amount of about 5 % w / v to about 10 % w / v based on the total volume of the composition.

[0059] In some embodiments, the organic acid is selected from citric, acetic, malic, formic, propionic and tartaric acids. In some embodiments, the organic acid is citric acid. In some embodiments, the organic acid is acetic acid.

[0060] In some embodiments, the organic acid is present in the composition in an amount of about 1 % w / v to about 15 % w / v based on the total volume of the composition. In some embodiments, the organic acid is present in the composition in an amount of about 1 % w / v, about 5 % w / v, about 10 % w / v or about 15 % w / v based on the total volume of the composition.

[0061] In some embodiments, the composition of the application is in the form of a liquid solution or suspension. In some embodiments, the composition of the application is in the form of an aqueous solution. In some embodiments, the composition of the application is in the form of a powder, obtained by drying a composition of the application to obtain the powder.

[0062] In some embodiments, when the composition of the application is in a dry form, the chitosan is present in the composition in an amount of about 10 % w / w to about 15% w / w, the organic acid is present in the composition in an amount of about 15 % w / w to about 20 % w / w, and the additive is present in the composition in an amount of about 25 % w / w to about 35 % w / w, based on the total weight of the dry composition.

[0063] In some embodiments, the composition of the application comprises further ingredients and excipients known in the art. Exemplary ingredients / excipients include but are not limited to pigments, colorants, phenolic acids, inorganic anions, emulsion- 10 -10369140stabilizers, and carbohydrate additives (alginate, xanthan gum, carrageenan, starch, pectin, etc.). In some embodiments, the composition is formulated for inclusion in a specific product and ingredients are added to facilitate this inclusion as would be known to a person skilled in the art. In some embodiments, when the composition of the application is in the form of a liquid solution or suspension, the balance of the composition is water.

[0064] The present application also includes a food or beverage product which comprises the composition of the present application. The product may be in any form, including liquid, semi-solid and solid and combinations thereof.

[0065] Accordingly, the present application further includes a food or beverage product comprising one or more compositions of the application.

[0066] In some embodiments, the composition is present in the food or beverage product in an amount of about 10 ppm to about 400 ppm. In some embodiments, the composition is present in the food or beverage product in an amount of about 25 ppm to about 200 ppm. In some embodiments, the composition is present in the food or beverage product in an amount of about 25 ppm, about 50 ppm, about 75 ppm, about 100 ppm, about 150 ppm, or about 200 ppm.

[0067] In some embodiments, the composition of the present application substantially reduces or prevents formation of insoluble chitosan aggregates in the food or beverage product of the present application compared to formation of insoluble chitosan aggregates in the food or beverage product without the composition of the present application. Thus, in some embodiments, the food or beverage product of the present application has little or no sedimentation or aggregation.

[0068] In some embodiments, the chitosan and the additive of the application form molecular assemblies, such as, but not limited to a complex coacervate system of chitosan and the additive.

[0069] In some embodiments, the composition of the application present in the food or beverage increases the preservation time of said food or beverage product. Thus, in some embodiments, the food or beverage product of the present application has a - 11 -10369140preservation time at room temperature that is increased by at least 10%, by at least 30% or by at least 50%, compared to a preservation time of the food or beverage product without the composition of the present application. As such, the composition of the application present in the food or beverage increases the stability of the product of the present application.

[0070] In some embodiments, the composition of the application present in the food or beverage inhibits the growth of microorganisms in said food or beverage product. In some embodiments, the composition of the present application inhibits the growth of one or more microorganisms selected from bacteria, yeast and mold.

[0071] In some embodiments, the growth of one or more microorganisms in the food or beverage product is inhibited by at least 20% compared to the growth of the one or more microorganisms in the food or beverage product without the composition of the present application. In some embodiments, the growth of the species is inhibited by at least 30%, by at least 40%, or by at least 50%.

[0072] In some embodiments, the microorganism is a gram-negative bacteria. In some embodiments, the gram-negative bacteria is Escherichia coli (E. coli).

[0073] In some embodiments, the microorganism is a yeast. In some embodiments, the yeast is Zygosaccharomyces baili (Z. baili).

[0074] In some embodiments, the microorganism is a mold. In some embodiments, the mold is Aspergillus niger (A. niger). In some embodiments, the mold is Penicillium chrysogenum (P. chrysogenum).

[0075] In some embodiments, the food or beverage product of the present application is a beverage product. In some embodiments, the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juicecontaining beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks. In some embodiments, the beverage is orange juice, orange-juice containing beverage, peach juice, peach juicecontaining beverage, tea, tea- containing beverages such as ready to drink black tea-- 12 -10369140containing beverages and the like, or coffee-containing beverages such as cold coffee drink- containing beverages and the like.

[0076] In some embodiments, the beverage product further comprises one or more of glycerol, propylene glycol, and ethylenediaminetetraacetic acid.

