MXPA06002068A - Encapsulated antimicrobial material. - Google Patents
Encapsulated antimicrobial material.Info
- Publication number
- MXPA06002068A MXPA06002068A MXPA06002068A MXPA06002068A MXPA06002068A MX PA06002068 A MXPA06002068 A MX PA06002068A MX PA06002068 A MXPA06002068 A MX PA06002068A MX PA06002068 A MXPA06002068 A MX PA06002068A MX PA06002068 A MXPA06002068 A MX PA06002068A
- Authority
- MX
- Mexico
- Prior art keywords
- antimicrobial material
- encapsulated
- food
- further characterized
- nisin
- Prior art date
Links
- 239000000463 material Substances 0.000 title claims abstract description 682
- 230000000845 anti-microbial effect Effects 0.000 title claims abstract description 540
- 238000000034 method Methods 0.000 claims abstract description 140
- 239000004599 antimicrobial Substances 0.000 claims abstract description 59
- 230000008569 process Effects 0.000 claims abstract description 43
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Classifications
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- A—HUMAN NECESSITIES
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- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
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- A—HUMAN NECESSITIES
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- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/12—Preserving with acids; Acid fermentation
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
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- A—HUMAN NECESSITIES
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- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
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Abstract
The present invention provides an antimicrobial material in an encapsulated form, comprising (i) a core comprising an antimicrobial material and (ii) a shell of encapsulating material, wherein the shell of encapsulating material is impermeable to the antimicrobial material. The invention further provides a process for introducing an antimicrobial material into a foodstuff comprising (i) providing the antimicrobial material in an encapsulated form comprising a core of antimicrobial material and shell of encapsulating material, and (ii) introducing encapsulated antimicrobial material into or onto the foodstuff.
Description
ANTIMICROBIAL MATERIAL ENCAPSULATED FIELD OF THE INVENTION
The present invention relates to a method for introducing antimicrobial material into a food. The present invention also relates to an antimicrobial material.
BACKGROUND OF THE INVENTION
Antimicrobial materials are well known in the art. A well-known antimicrobial material is natamycin. Natamycin is a natural antifungal agent of polyene macrolide produced by the fermentation of the bacterium Streptomyces natalensis. Natamycin (formerly known as pimaricin) has an extry effective and selective mode of action against a very broad spectrum of yeasts and molds of food rot common with most strains being inhibited by concentrations of 1-15 ppm natamycin. Natamycin is accepted as a food preservative and used worldwide, particularly for surface treatment of dry fermented cheese and sausages. It has several advantages as a food preservative, including a broad spectrum of activity, efficacy at low concentrations, lack of resistance and activity over a wide range of pH. The natural aqueous suspensions of natamycin are very stable, but natamycin has poor stability in acid or alkaline conditions, in the presence of light, oxidants and heavy metals. For example, natamycin can be used in pasteurized fruit juice to prevent putrefaction by heat-resistant molds such as Byssochlamys. The pH of the juice, however, promotes the degradation of natamycin during pasteurization as well as during storage if the juice is not refrigerated. Natamycin is also degraded by heat processing at a high temperature, such as occurs when baking confectionery products in an oven. At extrpH conditions, such as pH less than 4 and greater than 10, natamycin is rapidly inactivated with formation of various types of decomposition products. The acid hydrolysis of natamycin releases the inactive amino sugar, mycosamine. Additional degradation reactions result in the formation of dimers with a triene more than a tetrane group. Heating to a low pH can also result in the decarboxylation of the aglycone. The alkaline hydrolysis results in saponification of the lactone. The acid degradation products (aponatamycin, the aglycone dimer and mycosamine), and the alkaline or UV degradation products proved to be even safer than natamycin in toxicology tests, but are biologically inactive. Natamycin is usually dosed into or on a food as a powder or as a suspension of an aqueous natamycin. This type of dosage form leaves the natamycin unprotected under the conditions of processing and use. Natamycin powder, although mixed with excipients such as lactose, can also be sticky to handle and can cause dust problems within food processing plants. In addition, natamycin is as highly effective as an antifungal compound that can adversely affect the processing of products intended to be preserved if this depends on the activity of the desired fungus. Therefore, there is a need for a protected dose form of natamycin. A general description of natamycin and its current uses can be found in Thomas, L.V. and Delves-Broughton, J. 2003. Natamycin. In: Encyclopedia of Food Sciences and Nutrition. Eds. B. Caballero, L. Trugo and P. Fingías, pp. 4109-4115. Elsevier Science Ltd. Bacteriocins are antimicrobial proteins or peptides that can be produced by certain bacteria, which can kill or inhibit the growth of closely related bacteria. Bacteriocins produced by lactic acid bacteria are of particular importance since they have great potential for food preservation and for the control of food pathogens (Wessels et al., 1998). The best known bacteriocin is nisin, which is the only bacteriocin currently authorized as an additive for food. Nisin is produced by the fermentation of the bacterium from the Lactococcus lactis subsp. Lactis, and sold as the commercial extract Nisaplin® Natural Antimicrobial (Danisco). Nisin has an unusually broad antimicrobial spectrum for a bacteriocin, being active against most gram-positive bacteria (e.g., species of Bacillus, Clostridium, Listeria, lactic acid bacteria). It is usually not effective against gram-negative bacteria, yeasts or molds. Nisin is allowed as a food conservator worldwide but its levels of use and approved food applications are strictly regulated, varying from one country to another. Other bactereocins have since been discovered with potential as food preservatives, e.g., pediocin, lacticin, succinate, lactococcin, enterococin, plantaricin, leucocin. They are also active, although usually with a narrower spectrum, against gram-positive bacteria. Its nutritional use is currently restricted to the production of the bacteriocin in situ, that is, making the grower grow in the food. Food safety and the prevention of food putrefaction is a current global concern, particularly with an increasing tendency for convenience foods such as ready-to-eat meals, soups, sauces or snacks. Food rot is an economic problem for the food manufacturer. Food manufacturers need to protect the health and safety of the public by providing products that are safe when eaten. These foods must have a shelf life guaranteed either at storage at a cooled or ambient temperature. Consumers prefer foods with good taste of high quality - this is difficult to achieve with chemical preservatives, inhospitable heating regimes and other processing measurements. Food safety and protection is best achieved with a multiple preservation system using a combined approach of lighter processing and natural preservatives. Microorganisms in food are also less able to adapt and grow in preserved food with different conservation measures. There is a lot of concern about the safety of food and the growth of foodborne pathogens such as Listeria monocytogenes. This particular pathogen can grow at low temperatures, which are often used as a measure of additional preservative. Food pathogens can sometimes be adapted to different preservatives and storage conditions, so a combination of conservation measures can be more successful than individual measures. Cooked meat cuts are new generation convenience products, which are now offered to consumers. The preparation of these cuts of meat generally involve the injection or stirring of the raw meat in polyphosphate brine to increase the softness, moistness and volume of the meat. The meat is then cooked before being distributed to the retail establishments and their subsequent purchase and consumption by the consumer. Most of the processing for these meats now involves the "internal cooking" system in which the meat is cooked in plastic bags or plastic film. Whole cuts can be deboned, pumped with polyphosphate brine and scrambled or handled for a short period. This decreases the brine evenly and also achieves a layer of exudate on the surface that helps the plastic package to adhere tightly to the surface of the meat. Large cuts are usually packed with gas or vacuum in plastic bags. These cooked meat products are often considered to be of good quality and healthy, since they can be low in fat with a minimum salt content. They may not necessarily be reheated by the buyer for consumption. These minimally processed products are based on refrigeration to ensure the stability and safety of cooked meat during shelf life, which can be as long as 90 days (Vamam and Sutherland, 1995). Putrefaction of cooked meats, if post-processing contamination is a factor, would be due to the Bacillus and Clostndium gram-positive heat-resistant bacteria, particularly if the meat is exposed to temperatures above 7 ° C. Putrefaction due to these organisms can be rapid if the meat is exposed to temperatures as high as 15 ° C or higher. If the meat has not been cooked enough, heat-resistant species of Entérococcus or Lactobacillus can survive, many of which can grow at refrigeration temperatures. If the product has been handled after cooking and repacked and packed under vacuum, putrefaction is often associated with Lactobacillus, Leuconostoc or Camobacterium. Brochothríx thermosphacta, another Gram positive bacterium, can also cause problems. Gram-negative bacteria will only be a problem in cooked unpackaged meats, or those that are packaged in air permeable film. Molds can develop on cooked meat cuts that have been exposed to air and whose surfaces have dried. There is also concern about post-processing contamination and growth of Listeria monocytogenes, a food pathogen that can grow at refrigeration temperatures. It would be a benefit to both the public in terms of safety and to manufacturers in terms of economy and reputation, if an effective conservative could somehow be applied to the surface layer of cooked meat. Raw whole muscle meat is also increasingly sold as a chilled convenience meat product that is easily prepared and smoothed for the consumer to cook. The meat is usually covered with a marinade and then packed under vacuum in a transparent bag. The marinade can be applied and simply wet on the surface of the meat, or the meat can be stirred in the marinade to increase its softening and penetration effect. This fresh marinated meat, packed under vacuum can be maintained for up to 28 days at refrigeration temperatures before being purchased by the consumer and subsequent cooking at home. These meat products are considered fresh value-added meats and cover a wide range of raw meats (pork, chicken, beef, ground beef, steaks, chopped meats, cuts, etc.). The combination of the acidic nature of marinade and the lack of oxygen in bags packed under vacuum means that gram-positive lactic acid bacteria are associated with putrefaction of their products (Susiluoto et al., 2003). Nisin is a natural preservative that has been used safely in food for almost 50 years. It is effective against gram-positive bacteria including lactic acid bacteria, Brochothrix thermosphacta, Listeria monocytogenes, Bacillus and Clostridium. As the putrefaction associated with the meat products described above, is generally caused by gram-positive bacteria, nisin could be considered as part of a conservative system to guarantee or prolong the shelf life. However, the environment of both meat products is not favorable for the stability or activity of nisin. The brine and polyphosphate solutions used to inject raw meat are usually at an alkaline pH. The stability of nisin is optimal at pH 3 (Davies et al., 1998). The cooking process, particularly at high or neutral pH conditions, would lead to significant nisin degradation. In raw meat, nisin is vulnerable to degradation by proteases. A more specific concern is the inactivation of raw nisin in the formation of an adduct with glutathione in an enzyme-mediated reaction (Rose et al., 1999)., 2002, 2003). Numerous teachings of the prior art describe potential uses of nisin in foods. Examples are: • Caserío et al. (1979) describes the research on the use of nisin in meat products, cooked cured. Mortadella, Wurstel sausage, prosciutto. Target organisms: Staphybcoccus, sulfate-reducing clostridia. The prosciutto has been injected with nisin with brine then dissolved in diluted lactic acid. • Gola (1962) incorporated nisin in gelatin for canning of large hams. In the first experiment, the brines by injection were acidified to facilitate the solubility of the nisin. • Taylor &; Somers (1985) evaluate the antibotulinal effectiveness of nisin in bacon. Nisin was included in the brine formulation injected into the pig's belly. • Usborne et al. (1986) describes sensorial evaluation of bacon treated with nisin. Nisin was added to the brine pumping solution before being injected into the bacon. • US 2003/0108648 relates to compositions having bacteriostatic and bactericidal activity against spores of bacteria and vegetative cells and method for treating foods with them. »US 6207210 relates to a broad-spectrum antibacterial composition and method of application to food surfaces. • EP0770336 describes the injection of meat / brine cut solution in which a starter culture has produced a bacteriocin. · The article found at http://www.nal.usda.gov/fsrio/ppd/ars010f.htm about the work in the Research Unit, MARC mentions a presentation about "the antibacterial properties of marinated injectable beef". • WO2003 / 11058 relates to food preservation formulation comprising compound (s) derived from natural sources. Natural compounds are formulated and applied to a food and irradiation at < 3kGy results in decreased microflora compared to non-irradiated controls. Nisin is a preferred compound. · US 2003/0108648 teaches nisin as part of a combination for marinates. The extensive prior art does not address or solve the protection problems of antimicrobial materials such as nisin from environments, such as those in meat products, which are not favorable for the stability or activity of the antimicrobial material. The present invention solves the problems of the prior art. In one aspect, the present invention provides an antimicrobial material in an encapsulated form, comprising a core of antimicrobial material and shell of encapsulating material, wherein the mantle of the encapsulant material is impermeable to the antimicrobial material and optionally is physiologically acceptable. The term "encapsulation" is well known in the art. Encapsulation can be defined as the technology of packing a substrate (solids, liquids, gases) into another material. In the encapsulation, the material that has been trapped is referred to as the core material or the internal phase while the encapsulating material is referred to as the coating, the shell material or the vehicle. Encapsulating materials are also commonly referred to as core / shell materials. In one aspect, the present invention provides a method for producing an antimicrobial material in an encapsulated form, comprising co-processing an antimicrobial material with an encapsulating material, to cause the material to encase the antimicrobial material within a shell, and recovering the antimicrobial material, wherein the shell of the encapsulating material is impermeable to the antimicrobial material and optionally is physiologically acceptable. In one aspect, the present invention provides a method for introducing an antimicrobial material into a food comprising (i) providing the antimicrobial material in an encapsulated form comprising a core of the antimicrobial material and a shell of encapsulating material, (i) introducing antimicrobial material encapsulated within or on the food, preferably by (a) injecting the antimicrobial material encapsulated in the food or (b) by stirring the antimicrobial material encapsulated with the food. In one aspect, the present invention provides a food prepared by a process for introducing an antimicrobial material into a food comprising (i) providing the antimicrobial material in an encapsulated form comprising a core of antimicrobial material and shell of encapsulating material, (ii) ) introduce encapsulated antimicrobial material in or on food, preferably by (a) injecting the antimicrobial material encapsulated in the feed or (b) by stirring the antimicrobial material encapsulated with the feed. In one aspect, the present invention provides a food obtainable by a process for introducing an antimicrobial material into a food comprising (i) providing the antimicrobial material in an encapsulated form comprising a core of antimicrobial material and shell of encapsulating material, (ii) ) introducing encapsulated antimicrobial material into or on the food, preferably by (a) injecting the antimicrobial material encapsulated in the food or (b) stirring the antimicrobial material encapsulated with the food. The aspects of the invention are defined in the appended claims. The inventors of the present have found that by providing the antimicrobial materials in an encapsulated form, the materials can be protected against environments, such as those in meat products, which are not favorable for the stability or activity of the antimicrobial material. However, by injecting the encapsulating antimicrobial material into the feed or by stirring the antimicrobial material encapsulated with the feed, the encapsulated antimicrobial material can be effectively introduced or onto the feed. The inventors of the present have found the injection particularly advantageous and surprisingly. The prior art methods have directly injected non-encapsulated antimicrobial materials such as nisin into food products. Now, the inventors of the present have found that a "shell" can be provided on the antimicrobial material which is able to withstand the demanding physical forces exerted on the antimicrobial material encapsulated during the injection. In particular, the injection exerts high pressures and produces stress on the material to be injected. Now, the inventors of the present have found that a "shell" can be provided on e! antimicrobial material that is able to protect the antimicrobial material against adverse conditions and / or allows sustained / controlled release. The present invention provides a method for delivering an antimicrobial material and an antimicrobial material per se that is resistant to unwanted degradation and that can be released to provide a long-term antimicrobial effect. For ease of reference, these and other aspects of the present invention are now described under appropriate section headers. However, the teachings under each section are not necessarily limited to each particular section.
Favorite aspects
Antimicrobial Materials In a preferred aspect, the antimicrobial material is an antibacterial material. In a preferred aspect, the antimicrobial material is a bacteriocin. The antimicrobial material, such as a bacteriocin, can typically be selected from materials (bacteriocins) that can be used as preservatives in foods. Preferably, the antimicrobial material is selected from bacteriocins containing lanthionine, bacteriocins derived from Lactococcus, bacteriocins derived from Streptococcus, bacteriocins derived from Pediococcus, bacteriocins derived from Lactobacillus, bacteriocins derived from Camobacterium, bacteriocins derived from Leuconostoc, bacteriocins derived from Enterococcus and mixtures of the same. Preferably, the antimicrobial material is at least nisin. Preferably, the antimicrobial material consists of nisin. Nicin is a bacteriocin containing lanthionine (US 5691301) derived from Lactococcus lactis subsp. Lactis (formerly known as Streptococcus-lactis) (US 5573801). In a preferred aspect of the present invention the bacteriocin used in the present invention is at least nisin. As described in US 5573801, nisin is a polypeptide bacteriocin produced by the lactic acid bacteria, Lactococcus lactis subsp. Lactis (formerly known as Streptococcus lactis group N). Nisin is a collective name that represents several closely related substances that have been designated as nisin compounds A, B, C, D and E (De Vuyst, L. and Vandamme, EJ 1994. Nisin, a lantibiotic produced by Lactococcus lactis subsp. lactis: properties, biosynthesis, fermentation and applications In: Bacteriocins of lactic acid bacteria Microbiology, Genetics and Applications Eds .: De Vuyst and Vandamme, Blackie Academic and Professional, London). The structure in nisin properties are also described in the article by E. Lipinska, entitled "Nisin and Its Applications", The 25th Proceedings of the Easter School in Agriculture Science at the University of Nottingham, 1976, pp. 103-130 (1977), said article is incorporated herein by reference. In 1969, the FAO / WHO Joint Expert Committee on Food Additives established specifications for the purity and identity of nisin (FAO / WHO Joint Expert Committee on Food Additives, 1969. Specifications for the quality and purity of some antibiotics. Report Series No. 430). This committee recognized nisin as a safe and legal conservator based on extensive toxicological testing. Nisin has the food additive number E234 and is classified as GRAS (Generally Recognized as Safe) (Food and Drug Administration, 1988. Nisin preparation: Affirmation of GRAS status as a direct human ngredient, Federal Regulations 53: 11247). The international activity unit (hereinafter IU) was identified as 0.001 mg of a reference preparation of nisin international. Nisaplin® Natural Antimicrobial is the brand name for a nisin concentrate containing 1 million IU per gram, which is commercially available from Danisco. Nisina is a recognized and accepted food preservative with a long history of safe and effective use in food. There are several reviews of nisin, eg, Hurst 1981; 1983; Delves-Broughton, 1990; De Vuyst and Vandamme, 1994; Thomas et al. 2000; Thomas & Delves-Broughton, 2001). Nisin was discovered about 50 years ago and the first commercial preparation, made in 1953, was Nisaplin®. Nisin has several characteristics that make it particularly suitable as a food preservative. He has gone through extensive toxicological tests to prove his safety. It is stable to heat, stable to acid and effective against a broad spectrum of gram-positive bacteria. It is usually not effective against gram-negative bacteria, yeasts or molds but activity against gram-negative bacteria and yeast has been reported in the presence of chelating agents (PCT / US 8902625. WO 89/12399). Nisin is an effective preservative in pasteurized and heat-treated foods (v.gr., processed cheese, cheese, pasteurized milk, dairy desserts, cream, mascarpone and other dairy products, puddings such as semolina, tapioca etc., pasteurized liquid egg, pasteurized potato products, soy products, crumpets, hard buns (pikelets), dough cakes (flapjacks), processed meat products, beverages, soups, sauces, ready-to-eat meals, canned foods, vegetable beverages) and low acid foods such as salad dressings, sauces, mayonnaise, beer, wine and other drinks. Although some loss of activity can be expected when used with processed foods, this can be improved eg by increasing the amount of nisin applied. Effective nisin levels for preserving food are reported to vary from 25-500 lU / g or more. Other effective levels would be appreciated by one skilled in the art. For example, levels of 50-500 lU / g can be used. Since the discovery of the first bacteriocin, nisin, many other bacteriocins have been found (Hoover, 1993; Ray & Daeschel, 1994; Axelsen, 1998; Naidu, 2000; Ray et al. 201; Ray & Miller, 2003). The pediocin bacteriocin, produced by Pediococcus pentosaceus, P. acidilactici, or Lactobacillus plantarum, can be used in the present invention. Like nisin, different pediocin structures have been described. At present, pediocin and other bacteriocins are not allowed as food additives but their antibacterial activity can be achieved through the production of the bacteriocin in situ, as a consequence of the growth of the producer organism in the food. This is the purpose of commercial protective crops such as HOLDBAC ™ Listeria (Danisco). Pediocin has a narrower antimicrobial spectrum compared to nisin, but there is much interest in its food safety capacity to kill, prevent or control the growth of the food pathogen of Listeria monocytogenes (Ray &Miller, 2000). Other bacteriocins can be used in the present invention, including those generally referred to as Divercina, leucocin, mesentericin, succinin, curvacin, bavaricin, acidocin, bifidocin, carnobacteriocin, pisicocin, piscicolin, mundticin, enterocin, thermophilin, lacticin, plantarcin, lactococin, divercin , diplococin, mesenterocin, leukonosin, carnosine, acidophilin, lactacin, brevicin, lactocin, helevticin, reutericin, propionicin. Preferably, the antimicrobial material is at least natamycin. Preferably, the antimicrobial material consists of natamycin.
Microorganisms As described herein, the present invention can prevent and / or inhibit the growth of and / or kill a microorganism in a material. This can slow or stop a microorganism such as bacteria, or kill a microorganism present in contact with the present composition. In one aspect, the antimicrobial material is present in an amount to provide microbicidal or microbiostatic effect. In one aspect, the bacteriocin and the extract are present in an amount to provide a microbicidal or microbiostatic effect. In a highly preferred aspect, the microbicidal or microbiostatic effect is a fungicide or fungistatic effect, which optionally includes an effect against yeast. In a preferred aspect, the fungicidal or fungistatic effect is with respect to an organism selected from fungi or yeast associated with putrefaction of food or food-borne diseases. In a preferred aspect the fungicidal or fungistatic effect is with respect to an organism selected from Yeasts: Candida species (e.g., C. krusei, C. parapsilosis, C. utilis, C. valida), Dekkera (v. ., D. bruxellensis), Debaryomyces (e.g., D. hansenii), Hanseniaspora (e.g., H. uvarum) Kluyveromyces (e.g., K. loctis), Pichia (P. membranaefaciens), Rhodosporium , Rhodotorula. { Rh mucilaginous), Saccharomyces (e.g., S. bayanus, S. boulardi, S. carlsbergensis, S. cerevisiae, S. exiguus, S. florentinus, S. unisporus), Zygosaccharonmyces (e.g., Z. rouxli , Z baili). Molds: Aspergillus species (eg, A. niger, A. restricctus, A. versicolor), Byssochlamys (eg, B. fulva, B. nivea), Eupenicillium, Eurotium, Fusarium, Geotrichum, Mucor, Neosañorya (e.g., N. fischeri var. Fischeri), Penicillium (e.g., P. aurantiogriseum, P. brevicompactum, P. camembertii, P. candidum, P. chrysogenum, P. commune, P. corylophilum, P. cyclopium, P. discolor, P. nalg / ovense, P. rogueforti), Talaryomyces (e.g., T. macrosporus). In a highly preferred aspect, the microbicidal or microbiostatic effect is a bactericidal or bacteriostatic effect. It is advantageous if the bactericidal or bacteriostatic effect is effective with respect to gram-positive bacteria and gram-negative bacteria. Preferably, the bactericidal or bacteriostatic effect is with respect to gram-positive bacteria. In a preferred aspect, the bactericidal or bacteriostatic effect is with respect to an organism selected from gram-positive bacteria associated with food spoilage or food-borne disease including species of Bacillus, Bacillus subtilis, Bacillus cereus, Listeria species, Listeria monocytogenes, lactic acid bacteria, lactic acid putrefaction bacteria, Lactobacillus species, Staphylococcus aureus species, Clostridium, C. sporogenes, C. tyrobutyrícum and C. botulinum (when the antimicrobial material is recognized as effective against C. botulinum or is part of an effective system against C. botullinum). In a preferred aspect, the bactericidal or bacteriostatic effect of the invention in combination with a chelating agent is with respect to an organism selected from other microorganisms associated with food spoilage or food-borne disease, including yeasts, molds and gram-negative bacteria. Including Escherichia coli, Salmonella species, and Pseudomonas species. In a preferred aspect, the bactericidal or bacteriostatic effect is with respect to lactic acid bacteria such as Lactobacillus, Leuconostoc, Camobacterium, and Enterococcus; Listeria monocytogenes, heat-resistant spore-forming bacteria such as Bacillus and Clostridium; and Brochothríx thermosphacta. In a preferred aspect, the bactericidal or bacteriostatic effect is with respect to Bacillus cereus. In a preferred aspect, the bactericidal or bacteriostatic effect is with respect to Listeria monocytogenes.