[0077] In some embodiments, the composition of the present application reduces or prevents the turbidity of the beverage product compared to the beverage product without the composition of the present application This effect is achieved due to the reduction or prevention of the formation of insoluble chitosan aggregates in the food or beverage product.III. Methods of Using the Compositions of the Application

[0078] The present application includes a method of stabilizing and / or preserving a food or beverage product comprising adding an effective amount of one or more compositions of the present application to the product.

[0079] In some embodiments, the food or beverage product is stabilized and / or preserved at room temperature by at least 10%, by at least 30% or by at least 50%, longer than a food or beverage product without the one or more compositions of the application.

[0080] In some embodiments, the food or beverage product is a beverage product. In some embodiments, the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juice-containing beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks. In some embodiments, the beverage is orange juice, orange-juice containing beverage, peach juice, peach juice-containing beverage, tea, tea-containing beverages such as ready to drink black tea-containing beverages and the like, or coffee- containing beverages such as cold coffee drink-containing beverages and the like.

[0081] In some embodiments, the effective amount of the one or more compositions of the present application for stabilizing and / or preserving the food or beverage - 13 -10369140product is about 10 ppm to about 400 ppm. In some embodiments, the effective amount of the one or more compositions is about 25 ppm to about 200 ppm. In some embodiments, the effective amount of the one or more compositions is about 25 ppm, about 50 ppm, about 75 ppm, about 100 ppm, about 150 ppm, or about 200 ppm.

[0082] The present application also includes a method of inhibiting growth of one or more microorganisms in a food or beverage product comprising an effective amount of one or more compositions of the present application to the product.

[0083] In some embodiments, the growth of the one or more microorganisms is inhibited in the food or beverage product by at least 20% compared to the growth of the one or more microorganisms in the food or beverage product without the composition of the present application. In some embodiments, the growth of the species is inhibited by at least 30%, by at least 40%, or by at least 50%.

[0084] In some embodiments, one or more microorganisms are selected from bacteria, yeast and mold.

[0085] In some embodiments, the microorganism is a gram-negative bacteria.

[0086] In some embodiments, the gram-negative bacteria is Escherichia coli (E. coi).

[0087] In some embodiments, the microorganism is a yeast.

[0088] In some embodiments, the yeast is Zygosaccharomyces baili (Z. baili).

[0089] In some embodiments, the microorganism is a mold.

[0090] In some embodiments, the mold is Aspergillus niger (A. niger). In some embodiments, the mold is Penicillium chrysogenum (P. chrysogenum).

[0091] In some embodiments, the food or beverage product is a beverage product. In some embodiments, the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juice-containing beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks. In some embodiments, the beverage is orange juice, orange-juice containing beverage, peach juice, peach juice-containing beverage, tea, - 14 -10369140tea-containing beverages such as ready to drink black tea-containing beverages and the like, or coffee- containing beverages such as cold coffee drink-containing beverages and the like.

[0092] In some embodiments, the effective amount of the one or more compositions of the present application for inhibiting the growth of the one or more microorganisms is about 10 ppm to about 400 ppm. In some embodiments, the effective amount of the one or more compositions is about 25 ppm to about 200 ppm. In some embodiments, the effective amount of the one or more compositions is about 25 ppm, about 50 ppm, about 75 ppm, about 100 ppm, about 150 ppm, or about 200 ppm.

[0093] The present application also includes a method of substantially reducing or preventing the formation of insoluble chitosan aggregates in the food or beverage product comprising adding an effective amount of one or more compositions of the present application to the product. Thus, in some embodiments, the food or beverage product of the present application has little or no sedimentation or aggregation. Also, in some embodiments, due to the reduction in formation of insoluble chitosan aggregates, turbidity of the product is reduced or prevented.

[0094] In some embodiments, the food or beverage product is a beverage product. In some embodiments, the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juice-containing beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks. In some embodiments, the beverage is orange juice, orange-juice containing beverage, peach juice, peach juice-containing beverage, tea, tea-containing beverages such as ready to drink black tea-containing beverages and the like, or coffee- containing beverages such as cold coffee drink-containing beverages and the like.

[0095] In some embodiments, the effective amount of the one or more compositions of the present application to substantially reduce or prevent the formation of insoluble chitosan aggregates in the food or beverage product is about 10 ppm to about 400 ppm. In some embodiments, the effective amount of the one or more compositions is about 25 ppm to about 200 ppm. In some embodiments, the effective amount of the- 15 -10369140one or more compositions is about 25 ppm, about 50 ppm, about 75 ppm, about 100 ppm, about 150 ppm, or about 200 ppm.IV. Methods of Preparing the Compositions of the Application

[0096] The application further includes a method of making the compositions of the application. In some embodiments, the method comprises: combining chitosan having an average molecular weight (Mw) of about 2 to about 100 kDa, an organic acid; and an additive selected from gum arabic, maltodextrin, gellan gum, guar gum, xanthan gum, psyllium, polyglutamate, carrageenan, pectin, alginate, hyaluronic acid, dextran, chondroitin, polyphosphate, and combinations thereof. In some embodiments, the composition of the application is in the form of an aqueous solution.