Encapsulated antimicrobial material Encapsulation technology has been applied to a number of food ingredients, usually to cover the taste or activity. The present invention is based on the realization that the unexpected benefits are derivable from the encapsulation of antimicrobial materials. Koontz & Marcey, 2003, J Agrie Food Chemistry 51: 7106-71 describes the formation of an inclusion product of natamycin / cyclodextrin.
Cyclodextrin acts as host molecules to protect mainly against light, but also low pH, heat and oxidation. However, this natamycin / cyclodextrin complex is not a true encapsulation. A natamycin molecule will not "fit" gamma-cyclodextrin, therefore it can only be considered a partial encapsulation. Acid conditions tend to destabilize this type of complex, releasing the content of the cyclodextrin molecule and the natamycin molecule is not completely enclosed and protected by the cyclodextrin molecules. Koontz et al. 2003. J Agrie Food Chemistry 51: 7111-7114 has also described the stability of natamycin and its cyclodextrin inclusion complexes in aqueous solution. EP115618 discloses an antifungal anti-cake ingredient food ingredient wherein the anti-cake-forming agent is encapsulated and then treated with natamycin to provide antifungal activity. US 5,445,949 describes a process for recovering natamycin by separating a hydrophobic fermentation product such as natamycin. The method involves a step that includes encapsulation of a protein but no encapsulation of natamycin is mentioned. EP-A1-1382261 discloses the use of microbial inhibitors such as natamycin for confectionery products, including stable shelf-stable equipment for making snacks or meals. The microbial inhibitor is not protected by encapsulation. The patent application of E.UA No. 10 / 765,210, filed on January 28, 2004, refers to the protection of confectionery products by spraying the surface of the products with a suspension of natamycin and therefore to increase the life of products shelf. WO 89/033208 describes a polyene macrolide powder for liposome preparation. The polyene macrolide is encapsulated in liposomes to modify the pharmacokinetics of antifungal in systemic diseases. The liposome is designed for pharmaceutical use only. US 5,821,233 relates to an antifungal composition wherein natamycin is incorporated in porous silica to provide delayed release of natamycin in an aqueous medium. General descriptions of encapsulation procedures can be found in Shahidi, F., and X.- Q., Han. 1993. Encapsulation of food ingredient. Critical Reviews in Food Science and Nutrition 33 (6): 501-547. The encapsulation of mold inhibitors is described in Ranum, P., 1999. Encapsulated mold inhibitors-the greatest thing since sliced bread in Cereal Foods World, Vol 44, No. 5, p. 370-371. US 5,204,029 describes a process for preparing edible microcapsules containing a multiplicity of liquid nuclei. In the process, a water-in-oil emulsion, with the active ingredient dissolved in an internal aqueous phase, is spray-cooled, which causes solidification of the fat phase and entrapment of the aqueous phase as tiny droplets dispersed in a microcapsule. This procedure, however, leads to highly unstable microcapsules from which the water phase migrates from the inner part of the microcapsule to the external part. This also results in the condensation of water on the wall of the container. In Kirk-Othmer Encyclopedia of Chemical Technology, 3a. Ed. Vol. 15, pp. 473 to 474, a method is described in which the liquids encapsulated using a rotating extrusion head containing concentric nozzles. The process is only suitable for liquids or suspensions, and the products of the processes are large spheres having fusible coatings, such as fats or waxes. However, microcapsules that contain a single drop of liquid as a core are very susceptible to breakage. In his article, "Mass preparation and characterization of alginate microspheres" n Process Biochemistry 35 (2000) 885 to 888 Mofidi, N. et al., Describe a method for mass preparation of microspheres, in said method a sterilized alginate solution is prepared and the solution is then emptied into a reactor containing a non-aqueous phase, while being stirred. An alginate microdroplet emulsion is formed and an appropriate amount of crosslinker is added. The microspherical gel-alginate particles fall to the bottom and are collected by filtration. Similarly, Wong, T.W. et al., in J. Microencapsulation, 2002 Vol. 19, no. 4,511 to 522, describe characteristics of release of pectin microspheres and the method for preparing these microspheres. In this method, the pectin microspheres are prepared by a water-in-oil emulsion technique, in which minute droplets of pectin containing an active ingredient dispersed in a liquid hydrophobic continuous phase are hardened and collected by filtration. Microencapsulation by a coacervation-phase separation process is known from an article by Joseph A. Bakan in Controlled Relay Technologies, 1980 by Agis F. Kydonieus. The procedure consists of a series of three steps carried out under continuous agitation: (1) formation of three immiscible chemical phases; (2) coating deposition; and (3) coating stiffening. Sanghvi, S.P. and Nairn J.G. have studied the effect of the viscosity and interfacial tension of the particle size of cellulose acetate trimellitate microspheres. The results are presented in their article in J. Microencapsulation, 1992, Vol. 9, not 2,215 to 227. In their article in Lebensm. -Wiss. U. -Technol., 33, 80 to 88 (2000) Lee, S.J. and Rosenberg, M., describe a double emulsification and heat-gelling process for preparing microcapsules based on whey protein. The microcapsules prepared according to the method described are microcapsules based on whey protein containing an apolar core material. In their article in Science Vol. 298, November 1, 2002, Dinsmore et al. Describe selectively permeable capsules composed of colloidal particles. The capsules are manufactured by the self-assembly of colloidal particles on the bottom of emulsion droplets. After the particles are locked together to form elastic breastplates, the emulsion drops are transferred to a fresh continuous phase fluid that is the same as that of the inside of the drops. In a preferred aspect, the encapsulated antimicrobial material is a particulate form. The particle size can be important either in the injection aspect of the present invention or the stirring aspect. The choice of particle size, for example to a particular maximum average particle size, can assist in the introduction of the antimicrobial material encapsulated within or on the food. The inventors of the present invention have found that the injection aspect of the particle size is particularly important. The particle size, and in particular the maximum particle size, can determine the likelihood that the shell of the encapsulated antimicrobial material will withstand an injection procedure. In a preferred aspect, the encapsulated antimicrobial material has an average particle size of less than 500 μpp, preferably less than 300 μ? T ?, preferably less than 250 μ? , preferably less than 150 μ ??, preferably 50 to 150 μ ??. In some aspects, the encapsulated antimicrobial material may have an average particle size of less than 100 μ ??, or less than 50 μ ??, or less than 25 μ ??. As described above, one purpose of the present invention is to provide for the introduction of the antimicrobial material into or on the food in a form protected from degradation or inactivation. However, the antimicrobial material must of course be released when required to provide the antimicrobial effect that is its purpose. Therefore in a preferred aspect, the shell will be selected to provide sustained release of the antimicrobial material from the encapsulated animicrobial material. In one aspect of the present invention, the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material or to provide release under predetermined conditions. Suitable predetermined temperature conditions are: more than 50 ° C, preferably more than 60 ° C, preferably more than 70 ° C, preferably more than 72 ° C, preferably more than 75 ° C, preferably 72 to 78 ° C. In one aspect of the present invention, the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material. Preferably, the degeneration to be prevented is by one or more factors selected from thermal degradation, pH-induced degradation (either by acidic or alkaline pH), degradation by oxidation, degradation by light, degradation by protease and formation of adduct by glutathione
Therefore, in a further aspect, the present invention provides the use of an encapsulating material for the prevention, reduction or inhibition of the degeneration or inactivation of an antimicrobial material. Preferably, the degeneration to be prevented is by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation. The shell is or comprises or may be formed of any suitable material. The shell material useful in the invention may be selected from the group comprising fats, oils, waxes, resins, emulsifiers or mixtures thereof, which are preferably food grade. Preferably, the lacquer material is selected from the group comprising oils and fats of animal origin, oils of completely hydrogenated vegetable or animal origin, partially hydrogenated vegetable or animal oils, unsaturated, hydrogenated or fully hydrogenated fatty acids, monoglycerides and diglycerides of unsaturated, partially hydrogenated or fully hydrogenated fatty acids, partially hydrogenated or fully hydrogenated esterified fatty acids of monoglycerides and diglycerides, unsaturated, partially hydrogenated or fully hydrogenated free fatty acids, other emulsifiers, animal waxes, vegetable waxes, waxes of origin mineral, synthetic waxes, natural and synthetic resins and mixtures thereof. Oils and fats of animal origin are such as, but not restricted to, beef bait, mutton bait, lamb bait, lard or pork fat, sperm oil. Hydrogenated or partially hydrogenated vegetable oils such as, but not restricted to, canola oil, cottonseed oil, peanut oil, corn oil, olive oil, soybean oil, sunflower oil, safflower oil, oil of coconut, palm oil, linseed oil, stick oil and castor oil. Free fatty acids are such as, but not restricted to, stearic acid, palmitic acid and oleic acid. Other emulsifiers are such as, but not restricted to, polyglycerol esters, sorbitan esters of fatty acids. Waxes of animal origin are such as, but not restricted to beeswax, lanolin, shell wax or Chinese insect wax. Waxes of vegetable origin are such as but not restricted to carnauba, candelilla, bay berry or sugarcane waxes. Waxes of mineral origin are such as but not restricted to paraffin, microcrystalline oil, ozokerite, ceresin or montana. Synthetic waxes are such as but not restricted to low molecular weight polyolefin, ether-polyol esters and synthetic waxes from the Fisher-Tropsch process. Natural resins are such as rosin, balsam, lacquer and zein. The hydrocolloid shell material of the invention may be any food grade hydrocolloid that is capable of providing encapsulation, for example by the methods of the invention. The material can be selected from the group comprising hydrocolloids, sodium alginate, gum arabic, gellan gum, starch, modified starch, guar gum, agar gum, pectin, amidified pectin, caragenan, xanthan, gelatin, chitosan, mesquite gum , hyaluronic acid, cellulose derivatives such as cellulose acetate phthalate, hydroxypropylmethyl cellulose (HP C), methyl cellulose, ethyl cellulose and carboxy methyl cellulose (CMC), acrylic methyl copolymers, such as Eudragit®, Psyllium, tamarind, xanthan, locust bean gum, whey protein, soy protein, sodium caseinate, food grade protein, shellac, zein, any synthetic or natural water soluble polymers, and mixtures thereof. Additional preferred lacquer materials are selected from fats, emulsifiers, waxes (of animal, vegetable, mineral or synthetic origin), liposome-forming lipids (such as glycerophospholipids and sterols), hydrocolloids, natural or synthetic polymers and mixtures thereof. Preferred materials are materials that are insoluble in brine or can be made insoluble in brine by entanglement, compression or other means. Preferably, the glycerophospholipids are selected from phosphatidyl, phosphatidiethanolamines and phosphatidylglycerols. Preferably, the sterols are selected from cholesterol, ergosterol, lanosterol and stigmasterol. Preferably, the fat is a triglyceride, most preferably a vegetable triglyceride. Preferably, the emulsifier is selected from polysorbates, monoglycerides, diglycerides, acetic acid esters of monodiglycerides, esters of tartaric acid of monodiglycerides and citric acid esters of monodiglycerides. Preferably, the hydrocoioid is crosslinked or gelled. The entanglement of the hydrocolloids can be carried out using entanglement agents or by a variety of mechanisms. If the hydrocoioid is a protein or polysaccharide containing amino groups, it can be entangled using dialdehydes, such as glutaraldehyde. If the hydrocoioid is a polysaccharide such as sodium alginate, gellan gum or pectin, it can be entangled with multivalent ions, such as calcium or magnesium. The entanglement can be carried out by other mechanisms such as heating, pH adjustment, pressure application or by enzymatic entanglement. Proteins, for example, can be interlaced by subjecting a protein to high pressure, preferably at 5 to 200 bar, and subjecting it to a protein at a temperature that is above the denaturation temperature of the protein. The enzymatic entanglement of proteins can be carried out, for example, with transglutamidase. Based on the hydrocoioid used, one skilled in the art is able to decide which gelation or crosslinking method is used. Preferably, the hydrocoioid is selected from carrageenan. In one aspect, the hydrocoioid is selected from alginate, carrageenan, carboxymethylcellulose (CMC), guar gum, locust bean gum (LBG), xanthan gum, microcrystalline cellulose (MCC), methylcellulose (MC), cellulose esters including hydroxypropylmethylcellulose (HPMC) ), pectin, starch including native and modified starch, pregeiatinized starch and non-pregeiatinized starch, including corn starch, potato, tapioca, wheat and rice, gelatin, agar and combinations thereof. In one aspect, the hydrocolloid is insoluble in brine, particularly at the temperature of use, or a hydrocolloid made insoluble by entanglement or gelation. Preferably, the natural or synthetic polymer is selected from lacquer, polyvinyl acetate, polymethyl methacrylate and its derivatives, any brine-insoluble polymers. In a further preferred aspect, the shell is or comprises or may be formed from the group comprising fats, oils, waxes, resins, emulsifiers or mixtures thereof, which are preferably food grade. Preferably, the hydrophobic shell matrix is selected from the group comprising oils and fats of animal origin, oils of completely hydrogenated vegetable or animal origin, partially hydrogenated vegetable or animal oils, unsaturated, hydrogenated or fully hydrogenated fatty acids, monoglycerides and diglycerides of unsaturated, partially hydrogenated or fully hydrogenated fatty acids, unsaturated, partially hydrogenated or fully hydrogenated esterified fatty acids of monoglycerides or diglycerides, unsaturated, partially hydrogenated or fully hydrogenated free fatty acids, other emulsifiers, animal waxes, waxes of vegetable origin, waxes of mineral origin, synthetic waxes, natural and synthetic resins and mixtures thereof. Oils and fats of animal origin are such as, but not restricted to, beef bait, mutton bait, lamb bait, lard or pork fat, sperm oil. Hydrogenated or partially hydrogenated vegetable oils such as, but not restricted to, canola oil, cottonseed oil, peanut oil, corn oil, olive oil, soybean oil, sunflower oil, safflower oil, oil of coconut, palm oil, linseed oil, stick oil and castor oil. Free fatty acids are such as, but not restricted to, stearic acid, palmitic acid and oleic acid. Other emulsifiers are such as, but not restricted to, polyglycerol esters, sorbitan esters of fatty acids. Originating waxes anima! They are such as, but not restricted to beeswax, lanolin, shell wax or Chinese insect wax. Waxes of vegetable origin are such as but not restricted to carnauba, candelilla, bay berry or sugarcane waxes. Waxes of mineral origin are such as but not restricted to paraffin, microcrystalline oil, ozokerite, ceresin or montana. Synthetic waxes are such as but not restricted to low molecular weight polyolefin, ether-polyol esters and synthetic waxes from the Fisher-Tropsch process. Natural resins are such as rosin, balsam, lacquer and zein. In another preferred aspect, the shell comprises or can be formed of the group comprising hydrocolloids, sodium alginate, gum arabic, gellan gum, starch, modified starch, guar gum, agar gum, pectin, amidified pectin, caragenan, xanthan, gelatin, chitosan, mesquite gum, hyaluronic acid, cellulose derivatives such as cellulose acetate phthalate, hydroxypropyl methylcellulose (HPMC), methyl cellulose, ethyl cellulose and carboxy methyl cellulose (CMC), acrylic methyl copolymers, such as Eudragit®, Psyllium, tamarind, xanthan, locust bean gum, whey protein, soy protein, sodium caseinate, food grade protein, shellac, zein, any synthetic water-soluble or natural polymers, any water-insoluble particles, such as silicon dioxide, titanium dioxide, polymer spheres of synthetic or natural food grade and mixtures thereof. The encapsulated antimicrobial material can be prepared by any suitable method. In a preferred aspect, the encapsulated antimicrobial material is prepared or obtainable by a process selected from spray cooling and fluidized coating. Additional preferred aspects include (a) cooling by sprinkling in fats, waxes or emulsifiers
(b) coating by the fluidized bed with stable lacquer coating of acid, waxes, fats, or emulsifiers, or any other hydrophobic and / or stable coating to acid (c) complex or simple coacervation in interlaced hydrocolloids The encapsulation process is selected preferably from a fluidized bed process, liposome encapsulation, spray drying, spray cooling, extrusion, centrifugal co-extrusion, coacervation and mixtures thereof. Fluidized bed encapsulation and coacervation are the most preferred methods for providing the material of the present invention. In a preferred fluidized bed, the antimicrobial material is coprocessed with an encapsulating material in an aqueous solution or suspension or in a molten state to provide a shell around the antimicrobial material. In the encapsulation of the fluidized bed, the appropriate shell material is typically applied from aqueous solutions or suspensions including HPMC, methylcellulose, microcrystalline cellulose and other cellulose derivatives with or without stearic acid, other fatty acids or other hydrophobic additives. Suitable shell material applied to the molten state include lipids, mono, di or triglycerides, fatty acids, fatty alcohols, waxes or mixtures thereof or any other meltable hydrophobic material. In a preferred coacervation process, an encapsulating material comprising a hydrocolloid or a mixture of hydrocolloids is used to provide the shell. The encapsulation of the antimicrobial material can be carried out by methods that are novel in combination with material 3
antimicrobial and that provide unexpected benefits, for example to the food industry. Encapsulation procedures and encapsulating materials or shell materials are selected depending on the nature of the continuous phase in the food application. The shell material should be water-insoluble if the continuous phase of food application is water-based, and vice versa in order to provide slow and / or delayed release as well as protection / segregation. Suitable encapsulation methods comprise fluidized bed processes, liposome encapsulation methods, spray drying processes, spray cooling processes, extrusion process, coextrusion processes (such as centrifugal coextrusion), coacervation procedures and combinations of the same. In a special double encapsulation process, the present invention provides a microcapsule comprising a solidified hydrophobic shell matrix, a sphere or encapsulated aqueous spheres that are encapsulated in the solidified hydrophobic shell matrix, and antimicrobial material as an active ingredient incorporated in the shell. sphere or encapsulated aqueous spheres. Details of the double encapsulation procedure are described in GB2388581. This dosage form of antimicrobial material is provided by a double encapsulation method for preparing microcapsules, said method comprising the steps of a) providing an aqueous phase and antimicrobial material incorporated in the aqueous phase, b) providing a hydrophobic phase in molten form, c) incorporating or dissolving an encapsulating material or mixtures of encapsulating materials in the aqueous phase or in the hydrophilic phase, d) combining the aqueous phase with the hydrophobic phase and homogenizing or mixing the combined phases to form a water-in-oil emulsion, and ) encapsulating the aqueous phase in the emulsion, thereby converting the liquid aqueous phase into encapsulated aqueous spheres, whereby a dispersion comprising aqueous spheres is formed and the antimicrobial material is incorporated into the aqueous spheres, and f) processing the dispersion obtained in the step e) to form microcapsules wherein the encapsulated aqueous spheres are additionally encapsulated in or by the solidified hydrophobic shell. The encapsulation process of the present invention may also include gelation, entanglement, coacervation, concretion or any other suitable methods. In the above double encapsulation, this results in a dispersion wherein the encapsulated aqueous spheres comprising the active antimicrobial material ingredient are dispersed in the hydrophobic phase. The dispersion is cooled below the melting or dripping point of the hydrophobic phase by any suitable method, which results in the formation of microcapsules. The cooling process can be carried out, for example, by spray cooling or fluidized bed cooling. The microcapsules comprise a number of encapsulated aqueous spheres, which additionally contain the antimicrobial material, and the encapsulated aqueous spheres are further encapsulated in a solidified hydrophobic shell matrix. An advantage of the present invention is that the antimicrobial material is protected by the shell and that the release of the antimicrobial material from the capsules can be controlled. The release rate can be controlled, for example, by the choice and quantity of the shell material. Therefore, the rate of release can be controlled by the fusion of the hydrophobic shell or by the diffusion of water in the capsule and subsequent migration of antimicrobial material to the outside of the capsule. The release rate of antimicrobial material from the capsules can be selected in accordance with the intended use by selecting an appropriate encapsulating material. The release of the antimicrobial material from the capsules of the present invention can be controlled and the release can be initiated in various ways, for example by heat treatment, e.g., by heating, such as in a microwave oven or in an oven , or by freezing, by stress treatment or by any other suitable procedure. The release of the active ingredients from the capsules of the present invention can also be sustained or can occur very slowly. Coacervation is a procedure that works for both water-based and fat-based applications since the shell is entangled and is not soluble in water or fat.
The coacervation process typically involves 1) preparing a suspension of antimicrobial material in an aqueous solution of hydrocolloids, 2) reduce the solubility of hydrocolloids, to cause phase separation and the formation of a third phase rich in hydrocolloid using additives or adjusting the temperature, 3) process the three-phase system in such a way that the newly formed coaservation phase is deposited on the particles of suspended antimicrobial material and finally 4) harden the hydrocolloid shell around the antimicrobial material by adjusting the temperature, adding interlacing chemical or enzymatic or otherwise followed by the isolation of the microcapsules by freeze drying, spray drying, filtration or any other means. The fluidized bed coating is particularly suitable for applications (food) wherein the continuous phase is water, the coating materials include lipid (mono-, di-, tri-glycerides, fatty acids, waxes and mixtures) applied from the molten form , water-insoluble polymers applied from an ethanolic solution (such as zein and lacquer). For applications where the continuous phase is grease, the coating materials include natural, modified or synthetic hydrocolloids (carrageenan, alginate, pectin, locust bean gum (LBG), hydroxypropylmethylcellulose (HPMC), methylcellulose) with or are additives (such as agents) film formers) applied from an aqueous solution or suspension. The particle size of the antimicrobial material should preferably be above 100 μ ??, preferably above 150 μ ??. A double encapsulation in accordance with the present invention is suitable for applications in fat-based foods. The internal phase can be composed of water containing a complex of antimicrobial material / dissolved p-cyclodextrin and any gelled / entangled hydrocolloids or can be composed of glycols (such as ethylene glycol) containing dissolved antimicrobial materials and gelled / interlaced zein. In a liposome encapsulation, the antimicrobial material can be incorporated in the lipid bilayer or in the liposome phase. Spray cooling is a particularly suitable method for water-based applications (food). The antimicrobial material is typically incorporated and suspended in molten lipid (mono, di, triglycerides, fatty acids, waxes and mixtures) and atomized in cold air to form solid particles containing encapsulated antimicrobial material. Spray drying is particularly suitable for grease-based applications (food). The antimicrobial material is typically incorporated and suspended in aqueous solution of hydrocolloids (gum arabic, modified starch, maltodextrins, whey proteins, casein, or the like) with or without additives (such as emulsifiers) and the mixture is atomized in hot air to evaporate the water and form solid particles containing ancapsulating antimicrobial material. Extrusion is a procedure that is typically suitable for grease-based applications (foods) and centrifugal co-extrusion is typically suitable for water-based applications (food). Encapsulation in interlaced hydrocolloid spheres is suitable for both water-based and fat-based food applications. A suspension of an antimicrobial material (alone or in combination with a suitable solvent) is typically prepared in aqueous alginate, pectin, low-ester or any other hydrocolloids "entrelazabas" and thereto is added dropwise or sprayed in a bath of ions of watery calcium. The interlaced spheres or particles containing the encapsulated antimicrobial material are collected by filtration and used as such (wet) or dried by fluidized bed or any other suitable means. Based on the present description, the person skilled in the art is able to select a suitable encapsulation method, as well as the correct type and quantity of shell material to be used in any specific application (food) based on the conditions required to protect and release the antimicrobial material according to the present invention. The encapsulation methods of the present invention are described in some detail below: 1. Fluid bed encapsulation The antimicrobial material is preferably used in the form of a dry powder. If the particle size of the raw antimicrobial material is too fine, the powder can be agglomerated in a suitable equipment using a binder solution (solution of sticky hydrocolloids such as alginate or maltodextrin) in order to obtain a dense powder of particle size between 100-500 μ. The appropriate powder is then introduced into the coating chamber of a fluidized bed microencapsulation unit and is fluidized at an inlet air flow rate of 20-135 cm / s in the lower plate and temperature between 5 and 75 ° C. be used to fluidize the particles. A coating material is then sprayed onto the fluidized bed of antimicrobial using a double fluid nozzle and high pressure atomizing air. In one example, a molten mixture of triglyceride and additives is sprayed onto the antimicrobial powder to form a continuous layer of grease around the individual particle layer as the molten fat spreads and solidifies on the particles. The amount of applied grease can be up to 60%, but usually not less than 15% w / w. In another example, a dispersion or solution of coating material in water and / or ethanol is sprayed onto the fluidized particles and the fluidization air is used to evaporate the solvent or water, which leaves behind a continuous film of coating material on the antimicrobial particles.