[0097] The food or beverage product containing the composition of the application may be obtained by combining one or more compositions of the application with the final product. In some embodiments, the compositions of the application are incorporated as an ingredient or a component during the preparation of the product.

[0098] In some embodiments, the composition of the application is added to the food or beverage product in the form of a liquid solution or suspension. In some embodiments, the composition of the application is added to the food or beverage product in the form of a powder, obtained by drying the composition of the application to obtain the powder.

[0099] It is appreciated that the food or beverage product may include further ingredients and excipients known in the art. Exemplary ingredients / excipients include but are not limited to pigments, colorants, phenolic acids, inorganic anions, emulsion stabilizers, and carbohydrate additives (alginate, xanthan gum, carrageenan, starch, pectin, etc.).

[0100] The chitosan having an average molecular weight (Mw) of about 2 to about 100 kDa can be obtained by any method known in the art from a fungus, crustacean or an insect.- 16 -10369140EXAMPLES

[0101] Amongst various Food Grade (FG) additive materials, gum arabic (GA) (shown on Fig. 2) is widely used in different food applications. It has high water solubility, and its low viscosity can create a protective film around core materials, and act like an emulsifier to prevent aggregation by forming a thick layer. Similarly, maltodextrin has low viscosity, good solubility even at high concentrations, and is commonly used for the encapsulation of natural extracts / flavors, and as a carrier material for spray drying.

[0102] Gum arabic (gum acacia), such as for example CAS Registry Number: 9000-01-05, has a Mw of about 250 kDa, and its solutions are slightly acidic, around pH 4.5 - 5.5. According to the definition of the Joint Expert Committee for Food Additives (JECFA), GA is a dried exudate obtained from the stem and branches of acacia trees. GA is a natural branched-chain multifunctional hydrocolloid with a highly neutral or slightly acidic, high molecular weight arabino-galactan-protein complex containing calcium, magnesium, and potassium salts, which on hydrolysis yields three main fractions of polysaccharides and proteins, including arabinogalactan peptide, arabinogalactan protein, and glycoprotein. These fractions contribute to about 85-90, 10, and 2% of total mass and contain approximately <1 %, -10%, and 25-50% proteinaceous materials, respectively (Musa, Ahmed, & Musa, 2019). The arabinogalactan peptide and arabinogalactan protein have a thin oblate ellipsoid or disk-like highly branched carbohydrate blocks attached to a polypeptide chain and the glycoprotein contains a mixture of spheroidal ring-like structures. The hydrophobic protein component functions as an emulsifier which adsorbs onto surface of oil droplets, while hydrophilic carbohydrate component inhibits flocculation and coalescence of molecules through electrostatic and steric repulsions in food additives (Sabet et al., 2021 ).

[0103] Maltodextrins (MD), such as for example, CAS Registry Number: 9050- 36-06, Mw of about 1-2 kDa, are hydrolysis products of starch. Maltodextrin is obtained by acid and / or controlled enzymatic hydrolysis of starch and consists of a-(1 ,4) linked D-glucose units. Maltodextrin contains 2-3 % glucose and 5-7 % maltose and can be made into a white hygroscopic spray-dry powder (Kibici & Kahveci, 2019). The extent - 17 -10369140of starch degradation in MD is indicated by the dextrose equivalent value (DE) that evaluates the content of reducing-end groups, which is the inverse value of the average degree of polymerization (DP) of dehydrated glucose units. The DE value of MD is in the range of 3-20, indicating that its carbohydrate chain is long, and it is a complex mixture of high and low molecular substances (Garnero, Aloisio, & Longhi, 2013).

[0104] Maltodextrin is widely used in the food industry as a coating, carrier, and as stabilizer material due to its high solubility, low viscosity at high solids concentration, low relative cost, neutral taste, and high thermal and freeze stability. Maltodextrin can have host-guest interaction with another polysaccharide like chitosan. According to the molecular size of the core material, the conformation of MD changes from a flexible coil to a helix shape. The C1 and C4 carbons are more sensitive to conformational changes in the presence of a core polysaccharide (Garnero et al., 2013). FT-IR spectroscopy and1H-NMR nuclear magnetic resonancebased studies have demonstrated that amino and carbonyl groups in a polysaccharide can be complexed with MD through dispersion force, and may form hydrogen bonds with the hydroxyl group of pyranose sugar of MD (Kalra, Bhat, & Kaur, 2021 ; Xiao, Xia, Zhao, Niu, & Zhao, 2022).

[0105] The following non-limiting examples are illustrative of the present application:Example 1 - Preparation of the Base Chitosan-based Preservative Formulations

[0106] Two grams of chitosan fractions (low or medium Mw) were taken in a 50 mL screw cap conical tubes (Sarstedt Canada, Montreal, QC) and dissolved in a 40 mL aqueous solution of organic acids such as, citric, acetic or malic acids (2.8 g of organic acid in water). The formulation was then mixed in an Elmi Sky Line Analog Orbital Shaker (Cole-Parmer, Quebec city, QC) for 2-3 hours at room temperature to allow complete mixing of the chitosan.Example 2 - Preparation of Chitosan-based Additive Formulations- 18 -10369140

[0107] One to ten percent w / v of different additives such as, gum arabic (GA), maltodextrin (MD), alginate or others were directly incorporated in a 50 mL screw cap conical tubes (Sarstedt Canada, Montreal, QC) containing the base liquid chitosan based preservative formulations (low or medium Mw). The formulations were then mixed in an Elmi Sky Line Analog Orbital Shaker (Cole-Parmer, Quebec City, QC) for 2-3 hours and were monitored to obtain a completely dissolved formulation. Exemplary Additives used in the present application are as follows: Additive 1 - 1 % GA; Additive 2 -5% GA; Additive 3 -10% GA; Additive 4: 1 % Maltodextrin; Additive 5: 5% Maltodextrin; Additive 6: 10% Maltodextrin; GA used in the examples was Gum arabic (gum acacia), CAS Registry Number: 9000-01-05 with a Mw of about 250 kDa. MD used in the examples was CAS Registry Number: 9050-36-06 with a Mw of about 1-2 kDa.Example 3 - Testing of Preservative Formulations in Beverages

[0108] Both, the chitosan-based preservative formulations without additives and the chitosan-based formulations with additives were tested in various beverages. For example, 25 to 400 ppm of formulations were drawn from the conical tubes with a pipette (Thermo Fisher Scientific Canada) and incorporated into a conical tube containing 25 mL of the beverage being tested, followed by manual shaking for 30 to 60s at room temperature. The beverages were kept at room temperature and checked for level of aggregation / sedimentation (visual appearance, optical density, streaming potential, etc.) over a span of 1 to 35 days.Example 4 - Measurement of Optical Density

[0109] One milliliter of beverage sample was taken with a pipette (Thermo Fisher Scientific Canada) and placed into a 4.2 mL polystyrene cuvette (Sarstedt Canada, Montreal, QC). The optical density of the beverages was tested on an UV- visible spectrometer (VWR UV-3100PC) equipped with the UV Analyst software using a quartz flow cell with 1 mm of optical path. The spectra were measured using the transmittance technique at 595 nm.- 19 -10369140Example 5 - Measurement of Streaming Potential

[0110] The streaming potential as a result of streaming current is one of the characteristics of electrolyte-containing liquids (Q. Chang, 2016). A streaming current is an electric current which originates when the electrolyte solution is flowing in a chamber under a pressure gradient. If the surface of the chamber is charged, an electrical double layer develops a local increase in the counterion concentration and a local decrease in the concentration of co-ions (ions of the same sign as the surface charge). Upon relative movement of the liquid with respect to the solid sample, the ions of the electrochemical double-layer are sheared off their equilibrium position and shifted along the solid surface. The double layer develops over the order of the Debye screening length, which is ionic strength (salt concentration) dependent. The transport of these charges in the double layer generates a current and a concomitant potential difference between the entrance and the exit of the chamber. If the measured potential is unequal to zero, the sign of the measured value merely indicates whether the charge is positive or negative (Olthuis, Schippers, Eijkel, & van den Berg, 2005). Streaming potential depends on the constants that characterize the macroscopic behavior of the solution and the surface-solution microscopic interactions. Therefore, the streaming potential values of a beverage can be affected and vary by the viscosity, the ionic diffusivities, the dielectric constant, the surface charge density (or the zeta potential), and the carbonation level (Oatley-Radcliffe, Aljohani, Williams, & Hilal, 2017).

[0111] Streaming potential was found to be very useful during the studies shown in the present application. In addition, it could be used in the future to calculate the zeta potential and these measurements could become a characterization method for surface functionality or stability of dispersed particles in the systems described in this application.

[0112] The streaming potential (SP) of the beverages was measured after 1 day using a particle charge detector (PCD-03, BTG Mutek GmbH). The central element was a plastic measuring cell with a fitted displacement piston. Approximately 10-15 mL of the beverage sample was filled into a polytetrafluoroethylene measuring cell at room temperature where colloidally dispersed molecules adsorbed on the plastic surface of the piston and on the cell walls via van der Waal forces. The counter-ions - 20 -10369140remained comparatively free. A defined narrow gap was allowed between cell wall and piston. Driven by a motor, the piston oscillates in the measuring cell and creates an intensive liquid flow which entrains the free counter-ions, thus separating them from the adsorbed sample material. At the built-in electrodes, the counter-ions induce a current which is rectified and amplified electronically. The streaming potential was measured in mV shown on the display with the appropriate sign.Example 6 - Inoculum Preparation