Examples of coating material in this case include any hydrocolloids (polysaccharides, proteins, shellac, zein or any other soluble or dispersible coating materials).
2. Liposome Encapsulation Typically, liposomes are prepared using a dehydration-hydration method involving organic solvent, such as that described below. However, solvent-free methods, most suitable for food ingredients, are also available using microfluidization or homogenization devices or by repeated freeze / thaw cycles. A typical procedure for the preparation of antimicrobial material encapsulated in liposome involves the preparation of a solution of 1 g of a bilayer-forming lipid and 100 mg of cholesterol or alpha-tocopherol in a suitable organic solvent and evaporating the solvent to form a film of thin dry lipid on the bottom of the container. After uniform drying of the lipid film 1 liter of water containing antimicrobial material at or above the saturation concentration (the solubility of the antimicrobial material can be increased if desired by the formation of alkali salts) is added to the container and the mixture It is uniformly mixed or homogenized. The resulting multilamellar vesicle suspension (MLV) can be further processed by microfluidization and sonication to form a more homogenous small unilamellar vesicle (SUV). The suspension of antimicrobial material encapsulated in liposome can be added directly to the application or dried by lyophilization or any other appropriate drying procedures.
3. Spray drying The antimicrobial material can be encapsulated in a hydrocolloid matrix by means of spray drying. In a typical procedure, a suspension of aqueous antimicrobial material in which a hydrocolloid or a mixture of hydrocolloids is dissolved (water-soluble polysaccharides, proteins, modified polysaccharides with or without film-forming agents such as oligosaccharides, plasticizers, emulsifiers or other additives) ) is prepared at an almost neutral pH (to minimize the degradation of antimicrobial material). The mixture is then pumped through an atomizer (rotary atomizer, pressure nozzle, double fluid nozzle or any other atomization device) in a co-current or counter-current drying chamber with heated air. The temperature of the heated air is typically between 160 and 200 ° C, it can be as high as 300 ° C, but is preferably in the range of 100-160 ° C. Water evaporation gives a free flowing powder of microcapsules containing dispersed antimicrobial material in the dry hydrocolloid (s) matrix.
4. Spray cooling In spray cooling / cooling / freezing of antimicrobial material, the powder antimicrobial material is dispersed in a melted lipid or lipid mixture (mono-, di-, tri-glycerides, esterified glycerides, animal and vegetable waxes) or mineral, any other meltable material at a temperature between 45 and 125 ° C) with or without the aid of active surface additives. The dispersion is then pumped through an atomizer (rotating atomizer, pressure nozzle, double fluid nozzle, rotating disk or any other atomization device) into a cooling chamber in co-or counter-current with cooled air. The temperature of the cooled air is typically between -10 and 30 ° C, but can be as low as -40oC. The solidification of the lipid gives a free flowing powder of microcapsules containing antimicrobial material dispersed in the crystallized lipid matrix.
5. Extrusion The encapsulation of antimicrobial material by extrusion can be achieved by processing the powder antimicrobial material (preferably of small particle size) together with a molten or plasticized polymer shell material in a twin screw or single screw extruder under pressure, followed by cooling or drying the dough leaving the extruder die and grinding or curling to the appropriate particle size. The polymer mass is melted in the extruder at relatively high temperatures in the presence of a small amount of water, which makes the mass fluidizable. The mass, in which the antimicrobial material is incorporated, is extruded and the cooling results in the transformation of the mass into a vitreous state that is highly impermeable to oxygen and other hydrophobic external agents. Suitable shell materials for extrusion of antimicrobial material include oligosaccharides, polysaccharides, modified polysaccharide, proteins or mixtures thereof with or without the use of plasticizers, emulsifiers or stabilizing additives.
6. - Centrifugal coextrusion The encapsulation of antimicrobial material by centrifugal co-extrusion is a variation of the spray-cooling process. In centrifugal co-extrusion of antimicrobial material, the powder antimicrobial is first dispersed in a molten lipid or mixture of lipids (mono-, di-, tri-glycerides, esterified glycerides, waxes of animal, vegetable or mineral origin, any other meltable material temperature between 45 and 125 ° C) with or without the aid of active surface additives. The dispersion is then pumped through the inner part of a double fluid nozzle while another stream of molten lipid or lipid mixture (the same as before) is pumped through the outer part of the double fluid nozzle. The nozzle is rotated about its axis to break the stream of molten fat into discrete globules, which are solidified by cooled air. The resulting microcapsules are composed of an outer layer of solidified fat that encapsulates a core of solidified fat containing dispersed antimicrobial material.
7. Coacervation The dosage form of antimicrobial material of the present invention can be formed by coacervation. The coacervation of shell material, such as hydrocolloid, is carried out using any suitable coacervation process. The coacervation can be carried out for example by adding salt (s), sugar (s), or other additives, which cause phase separation of the hydrocolloid (s). The coacervation can also be carried out by subjecting the emulsion to heating, cooling, pH change, adding acid or base, which causes phase separation of the hydrocolloid (s). The deposition of the coacervate phase around the aqueous phase is spontaneous and driven by surface tension forces. The coacervate layer can then be subjected to entanglement or hardening by any suitable means, which are known to experts in coacervation. Coating materials suitable for coacervation are selected from the group comprising any mixture of one or many ionic hydrocolloids and one or many amphoteric hydrocolloids, such as any mixture of polysaccharides and proteins, gelatin / gum arabic, gelatin / CMC, any proteins / hydrocolloids ionics, any combination of hydrocolloids and solubility reducing agent such as salts, sugars, acids or bases.
8. Double encapsulation In accordance with a special aspect of the present invention, the suspension of antimicrobial material is double encapsulated in microcapsules. In that case, the antimicrobial material is first incorporated (suspended) in an aqueous phase containing encapsulating material, such as hydrocolloid or any other suitable encapsulating material or mixture thereof, and the aqueous phase is encapsulated, for example by gelation, interlacing, coacervation, concretion or by any other suitable means, and the resultant encapsulated aqueous sphere or spheres is / are further encapsulated in a solidified hydrophobic shell material. In a preferred aspect, the encapsulated antimicrobial material is encapsulated by one and only one shell. In this regard, the present invention provides an antimicrobial material in an encapsulated form, comprising (i) a shell comprising an antimicrobial material and (ii) a shell of encapsulating material, wherein the shell of matter! encapsulant in impermeable to the antimicrobial material and in which the antimicrobial material is encapsulated only by the shell (i). A hydrophobic shell material is selected based on the desired properties of the capsules, eg based on the intended use of the capsules, storage temperature, etc. The hydrophobic shell material should have a melting point above 45 ° C so it can be stored at room temperature, in general any hydrophobic material can be used if the capsules are stored below the melting temperature of the hydrophobic material . In this request, molten form means that the hydrophobic phase is at the lowest temperature at which the hydrophobic phase is sufficiently fluid to drip, as determined by the test method AST D 566 or D 265. In a preferred aspect, the shell of the material Encapsulated antimicrobial is able to resist injection into or on the food. In a preferred aspect, the shell of the encapsulated antimicrobial material is capable of withstanding a pressure of more than 1.5 bar, for example of more than 2.0 bar, for example of more than 3.0 bar. In a preferred aspect, the shell of the encapsulated antimicrobial material is able to withstand a shear force greater than that typically encountered during injection. As described herein, the shell of the encapsulated antimicrobial material can be selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material or to provide release under predetermined conditions. In addition, the shell of the encapsulated antimicrobial material can be selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material. Preferably, the degeneracy to be prevented is by one or more factors selected from thermal degradation, pH-induced degradation, degradation by protease and formation of glutaion adduct. The inventors of the present invention have found that the provision of an encapsulated antimicrobial material in which the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material or to provide release under predetermined conditions is advantageous regardless of the manner in which the antimicrobial material is encapsulated. which the encapsulated antimicrobial material is contacted with a food. For example, the encapsulated antimicrobial material may be contacted with a food (or other material) by means other than injection or stirring. In other words, the inventors herein have provided an encapsulated antimicrobial material in which the antimicrobial material is released in a sustained manner or when the encapsulated antimicrobial material is placed under predetermined conditions. Therefore, in a further aspect (the aspect of "encapsulated material"), the present invention provides an antimicrobial material in an encapsulated form, comprising a core of antimicrobial material and shell of encapsulating material, wherein the shell of encapsulating material It is impermeable to the antimicrobial material. This preservation system has the benefits of maximizing the potential of the antimicrobial material (such as nisin) that can be added to the food, offering no impact of taste, economy, ease of use, ease of manufacture and stability. In some aspects of the invention, it can also be described as "natural" for food labeling purposes. In this and other aspects of the invention, the term "encapsulation" is understood to be the packaging of solid particles or liquid droplets of active ingredient (or particles or droplets containing the active ingredient) within a secondary material to completely surround the solid particles or drops of liquid with a protective or functional shell material. This contrasts with the loose use of the term encapsulated to refer to simple coating. For example, Cahill et al. They teach the nisin coating are a porous matrix of aiginate. The external material can diffuse freely in the aiginate matrix and the coated nisin can easily diffuse through the large pores of the matrix. This is not "encapsulation" in the current sense. Each of the preferred aspects described herein are applicable to the appearance of the encapsulated material of the invention. Particularly preferred aspects include • the antimicrobial material is an antibacterial material. • the antimicrobial material is a bacteriocin. • the antimicrobial material is at least nisin. • The shell is selected to provide sustained release of the animicrobial material from the encapsulated antimicrobial material. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material.
• The shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation. • the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a marinade. • the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes. • the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products.
• The encapsulated material is used in protection of microbial putrefaction of a raw poultry product. Particularly preferred aspects further include • the antimicrobial material is an antifungal material • the antimicrobial material is at least natamycin • the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material. • The shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation. • the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a confectionery product. Highly preferred aspects of all aspects of the invention and in particular of the encapsulated material aspect include • the antimicrobial material is at least nisin. • The shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation. • Antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes. • the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products.