[0113] The Z. baili and E. coli were cultivated in potato dextrose and tryptic soy broth (TSB), respectively, at 25-30°C for 2 days. The cell density of the cultures were estimated by measuring the optical density of the cultures. The cultures were diluted in buffered peptone water pH 7.4 (BPW) to produce a suspension containing approximately 5 x 104cells per mL. These suspensions were used as inoculum (1 % v / v) in the challenge tests. For the mold challenge test, Aspergillus niger (A. niger) and Penicillium chrysogenum (P. chrysogenum) were grown on potato dextrose broth or agar plate at 25-30°C for 5 days. Spores were collected from the tubes or plates in a suspension by adding 9 mL of BPW containing 10 pl of Tween-20 to the tube and subsequently vortexing vigorously. The density of the spore suspension was estimated based on microscopic observation. The spore suspension was diluted in BPW to produce a suspension containing approximately 5 x 104spores per ml. These spore suspensions were used as inoculum in the challenge tests.Example 7 - Challenge Test

[0114] Beverages were supplemented with the appropriate dose of chitosan- based formulations and were mixed well. A 10 to 100 mL of the beverage sample was transferred to tubes or jars, inoculated with the target microorganisms (final density of 103cells per ml), mixed well, and stored at 25-30°C. During the incubation, samples were taken to determine the density of viable cells by plate counting and the results were expressed as Log CFU / mL.- 21 -10369140Example 8 - Visual Mold Challenge Test

[0115] A 25 mL of the beverage sample was transferred to tubes and was supplemented with the appropriate dose of chitosan-based formulations and mixed well. The beverage samples were then inoculated with a 25-30 pl of the target microorganism (A. niger and P. chrysogenurrf) to obtain approximately 100 spores per ml of inoculation level, mixed well, and stored at ambient temperature (20-25°C) for visual observation of mold growth. The beverages were observed every day and were considered spoiled if any visible signs of mold growth were detected. The spoilage day is presented as the difference between initial inoculation and number of days to spoilage. The test was carried out for 60 days.RESULTSEffect of Chitosan fractions on stability of Cola-based Beverages

[0116] Chitosan fractions-based formulations demonstrated a dose and Mw- dependent stability in the beverages tested. In a low molecular mass chitosan (LMC) formulation for beverage treatment, characterized by a Mw ranging from 2 to 50 kDa and more suitably 2 to 20 kDa, and a degree of deacetylation (DDA) greater than 50, suitably greater than 80, demonstrated high stability in cola-based beverages (Figure 3) at a dosage of 50 ppm . After 35 days of storage the OD of the control cola beverage tested was 0.4 to 0.45. At 50 ppm dosage, the OD of cola beverage with a medium molecular mass chitosan (MMC) formulation for beverage treatment, characterized by a Mw ranging from 50 to 100 kDa, and DDA greater than 50, suitably greater than 80, was found to be 0.05 to 0.11 , and there was visible sedimentation probably due to aggregation / interaction, observed in the beverage (Figure 4B). Whereas the cola beverage containing 50 ppm of LMC was found to be stable with little or no sedimentation (Figure 4C) and the OD was found to be 0.4 to 0.45 (Figure 3), which was much higher than the OD of the MMC and similar to that of the control.Effect of Chitosan Fractions and Gum Arabic (GA) Additive on Stability of Orange Juice-based Beverages- 22 -10369140

[0117] The Mw of chitosan and also incorporation of GA as an additive significantly influenced the stability of the orange juice-based beverages. After 35 days of storage the OD of the control orange juice-based beverage was tested to be 0.35 to 0.38. At 50 ppm dosage, the OD of the beverage with MMC was 0.06 to 0.10 and the OD of the LMC was 0.2 to 0.25, which was significantly higher than the MMC (Figure 5). Incorporation of GA had positive effects on the stability of the orange juicebased beverages. At 50 ppm dosage and after 35 days, the OD of the LMC+Additive- 2 (5% GA) was found to be 0.45 to 0.5, which is almost 2 times higher than the OD of the LMC without GA (at the same dosage) and also higher than OD of the control after 35 days (Figure 5). Visible sedimentation probably due to aggregation / interaction and phase separation was observed in the beverage containing only LMC (Figure 6B). The beverage containing 50 ppm of LMC+Additive-2 (Figure 6C) was found to be stable and similar to that of the control orange juice (Figure 6A).Effect of Chitosan Fractions on Stability of Peach Juice-based Beverages

[0118] The MW of chitosan significantly influenced the stability of the peach juice based beverages. The MMC formulation demonstrated better stability in peach drink than that of the LMC formulation. For example, after 35 days of storage the OD of the control peach juice beverage tested was 0.18 to 0.22 (Figure 7). The OD of the peach juice beverage with LMC was found to be 0.05 to 0.1 , whereas the OD of the beverages with MMC was 0.23 to 0.28, which was higher than the control. Visually the peach drink with 100 ppm of MMC looked very similar to that of the control, having no aggregation or sedimentation (Figure 8B).Effect of Chitosan Fractions with GA Additive on Stability of Ready to Drink (RTD) Black Tea-based Beverages