• The encapsulated material is used in microbial rot protection of a raw poultry product. • the antimicrobial material is at least nisin; and the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, degradation with protease and formation of glutathione adduct. • the antimicrobial material is at least nisin; and the antimicrobial material provides a bactericidal or bactenostatic effect with respect to Listeria monocytogenes. • the antimicrobial material is at least nisin; and the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, degradation with protease and formation of glutathione adduct; and the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more selected factors of thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation; and the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. • the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes; and the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. • the antimicrobial material is at least nisin; and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation; and the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes. • the antimicrobial material is at least nisin; and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation; and the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. • the antimicrobial material is at least nisin; and the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes; and the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, degradation with protease and formation of glutathione adduct; and the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes; and the encapsulated material is used in the protection of microbial putrefaction of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. • the antimicrobial material is at least nisin; and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation; and the antimicrobial material provides a bactericidal or bacteriostatic effect with respect to Listeria monocytogenes; and the encapsulated material is used in the protection of microbial spoilage of a selected food of raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. Additional highly preferred aspects of all aspects of the invention and in particular the appearance of encapsulated material include • the antimicrobial material is at least natamycin. • The shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material. • the shell is selected to prevent, reduce or inhibit the degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation • the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a confectionery product. • the antimicrobial material is at least natamycin, and the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. • the antimicrobial material is at least natamycin, and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material.• the antimicrobial material is at least natamycin, and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, degradation with protease and formation of adduct from glutathione • the antimicrobial material is at least natamycin; and the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a confectionery product. • the antimicrobial material is at least natamycin; the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material. • the antimicrobial material is at least natamycin; the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, degradation with protease and glutathione adduct formation. • the antimicrobial material is at least natamycin, and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material, and the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a confectionery product. • the antimicrobial material is at least natamycin, the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, degradation with protease and formation of glutathione adduct , and the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a confectionery product. • the antimicrobial material is at least natamycin and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by pH-induced degradation. • the antimicrobial material is at least natamycin and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by pH-induced degradation, and the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. • the antimicrobial material is at least natamycin and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by pH-induced degradation, and the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably food is a confectionery product. • the antimicrobial material is at least natamycin and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by pH-induced degradation, and the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material, and the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a food, preferably the food is a confectionery product. • the antimicrobial material is at least natamycin and the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by pH-induced degradation, and the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material, and the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material in contact with a confectionery product.
Food In one aspect, the invention is to improve the use of antimicrobial materials in the food industry and, as a consequence, the shell of the present invention must be made of a physiologically acceptable material suitable for addition to a food product. The shell provides protection for the antimicrobial material and must be effective in substantially retaining the antimicrobial material within the shell during the processing of food products. Once introduced into a food product, the shell should be effective to provide slow or delayed release of the antimicrobial material encapsulated in the food product. Most preferably, the antimicrobial material of the present invention has a shell that is effective to protect the encapsulated antimicrobial material against degradation by conditions prevailing in the production of a product to which encapsulated antimicrobial material is added, and / or to protect food ingredients against unwanted attack by antimicrobial material at the wrong time, as well as to provide release of antimicrobial material in the finished product. The term "food", as used herein, in the present application and in the claims, generally refers to edible products and beverages from the food and feed industry. The edible products in question are mainly nutritious and / or enjoyable products that require preservation for storage between the time of production and the final use. Many foods can be protected by the present invention. Typical foods are raw meat, cooked meat, raw poultry products, cooked poultry products, raw seafood products, cooked seafood products, [raw or cooked meat, poultry and seafood products, ready-to-eat meals, sauces for pasta, pasteurized soups, mayonnaise, salad dressings, marinades, oil-in-water emulsions, margarines, low-fat accounting products, water-in-oil emulsions, dairy products, spreadable cheeses, processed cheese, dairy desserts, flavor milks , cream, fermented milk products, cheese, butter, condensed milk products, ice cream mixes, soy products, pasteurized liquid egg, confectionery products (such as soft buns), confectionery, fruit products and stuffed foods based on fat or contain water. Additional preferred foods, particularly when the antimicrobial material is natamycin are salad dressings, acidic dairy products (including natural cheese, cottage cheese, acidified cheese, cream cheese, yogurt, sour cream, processed cheese), fruit juices, acidic beverages, beverages alcoholic beverages (including wine and beer), chilled dough and cooked or uncooked baked goods, fillings and dairy covers for baked goods, covers and surface coatings for baked goods and other heat processed items, condiments, spreadable sauces for snacks , purées, canned gherkins, marinated, marinated meat or poultry, breaded meat or poultry, covers and bases on pizzas, fast food products, equipment to make snacks or meals, equipment to make confectionery products, pet food, food for broilers and any other acidic food products, heat processed and / or fermented with mushrooms. A particularly preferred preserved food product is a sliced or sliced pastry product, especially sliced bread, wherein the encapsulated material such as natamycin has been incorporated into the dough prior to cooking and provides preservation of the confectionery product after baking. A special benefit is provided by a preferred embodiment of the invention when the encapsulated antimicrobial material is included in a dough before the baking of a high pastry product with yeast since the yeast is protected by the encapsulating shell against direct contact with the antimicrobial material during the lift. further, the encapsulated antimicrobial material is preferably protected from the heat of baking by the shell. The antimicrobial material is degraded by exposure to heat. During baking, which is typically formed at temperatures ranging from 180 to 300 ° C, the degradation of the antimicrobial material would significantly reduce the level of active antimicrobial material in the finished confectionery product. By selecting an encapsulating material that has sufficient heat stability, the thermal degradation of the antimicrobial material can be substantially reduced. During baking and / or after baking, the shell releases the antimicrobial material so that the finished product is effectively protected against fungal attack. A preferred use of the present invention comprises the use of the form of the bulk antimicrobial material for bread, which is to be sliced for sale. The antimicrobial material released from inside the shell of the capsule in the finished product protects the individual bread slices against fungal attack. Sliced bread is a very convenient food product for consumers. However, slicing provides an additional procedural step in the production, and one that is typically performed after the bread has cooled after baking when the product is very susceptible to fungal attack. When slicing is performed, contamination can take place and as a result, the sliced product will show fungal growth between the slices during storage. Slicing of bread exposes a much larger surface area of the bread to contamination particularly by molds. The copending patent application US 10 / 765,210, filed on January 28, 2004 and included herein by reference, describes the protection of baked goods by spraying the surface of the products with antimicrobial material and therefore increases shelf life. of the product. However, it is impossible to apply antimicrobial material between slices of sliced bread. The present invention provides a solution to the problem of protection of bread sliced by antimicrobial material. Another preferred preserved food product comprises an acidic food product, in which the material of the present invention such as natamycin has been incorporated. In a preferred aspect, the food is a confectionery product. Many antimicrobial materials such as natamycin in solution are very stable at neutral pH but are easily degraded, especially at room temperature when the pH increases above pH 10 or below pH 4.5, and especially below pH 4. Thus, for example, antimicrobial materials included in acidic products will gradually degrade and consequently leave the product unprotected during storage and use. The rate of degradation of antimicrobial material increases as the temperature increases. Many acidic products, such as salad dressings and condiments are stored at room temperature and used for a long period of time even after opening the package. Acid drinks such as fruit juices can be stored at room temperature and can be opened to fungal attack. Marinades and marinated meat and poultry are typically stored for a long time at room temperature. Many acidic dairy products are stored at room temperatures or cooled and can be rotten by fungal growth. When the encapsulated antimicrobial material is added to such products, the encapsulation protects the enclosed antimicrobial material and allows it to slowly diffuse into the product to replace any degraded antimicrobial material thereby keeping the amount of antimicrobial material active at an adequate antifungal level in the product. . The encapsulated antimicrobial material of the present invention provides similar benefits in other acidic products, especially those that are stored at room temperature. Foodstuffs, which are especially suitable for preservation by the dosage form of the novel antimicrobial material of the present invention include fatty acid-containing products such as salad dressings and acidic dairy products (natural cheese, cottage cheese, acidified cheese, cheese cream, yogurt, sour cream). Many of these products can be preserved with antimicrobial material in an unencapsulated form and will generally remain well if stored in refrigeration. However, if stored at room temperature, degradation of the antimicrobial material is a problem. This problem is solved with the encapsulated antimicrobial material of the present invention. In the United States, salad dressings are generally cold processed, in which case yeast and mold contaminants are potential spoilage contaminants. Storage contamination of ambient temperature and low pH causes degradation of fast antimicrobial material. If the encapsulated antimicrobial material, which is added when the dressings are first made, does not kill polluting yeasts quickly, and if any mold spores are present (these are not normally killed by antimicrobial material), there is potential for fungal growth / rot. once the levels of antimicrobial material fall. By using the encapsulated antimicrobial material of the present invention, acidic food products can be stored at room temperature for up to 12 months. Preferred methods for encapsulation for acidic food products comprise coacervation and fluidized bed encapsulation. The encapsulating shell can also be designed to protect the antimicrobial material against any heat during the processing of the acidic food product, such as pasteurization at temperatures of typically 60 to 120 ° C and more frequently 60 to 95 ° C. Another type of food product that derives great benefits from the present invention is fruit juice and acidic beverages. Benefits are also derived for processed fruits, low pH sauces such as tomato sauces and purees, hot sauces, spreadable sauces for snacks, canned pickles, etc., alcoholic beverages such as wine or beer and the like. These liquid products may contain fat (milk drinks with acidified fruits, etc.). They can be pasteurized. The combination of pasteurization at low pH, but more importantly acid pH and storage at room temperature results in the degradation of unprotected antimicrobial material. If post-processing contamination with yeast or molds has occurred, heat-resistant mold spores (eg, Byssochiamys Talaryomyces) or yeast ascospores survive processing, fungal growth will occur once levels of antimicrobial material be degraded. Animal feed products such as dog and cat food or broiler feed are often hot processed during the production thereof and then stored at room temperature. The encapsulated heat stable antimicrobial material of the present invention can be conveniently used to protect said food products. In liquid products such as juices or wines, the shell material should preferably be made from a material that does not distribute the clarity of the liquid. When the antimicrobial material is added in the form of novel capsules of the present invention, the shell will slowly dose small amounts of antimicrobial material and keep the liquid products free of microbial growth, such as fungi, for prolonged periods (3-9 months) of storage. at room temperature. The encapsulation provides a special benefit for heat-treated acid liquids since the shell protects the antimicrobial material against heat and acid attack. The antimicrobial material is a preservative that can also be used to take advantage of confectionery products. Most baked goods are susceptible to mold rot due to aerial contamination with mold spores after baking. Propionate is commonly incorporated in bread and other yeast lifting masses as an anti-mildew agent. The anti-yeast activity of propionate is much weaker compared to the anti-mold activity in these masses. Although propionate has a slight inhibitory effect against baking yeast, this is acceptable. The antimicrobial material can not be used in this way because it is strongly active against both yeasts and molds. The encapsulation of the antimicrobial material prevents the activity of the antimicrobial material against the baking yeast until after the elevation is complete. It also protects the antimicrobial material during the baking process. This is particularly useful for products such as sliced breads that have a large surface area exposed to air pollution. In a preferred aspect, the food is selected from raw meat, cooked meat, raw poultry products, cooked poultry products, raw seafood products, cooked seafood products [meat raw and cooked poultry and seafood products] and raw foods or cooked, subject to bacterial surface growth. In a preferred aspect the food is raw meat. In a preferred aspect, the food is selected from raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. In a preferred aspect, the food is selected from raw poultry products and cooked poultry products. In a preferred aspect, the food is a product of raw poultry. In a preferred aspect, the food comprises whole meat muscle. Those skilled in the art will appreciate that the term "cooked" means a food product that has undergone some degree of cooking (either partial or complete). Those skilled in the art will appreciate that the cooked products of the present invention may be subjected to further cooking after contacting the encapsulated material of the present invention. In a preferred aspect, the cooked products of the present invention are subjected to further cooking after contact with the encapsulated material of the present invention. The subsequent firing was the release of antimicrobial material from the encapsulated product and consequently the activation of the protective effect of the antimicrobial material.