[0119] The Mw of chitosan, in combination with GA as an additive, significantly influenced the stability of the RTD black tea-based beverages. The OD of the control RTD black tea beverage was found to be 0.17, whereas the OD of the beverage with MMC and LMC was 0.12 and 0.13, respectively, after 35 days of storage (Figure 9). Incorporation of GA significantly improved the OD values of the beverage with - 23 -10369140LMC+Additive-2 (5% GA) and LMC+ Additive-3 (10% GA) displaying the OD of 0.18 and 0.19, respectively, which was higher than that of the control. Also, visually the beverage with LMC+Additive-2 (Figure 10D) looked more stable than MMC (Figure 10B) and LMC (Figure 10C) at the same dosage.Effect of Chitosan Fractions with GA Additive on Stability of Cold Coffee Drink-based Beverages

[0120] The Mw of chitosan, in combination with GA as an additive, significantly influenced the stability of the cold coffee drink-based beverages. For example, the OD of the control cold coffee beverage was found to be 1.07, whereas the OD of the beverage with LMC was 0.75, after 35 days of storage (Figure 11). Incorporation of GA significantly improved the OD values of the beverage; the OD of LMC+Additive-2 containing beverage was 0.84, which was higher than that of the LMC without GA additive.Antimicrobial Activity of Chitosan-based Formulations in Cold Coffee Drink-based Beverages

[0121] The LMC+Additive-2 formulation demonstrated antimicrobial activity against Zygosaccharomyces baili (Z. baili), Escherichia coli (E. coli) and Aspergillus niger (A. niger) at 50 ppm dosage in cold coffee drink-based beverages. The formulation completely inhibited the growth of Z. baili (Table 1), and reduced the count of E. coli (Table 2) and A. niger (Table 3) by 0.4 Log CFU / mL (Colony Forming Unit / g) after 21 days of storage.

[0122] Table 1. Antimicrobial activity of LMC chitosan-based formulation with additive-2 expressed as Log CFU / mL against Z. baili at 50 ppm dosage.- 24 -10369140Count (Log CFU / mL)Day 1 7 21Control 1 0 0LMC+Additive-2 o 0 0

[0123] Table 2. Antimicrobial activity of LMC chitosan-based formulation with additive-2 expressed as Log CFU / mL against E. coli at 50 ppm dosage.Count (Log CFU / mL)Day 1 7 21Control 2.65 2.33 1.88LMC+Additive-2 2 48 1 91 1 42

[0124] Table 3. Antimicrobial activity of LMC chitosan-based formulation with additive-2 expressed as Log CFU / mL against A. nigerai 50 ppm dosage.Count (Log CFU / mL)Day 1 7 21Control 2.47 2.27 2.16LMC+Additive-2 2 18 2 01 1 74Mold Inhibition Activity of Chitosan-based Formulations in Beverages

[0125] The orange juice, peach drink and cola-based beverages containing chitosan-based formulations were inoculated with A. niger to test the mold inhibition activity of the formulations. The LMC chitosan-based formulation at the dosage of 50 ppm doubled the shelf-life of the orange-juice based beverage as the control beverage was spoiled at day 8, whereas the beverage containing LMC chitosan-based formulation was spoiled at day 16 (Figures 12 and 13). Similarly the LMC chitosan- based formulation increased the shelf-life of the cola-based beverage by more than 5 - 25 -10369140days, as it remained spoilage free for 41 days (Figure 14). The LMC chitosan-based formulation also worked efficiently against P. chrysogenum in cola-based beverages. The LMC chitosan-based formulation at 50 ppm increased the shelf-life of the colabased beverage by more than 14 days (Figure 15). The MMC and MMC+Additive-6 (10% maltodextrin) worked very well in peach-drink based beverages as the chitosan formulations increased the shelf-life of the beverage by more than 35 days (Table 4). The control peach-drink beverage was spoiled within 22 days but the beverage sample containing MMC chitosan-based formulations was not spoiled after 60 days of storage. Similarly, the MMC and MMC+Additive-6 worked very well against P. chrysogenum in peach-drink based beverages as the chitosan formulations increased the shelf-life of the beverage by more than 35 days (Table 5). The control peach-drink beverage was spoiled within 22 days but the beverages with MMC was not spoiled up to 51 days and the MMC+Additive-6 chitosan-based formulations was not spoiled after 60 days of storage.

[0126] Table 4. Mold inhibition activity of MMC chitosan-based formulation against A. niger in Peach drink-based beverage

[0127] Table 5. Mold inhibition activity of MMC chitosan-based formulation against P. chrysogenum in Peach drink-based beverageEffect of Chitosan Fractions with GA Additive in the Streaming Potential (SP) of Orange Juice-based Beverages- 26 -10369140

[0128] The Mw of chitosan, in combination with GA as an additive, significantly influenced the SP values of the orange juice-based beverages (Figure 16). For example, the SP of the control orange juice beverages was measured to be -80 mV. Incorporation of 50 ppm of MMC chitosan-based formulation changed the colloidal balance of the beverage and shifted the total surface charge of the beverage to positive, which could be expected due to the cationic nature of chitosan polymer. However, incorporation of LMC chitosan-based formulation (at 50 ppm) maintained the surface charge as negative (-48 mV) and GA addition (in LMC) further increased the SP and stabilized the beverage; the same effect as discussed for the OD values previously (Figure 5). The effect of GA in the stabilization of the beverages was more pronounced at 100 ppm dosing of the chitosan-based formulations where the SP was higher for LMC with the GA additive.Effect of Chitosan Fractions with Maltodextrin (MD) Additive in the SP of Peach Drinkbased Beverages

[0129] The Mw of chitosan, in combination with MD as an additive, significantly influenced the SP values of the peach drink-based beverages (Figure 17). For example, the SP of the control peach beverages was measured to be -70 mV. Incorporation of chitosan-based formulations shifted the total surface charge of the beverage to positive, which could be expected due to the cationic nature of the chitosan polymer. However, based on the visual observation (Figure 8) and OD values (Figure 7) the beverages were stable, which could explain their high SP values (>125+ mV), with the incorporation of 100 ppm of MMC chitosan-based formulations and MMC with additives. From Figure 17, it can be seen that at 100 ppm dosage, the SP value of MMC chitosan-based formulations (+130 mV) was higher than that of the LMC (+78 mV), which could indicate increased colloidal stability of the beverages. While not wishing to be limited by theory, an increase in net surface charge can be interpreted as an increase in the repulsion of the ionic species present in the beverage, which prevents aggregation and sedimentation. The 200 pm of MMC chitosan-based formulations increased the SP, and MMC+Additive-4 (1 % maltodextrin) formulation had the highest SP for both the 100 and 200 ppm dosage.- 27 -10369140FULL CITATIONS FOR DOCUMENTS REFERRED TO IN THE APPLICATIONMourya, V.K.; Inamdar, N.N.; Choudhari, Y.M. Chitooligosaccharides: Synthesis, characterization and applications. Polym. Sci. Ser. A 2011 , 53, 583-612)Mohammed, M.A.; Syeda, J.T.M.; Wasan, K.M.; Wasan, E.K. An Overview of Chitosan Nanoparticles and Its Application in Non-Parenteral Drug Delivery. Pharmaceutics 2017, 9, 53)Chung, Y.-C., Yeh, J.-Y., & Tsai, C.-F. (2011). Antibacterial characteristics and activity of water-soluble chitosan derivatives prepared by the Maillard reaction. Molecules, 16(10), 8504-8514.Qin, C. Q., Du, Y. 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A Review on Chitin and Chitosan Polymers: Structure, Chemistry, Solubility, Derivatives, and Applications. ChemBioEng Rev 2015, 2, No. 3, 204-226Chen, J. S., Moschakis, T., & Pugnaloni, L. A. (2006). Surface topography of heat-set whey protein gels by confocal laser scanning microscopy. Food Hydrocolloids, 20(4), 468-474.Sabet, S., Rashidinejad, A., Melton, L. D., Zujovic, Z., Akbarinejad, A., Nieuwoudt, M., ... McGillivray, D. J. (2021). The interactions between the two negatively charged polysaccharides: Gum Arabic and alginate. Food Hydrocolloids, 112, 106343.El Abbadi, A., & Erni, P. (2022). Rheology and tribology of chitosan / Acacia gum complex coacervates. Soft Matter, 18(40), 7804-7813.Moschakis, T., Murray, B. S., & Biliaderis, C. G. (2010). Modifications in stability and structure of whey protein-coated o / w emulsions by interacting chitosan and gum arabic mixed dispersions. Food Hydrocolloids, 24(1), 8-17.Musa, H. H., Ahmed, A. A., & Musa, T. H. (2019). 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Claims

CLAIMS:1 . A food or beverage antimicrobial composition, comprising: chitosan having an average molecular weight (Mw) of about 2 to about 100 kDa; an organic acid; and an additive selected from gum arabic, maltodextrin, gellan gum, guar gum, xanthan gum, psyllium, polyglutamate, carrageenan, pectin, alginate, hyaluronic acid, dextran, chondroitin, polyphosphate, and combinations thereof.

2. The composition of claim 1 , wherein the additive is gum arabic.

3. The composition of claim 1 , wherein the additive is maltodextrin.

4. The composition of any one of claims 1 to 3, wherein the average Mw of the chitosan is about 2 kDa to about 50 kDa.

5. The composition of any one of claims 1 to 3, wherein the average Mw of the chitosan is about 50 kDa to about 100 kDa.

6. The composition of any one of claims 1 to 5, wherein the composition further comprises water and is a solution.

7. The composition of claim 6, wherein the chitosan is present in the composition in an amount of about 2 % w / v to about 10% w / v based on the total volume of the composition.

8. The composition of claim 7, wherein the chitosan is present in the composition in an amount of about 3 % w / v to about 8 % w / v based on the total volume of the composition.

9. The composition of claim 8, wherein the chitosan is present in the composition in an amount of about 5% w / v based on the total volume of the composition.- 31 -1036914010. The composition of any one of claims 1 to 9, wherein the chitosan has a degree of deacetylation that is greater than 50%.