Additional components The encapsulated antimicrobial material may contain one or more components in addition to the core of an antimicrobial material and the encapsulant matenal shell. These or more additional components may or may not be encapsulated within or by the shell along with the antimicrobial material. In other words, the additional components can be encapsulated within or by the shell along with the antimicrobial material or they can be "outside" the shell. When one or more additional components are provided, a combination of the above is contemplated (one component may be inside the shell and another component outside the shell). Typically, the encapsulated antimicrobial material will not be introduced into or onto the food alone. Therefore, in one aspect the encapsulated antimicrobial material is introduced into or onto the food in a vehicle. Preferably the vehicle is or comprises brine. The density of the encapsulated antimicrobial material must match the density of the vehicle (such as brine) to prevent separation or settling of the encapsulated antimicrobial material, avoiding any distribution of encapsulated antimicrobial material during injection or stirring. Therefore, in a preferred aspect the vehicle and the encapsulated antimicrobial material have substantially the same density. The equalization of the vehicle density and the encapsulated antimicrobial material can be achieved by careful selection of the vehicle and encapsulated antimicrobial material. Alternatively, it can be achieved by modifying the encapsulated antimicrobial material to have substantially the same density as the vehicle, or by modifying the vehicle to have substantially the same density as the encapsulated antimicrobial material. The encapsulated antimicrobial material can be modified by contacting the encapsulated antimicrobial material with oil, such as brominated oil. The vehicle can be modified by inclusion of an additional component such as xanthan gum. The vehicle may contain one or more additional components. However, in some aspects, the vehicle does not contain additional components or contains no additional components that materially affect the properties of the composition. In a preferred aspect, the vehicle may comprise an emulsifier. Preferably, the emulsifier is selected from polyoxyethylene sorbitan esters (E432-E436) otherwise known as polysorbates (e.g., Tween 80, Tween 20), monoglycerides, diglycerides, acetic acid esters of monodiglycerides, esters of tartaric acid of mono- and diglycerides and citric acid esters of monodiglycerides. The encapsulated antimicrobial material may contain one or more additional components. However, in some aspects the encapsulated antimicrobial material contains no additional components or contains no additional components that materially affect the properties of the composition. In a preferred aspect, the encapsulated antimicrobial material may further comprise an extract obtained or obtainable from a plant of the Labiatae family. Optionally in this aspect and particularly when the antimicrobial material consists of nisin, the composition comprises carvacrol in an amount of less than 0.05% by weight based on the composition and carvone in an amount of less than 15% by weight based on the composition . Compositions comprising an antimicrobial material and an extract obtained from or obtainable from a plant of the Labiatae family are described in British patent application No. 0323335.0. Each of the teachings of GB 0323335.0 is applicable to the present system. In this aspect, preferably the extract obtained or obtainable from a plant of the Labiatae family is not encapsulated within or by the shell together with the antimicrobial material. In a preferred aspect, the extract contains carvacrol in an amount of less than 0.075% by weight based on the composition, preferably in an amount of less than 0.04% by weight based on the composition, most preferably in an amount of less than 0.02% by weight based on the composition. In a preferred aspect, the extract contains carvone in an amount of less than 0.075% by weight based on the composition, preferably in an amount of less than 0.04% by weight based on the composition, most preferably in an amount of less than 0.02% by weight based on the composition. In a preferred aspect, the extract contains thymol in an amount of less than 0.1% by weight based on the composition, preferably in an amount of less than 0.075% by weight based on the composition, most preferably in an amount of less than 0.0% by weight based on the composition. In one aspect, the extract used is obtained from a plant of the Labiatae family. One skilled in the art will appreciate that by the term "extract" or "extracts" is meant any constituent of the plant that can be isolated from the whole plant. In one aspect, the extract used in the present invention is obtainable from a plant of the Labiatae family. One skilled in the art will appreciate that a obtainable extract of a plant can be obtained from a plant or can be isolated from a plant, identified and then obtained from an alternative source, for example, by chemical synthesis or enzymatic production. For example, the extract can be produced by eukaryotic or prokaryotic fermentation, by a method of genetic manipulation. The applicant of the present has recognized that the products present in a plant of the Labiatae family can synergistically increase the activity of antimicrobial material, preferably a bacteriocin. These products can be obtained from any source and will fall within the scope of the present invention. The invention comprises the use of a combination of an antimicrobial material, for example a bacteriocin and in particular nisin, and of the Labiatae family of plants, such as rosemary. { Rosmarinus officinalis) or sage (Salvia officinalis) that together give an increased control of gram-positive bacteria in a food system. The extracts responsible for synergy in the present invention preferably refer to extracts from the family of Labiatae plants that have been selectively extracted ("deodorized extracts") to increase their content of phenolic diperpene (such as carnosol and carnosic acid), tripterpene content phenolic (such as ursolic acid, betulinic acid and oleanolic acid) or rosmarinic acid content. These deodorized extracts can be distinguished by their high phenolic diterpene content (for example greater than 3.5% by weight) and their low level (less than 1% by weight) of flavor-inducing compounds of essential oils and oleoresins of the plant that are used as flavors or fragrances. Essential oils are typically extracted by simple steam distillation of the plant material. In a preferred aspect, the extract is a deodorized extract. Preferably, the extract (deodorized) contains from 1.0 to 70% by weight of phenolic diperpenes, preferably 3.5 to 70% by weight of phenolic diterpenes and less than 1% by weight of essential oil. In a preferred aspect, the extract is selected from phenolic diterpenes, phenolic triterpenes and rosmarinic acid. In a preferred aspect, the extract is or comprises a phenolic diterpene. Preferably, the phenolic diterpene is selected from carnosic acid, carnosol, methylcarnosic acid and mixtures thereof. Preferably the phenolic diterpene is selected from carnosic acid and
carnosol. In a preferred aspect, the extract contains phenolic diterpenes in an amount of more than 1.0% by weight, based on the composition, preferably in an amount of more than 2.0% by weight, based on the composition, most preferably in an amount of more than 3.0% by weight, based on the composition, most preferably in an amount of more than 3.5% by weight, based on the composition. In a highly preferred aspect, the extract contains one or more phenolic triterpenes. Preferably the phenolic triterpenes are selected from betulilic acid, oleanolic acid and ursolic acid. In a preferred aspect, it is or comprises a phenolic triterpene. Preferably, the phenolic triterpene is selected from betulinic acid, oleanolic acid and ursolic acid. In a preferred aspect, the extract is or comprises rosmarinic acid. In a preferred aspect, the plant of the Labiatae family is selected from rosemary, sage, oregano, marjoram, mint, balsam, savory and thyme. In a preferred aspect, the plant of the Labiatae family is selected from rosemary, sage, oregano, marjoram, mint, balsam and savory. It will be understood that these names cover all the species and varieties of plants known by these names. In a preferred aspect, the plant of the family Labiatae is selected from rosemary (Rosmarinus officinalis L.), sage (Salvia offícinalis L), oregano (Origanum vulgare L), marjoram [Origanum marjorana L), mint (Mentha spp.), balsam (Melissa officinalis L), savory (Satureia hortensis), and thyme (Thymus vulgaris L). In a preferred aspect, the Labiatae family plant is selected from rosemary (Rosmarinus officinalis L), sage (Salvia officinalis L), oregano (Origanum vulgare L), marjoram (Origanum marjorana L), mint (Mentha spp.), Balsam (Melissa officinalis L), and savory (Satureia hortensis),
In a preferred aspect, the plant of the Labiatae family is rosemary. In a preferred aspect, the extract contains flavor-inducing compounds and / or essential oils in an amount of less than 1% by weight based on the extract. In a preferred aspect, the extract contains flavor-inducing compounds and / or essential oils in an amount of less than 1% by weight based on the composition. Typically, the flavor-inducing compounds and / or essential oils are camphor, verbenone, borneol and alpha-terpineol. In a preferred aspect, the combined amount of camphor present in the extract is less than 1% by weight (preferably less than 0.2% by weight, most preferably less than 0.15% by weight, most preferably less than 0.1% by weight) based on in the extract. In a preferred aspect, the combined amount of verbenone present in the extract is less than 1% by weight (preferably less than 0.2% by weight, most preferably less than 0.15% by weight, most preferably less than 0.1% by weight) based on in the extract. In a preferred aspect, the combined amount of borneol present in the extract is less than 1% by weight (preferably less than 0.2% by weight, most preferably less than 0.15% by weight, most preferably less than 0.1% by weight) based on in the extract. In a preferred aspect, the combined amount of alpha-terpineol present in the extract is less than 1% by weight (preferably less than 0.2% by weight, most preferably less than 0.15% by weight, most preferably less than 0.1% by weight) based on the extract. In a preferred aspect, the combined amount of camphor, verbenone, borneol and alpha-terpineol present in the extract is less than 1% by weight (preferably less than 0.2% by weight, most preferably less than 0.15% by weight, most preferably less than 0.1% by weight) based on the extract. In a preferred aspect, the encapsulated antimicrobial material further comprises a chelator. Preferably the chelator is selected from EDTA, citric acid, monophosphates, diphosphates, triphosphates and polyphosphates. An additional suitable chelator is taught in US 5573801 and includes carboxylic acids, polycarboxylic acids, amino acids and phosphates. In particular, the following compounds and their salts may be useful. Acetic acid, adenine, adipic acid, ADP, alanine, B-alanine, albumin, arginine, ascorbic acid, asparagine, aspartic acid, ATP, benzoic acid, n-butyric acid, casein, citraconic acid, citric acid, cysteine, dehydracetic acid, Desferri-ferricrisine, Desferri-ferricrome, Deferrifrioxamine E, 3,4-dihydroxybenzoic acid, diethylenetriaminepentaacetic acid (DTPA), dimethylglyoxime, 0,0-dimethylpurpurogaline, EDTA, formic acid, fumaric acid, globulin, gluconic acid, glutamic acid, glutaric acid, glycine, glycolic acid, glycylglycine, glycic sarcosine, guanosine, histamine, histidine, 3-hydroxy flavone, inosine, inosine triphosphate, iron free ferricrome, isovaleric acid, itaconic acid, kojic acid, lactic acid, leucine, lysine, acid maleic, malic acid, methionine, methyl salicylate, nitrilotriacetic acid (NTA), omitin, orthophosphate, oxalic acid, oxistearin, B-phenylalanine, phosphonic acid rich, phytate, pimelic acid, pivalic acid, polyphosphate, proline, propionic acid, purine, pyrophosphate, pyruvic acid, riboflavin, salicylaldehyde, salicylic acid, sarcosine, serine, sorbitol, succinic acid, tartaric acid, tetrametaphosphate, thiosulfate, threonine, trimetaphosphate , triphosphate, tryptophan, uridine diphosphate, uridine triphosphate, n-valeric acid, valine and xanthosine. Many of the sequestering agents are useful in food processing in their salt forms, which are commonly alkali metal or alkaline earth metal salts such as sodium, potassium or calcium salts or quaternary ammonium salts. Sequestering compounds with multiple valencies can be used beneficially to adjust the pH or selectively introduce or abstract metal ions eg in a food system coating. Chelators of additional information are described in T. E.