11. The composition of claim 10, wherein the degree of deacetylation of the chitosan is greater than 80%.

12. The composition of any one of claims 1 to 11 , wherein the additive is present in the composition in an amount of about 0.5 % w / v to about 20 % w / v based on the total volume of the composition.

13. The composition of claim 12, wherein the additive is present in the composition in an amount of about 5 % w / v to about 10 % w / v based on the total volume of the composition.

14. The composition of any one of claims 1 to 13, wherein the organic acid is selected from citric, acetic, malic, formic, propionic and tartaric acids.

15. The composition of claim 14, wherein the organic acid is citric acid.

16. The composition of any one of claims 1 to 15, wherein the organic acid is present in the composition in an amount of about 1 % w / v to about 15 % w / v based on the total volume of the composition.

17. The composition of any one of claims 1 to 16, wherein the composition is in the form of a liquid solution or a solid.

18. The composition of any one of claims 1 to 17, wherein the composition is in the form of an aqueous solution.

19. A food or beverage product comprising the composition of any one of claims 1 to 18.

20. The food or beverage product of claim 19, wherein the composition is present in the food or beverage product in an amount of about 10 ppm to about 400 ppm.- 32 -1036914021 . The food or beverage product of claim 20, wherein the composition is present in the food or beverage product in an amount of about 25 ppm to about 200 ppm.

22. The food or beverage product of any one of claims 19 to 21 , wherein the composition substantially reduces or prevents formation of insoluble chitosan aggregates in the food or beverage product compared to formation of insoluble chitosan aggregates in the food or beverage product without the composition.

23. The food or beverage product of any one of claims 19 to 22, having a preservation time at room temperature that is increased by at least 10%, by at least 30% or by at least 50%, compared to a preservation time of the food or beverage product without the composition.

24. The food or beverage product of any one of claims 19 to 23, wherein the antimicrobial composition inhibits the growth of one or more microorganisms selected from bacteria, yeast and mold.

25. The food or beverage product of claim 24, wherein the microorganism is a gram-negative bacteria.

26. The food or beverage product of claim 25, wherein the gram-negative bacteria is Escherichia coli (E. coli).

27. The food or beverage product of claim 24, wherein the microorganism is a yeast.

28. The food or beverage product of claim 27, wherein the yeast is Zygosaccharomyces baili (Z. baili).

29. The food or beverage product of claim 24, wherein the microorganism is a mold.

30. The food or beverage product of claim 29, wherein the mold is Aspergillus niger (A. niger).

31. The food or beverage product of claim 29, wherein the mold is Penicillium chrysogenum (P. chrysogenum).- 33 -1036914032. The food or beverage product of any one of claims 19 to 31 , which is a beverage product.

33. The food or beverage product of claim 32, wherein the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juice-containing beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks.

34. The food or beverage product of any one of claims 32 to 33, wherein the beverage product further comprises one or more of glycerol, propylene glycol, and ethylenediaminetetraacetic acid.

35. The food or beverage product of any one of claims 32 to 34, wherein the composition reduces or prevents the turbidity of the beverage product.

36. A method of stabilizing and / or preserving a food or beverage product comprising adding an effective amount of one or more compositions of any one of claims 1 to 18 to the product.

37. The method of claim 36, wherein the food or beverage product is stabilized and / or preserved at room temperature by at least 10%, by at least 30% or by at least 50%, longer than a food or beverage product without the one or more compositions.

38. The method of claim 36 or 37, wherein the food or beverage product is a beverage product.

39. The method of claim 38, wherein the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juicecontaining beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks.

40. The method of any one of claims 36 to 39, wherein the effective amount of the one or more compositions is about 10 ppm to about 400 ppm.- 34 -1036914041. A method of inhibiting growth of one or more microorganisms in a food or beverage product comprising adding an effective amount of one or more compositions of any one of claims 1 to 18 to the product.

42. The method of claim 41 , wherein the one or more microorganisms are selected from bacteria, yeast and mold.

43. The method of claim 42, wherein the microorganism is a gram-negative bacteria.

44. The method of claim 43, wherein the gram-negative bacteria is Escherichia coli (E. coli).

45. The method of claim 42, wherein the microorganism is a yeast.

46. The method of claim 45, wherein the yeast is Zygosaccharomyces baili (Z. baili).

47. The method of claim 42, wherein the microorganism is a mold.

48. The method of claim 47, wherein the mold is Aspergillus niger (A. niger).

49. The method of claim 47, wherein the mold is Penicillium chrysogenum (P. chrysogenum).

50. The method of any one of claims 41 to 49, wherein the food or beverage product is a beverage product.

51. The method of claim 50, wherein the beverage product is selected from cola, cola-containing beverages, soda, soda-containing beverages, juice, juicecontaining beverages, coffee, coffee-containing beverages, kombucha, flavored water, tea, tea-containing beverages and fermented drinks.

52. The method of any one of claims 41 to 51 , wherein the effective amount of the one or more compositions is about 10 ppm to about 400 ppm.- 35 -10369140

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