Furia (Ed.), CRC Handbook of Food Additives, 2nd Ed., Pp. 271-294 (1972, Chemical Rubber Co.), and M. S. Peterson and A.. Johnson (Eds.), Encyclopaedia of Food Science, pp. 694-699 (1978, AVI Publishing Company, Inc.), both articles incorporated herein by reference. The term "chelator" is defined as organic or inorganic compounds capable of forming coordination complexes with metals. Also, as the term "chelator" is used herein, it includes molecular encapsulating compounds such as cyclodextrin. The chelator can be inorganic or organic, but preferably is organic. Preferred chelators are non-toxic to mammals and include aminopolycarboxylic acids and their salts such as ethylenediaminetetraacetic acid (EDTA) or their salts (particularly their di- and tri-sodium salts), and hydroboxylic acidic acids and their salts such as citric acid. However, it is believed that non-citric acid, non-citrate hydrocarboxylic acid chelators are also useful in the present invention such as acetic acid, formic acid, lactic acid, tartaric acid and its salts. As noted above, the term "chelator" is defined herein as a synonym for sequestering agent and is also defined as including encapsulating compounds such as cyclodextrin. Cyclodextrins are cyclic carbohydrate molecules that have six, seven or eight glucose monomers exposed in a donut-shaped ring, which are denoted alpha, beta or gamma cyclodextrin, respectively. As used herein, cyclodextrin refers to both unmodified and modified cyclodextrin monomers and polymers. Molecular cyclodextrin encapsu tators are commercially available from American Maize-Products of Hammond, Ind. Cyclodextrins are further described in Chapter 11 entitled "Industrial Applications of Cyclodextrin" by J. Szejtli, page 331-390 of Inclusion Compounds, Vol. III (Academic Press, 1984) chapter that is incorporated herein by reference. When the antimicrobial material is natamycin, the cyclodextrins are particularly preferred. Cyclodextrins improve the solubility of natamycin. Preferably, the chelator increases the antimicrobial activity and / or antimicrobial spectrum of the bacteriocin. Most preferably the chelator increases the antimicrobial activity and / or antimicrobial spectrum of the bacteriocin with respect to gram-negative bacteria and other microorganisms. The inventors of the present have found that providing a chelator is particularly effective in view of the increased antimicrobial activity and / or antimicrobial spectrum of the bacteriocin provided. The increase is possible regardless of the manner in which the encapsulated antimicrobial material is supplied or the nature of the shell of the encapsulated antimicrobial material. Therefore, in a further aspect, the present invention provides an antimicrobial material in an encapsulated form comprising (a) a core of (i) an antimicrobial material and (ii) a chelator; and (b) a shell of encapsulating material.
In a preferred aspect, the encapsulated antimicrobial material further comprises an organic acid, a salt thereof or a mixture thereof. Particularly preferred organic acids are lactic acid and acetic acid. Preferably, the organic acids are provided in their salt form such as the sodium sai or potassium salt of the respective acid. Highly preferred organic acid salts are sodium lactate (L-sodium lactate), potassium lactate (L-potassium lactate), sodium di-acetate and mixtures thereof. Particularly preferred mixtures are mixtures of sodium lactate (L-sodium lactate) and sodium di-acetate; and mixtures of sodium lactate and potassium acetate. Suitable salts of organic acids (and mixtures thereof) are available from Purac, The Netherlands, under the name PURASAL®. Each of the preferred aspects described herein are applicable to this aspect of the invention. Particularly preferred aspects include • where the shell of encapsulating material is impermeable to the antimicrobial material. • The shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. • the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material. • the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the bacteriocin • the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the bacteriocin with respect to gram-negative bacteria and other microorganisms. • the chelator is selected from EDTA, citric acid, monophosphates, diphosphates, triphosphates and polyphosphates. • the antimicrobial material is an antibacterial material. • the antimicrobial material is a bacteriocin. • the antimicrobial material is at least nisin. • the antimicrobial material is an antifungal material. • the antimicrobial material is at least natamycin.
Procedure The encapsulated antimicrobial material can be introduced into or onto the food by any suitable method. For example, it can be introduced into or on the food by sprinkling, dipping, injecting, stirring or mixing (in the food matrix). The encapsulated antimicrobial material can be introduced into or onto the food by (a) injecting the antimicrobial material encapsulated in the feed or (b) stirring the antimicrobial material encapsulated with the feed. In one aspect, the encapsulated antimicrobial material is introduced into the food by injecting the antimicrobial material encapsulated in the food. In one aspect, the encapsulated antimicrobial material is introduced into or onto the food by stirring the antimicrobial material encapsulated with the food. As indicated herein, the encapsulated antimicrobial material may be introduced into or onto the food by means other than injection or stirring. For example, the encapsulated antimicrobial material can be incorporated into a marinade. Marinated meat can be prepared in two ways: 1) a surface treatment (such as, but not limited to, adding marinade to raw meat followed by packing under gas or vacuum) or 2) forced incorporation onto the marinated / brine by physical means (such as, for example but not limited to, stirring or injection). The teachings on the practice of food injection or stirring of food can be found in WO 00/62632.
Additional Aspects Additional aspects of the present invention will be described in the following numbered paragraphs: 1. A dosage form of antimicrobial material (preferably natamycin) comprising microcapsules wherein the antimicrobial material (preferably natamycin) is encapsulated within a physiologically acceptable shell for provide a product of protected food preservative antimicrobial (preferably natamycin) material. 2. A dosage form of antimicrobial material (preferably natamycin) in accordance with paragraph 1, wherein said shell is effective to retain substance said antimicrobial material (preferably natamycin) within the shell during processing of the food product. 3. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 1, wherein said shell is effective to provide slow or delayed release of the encapsulated antimicrobial material (preferably natamycin) in the food product. 4. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 1, wherein said shell is effective to protect the encapsulated antimicrobial material (preferably natamycin) against degradation by prevailing conditions in the production of a product to which the encapsulated antimicrobial material (preferably natamycin) is added and to provide release of antimicrobial material (preferably natamycin) in the finished product. 5. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 1, wherein said encapsulation is provided by a method selected from a fluidized bed process, liposome encapsulation, spray drying, spray cooling, extrusion. , co-extrusion, coacervation and combinations thereof. 6. A dosage form of antimicrobial material (preferably natamycin) in accordance with paragraph 1, wherein the shell is made of a material selected from the group consisting of hydrophobic materials, hydrocolloid materials and mixtures or combinations thereof. 7. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 6, wherein said hydrophobic matenal is chosen from lipids and resins including fatty acids, fats, oils, emulsifiers, fatty alcohols, waxes and mixtures or combinations thereof. 8. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 7, wherein said hydrophobic material is selected from the group consisting of food grade animal oils and fats, fully hydrogenated vegetable or animal oils. , partially hydrogenated vegetable or animal oils, unsaturated, hydrogenated or fully hydrogenated fatty acids, monoglycerides and diglycerides of unsaturated, hydrogenated or fully hydrogenated fatty acid, unsaturated fatty acids, partially hydrogenated, fully hydrogenated of monoglycerides or diglycerides, unsaturated free fatty acids, partially hydrogenated, completely hydrogenated, other emulsifiers, waxes of animal origin, waxes of vegetable origin, waxes of mineral origin, synthetic waxes, natural and synthetic resins and mixtures thereof. 9. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 6, wherein said hydrocolloid comprises a soluble or dispersible coating material selected from food-grade gums, polysaccharides, proteins, lacquer and mixtures or combinations thereof. same. 10. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 9, wherein said hydrocolloid is selected from cellulose derivatives including hydroxypropylmethylcellulose, cellulose acetate phthalate, carboxymethylcellulose, methylcellulose and microcrystalline cellulose, sodium alginate, gum arabic, gelano gum, guar gum, agar gum, pectin, amidified pectin, carrageenan, gelatin, chitosan, mesquite gum, hyaluronic acid, methylacrylic copolymers, such as Eudragit®, Psyllium, tamarind, xanthan, locust bean gum, Welano gum, zein, lacquer, whey protein, soy protein, sodium caseinate, natural or synthetic water soluble polysaccharides, proteins and other hydrocolloids, with or without fatty acids, fatty alcohol, plasticizers including glycerol, polyethylene glycol and other hydrophilic alcohols of low molecular weight, or combinations of any of said hydrocolloids. 11. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 1, wherein the shell is provided by co-processing antimicrobial material (preferably natamycin) with an encapsulating material, which is in aqueous solution or suspension or lipid in a molten state. 12. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 11, wherein the antimicrobial material (preferably natamycin) is an aqueous suspension or comprises a dry powder. 13. A dosage form of antimicrobial material (preferably natamycin) in accordance with paragraph 1, comprising microcapsules having a solidified hydrophilic shell matrix, encapsulated aqueous spheres that are additionally encapsulated in the solidified hydrophilic shell matrix, and antimicrobial material (preferably natamycin) incorporated in the encapsulated aqueous spheres. 14. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 1, wherein the percentage of active antimicrobial material material (preferably natamycin) in the product of protected antimicrobial material (preferably natamycin) is from 1 to 80% in weigh. 15. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 14, wherein the percentage is between 15 and 50% by weight. 16. A dosage form of antimicrobial material (preferably natamycin) according to paragraph 15, wherein the percentage is between 30 and 40% by weight. 17. A method for preparing a dosage form of antimicrobial material (preferably natamycin) comprising (i) co-processing antimicrobial material (preferably natamycin) with a physiologically acceptable encapsulating material to cause the material to encapsulate the antimicrobial material (preferably natamycin) inside a breastplate, and (ii) recovering a product of protected food preservative antimicrobial material (preferably natamycin). 18. A method according to paragraph 17, wherein said encapsulation process is selected from a fluidized bed process, liposome encapsulation, spray drying, spray cooling, extrusion, co-extrusion, coacervation and mixtures thereof. . 19. A method according to paragraph 17, wherein the encapsulating material comprises a material selected from the group consisting of hydrophobic materials, hydrocolloid materials and mixtures or combinations thereof. 20. A process according to paragraph 17, wherein the encapsulation process comprises fluidized bed encapsulation of antimicrobial material (preferably natamycin) with an encapsulating material in an aqueous solution or suspension or in a molten state. 21. A method according to paragraph 17, wherein the encapsulation process comprises coacervation of antimicrobial material (preferably natamycin) with an encapsulating material. 22. A process according to paragraph 19, wherein the encapsulating material comprises a hydrocolloid or a mixture of hydrocolloids. 23. A method according to paragraph 17, which includes the steps of a) providing an aqueous phase and antimicrobial material (preferably natamycin) incorporated in the aqueous phase, b) providing a hydrophobic phase in a molten form, c) incorporating or dissolving an encapsulating material or mixture of encapsulating materials in the aqueous phase or in the hydrophobic phase d) combining the aqueous phase with the hydrophobic phase and homogenizing or mixing the combined phases to form a water-in-oil emulsion, e) encapsulating the aqueous phase in the emulsion, whereby a dispersion comprising encapsulated aqueous spheres is formed and the antimicrobial material (preferably natamycin) is encapsulated in the aqueous spheres, and f) processing the dispersion obtained in step e) to form microcapsules wherein the aqueous spheres encapsulated are additionally encapsulated in solidified hydrophobic shell material. 24. A method for preserving a food product comprising adding to the food product a preservative amount of effective food of antimicrobial material (preferably natamycin) which is encapsulated within a physiologically acceptable shell.
25. A method according to paragraph 24, wherein the encapsulated antimicrobial material (preferably natamycin) is added to the food product before or in connection with the production of the food product and said shell is effective to protect the encapsulated antimicrobial material (preferably natamycin) against degradation by conditions used in the production or storage of the food product, said shell providing the release of antimicrobial material (preferably natamycin) in the food product. 27. A method according to paragraph 24, wherein the food product is selected from a dressing for salads, a seasoning, a tomato sauce, a mash, a hot sauce, a pickled gherkin, a spreadable sauce for snacks, an acidic dairy product including natural cheese, cottage cheese, acidified cheese, cream cheese, yogurt, sour cream and processed cheese, a fruit juice, an acidic beverage, an alcoholic beverage including wine and beer, a chilled dough, a pastry product cooked or uncooked, a dairy filling or cover, a top cover or coating, a marinade, marinated or breaded meat or poultry, a pizza top or base, a quick food product, a snack or food kit, a equipment to make a confectionery product, combinations thereof, pet food and roasting food. 28. A method according to paragraph 24, wherein the encapsulated antimicrobial material (preferably natamycin) is included in a dough for a yeast high or non-yeast high pastry product. 29. A method of conformance with paragraph 28, wherein the dough is baked in bread and subsequently sliced. 30. A preserved food product comprising as preservative an effective food preservation amount of antimicrobial material (preferably natamycin) that is encapsulated within a physiologically acceptable shell. 31. A food product according to paragraph 30, wherein the food product is selected from a dressing for salads, a seasoning, a tomato sauce, a puree, a hot sauce, a pickled gherkin, a spreadable sauce for snacks , an acidic dairy product including natural cheese, cottage cheese, acidified cheese, cream cheese, yogurt, sour cream and processed cheese, a fruit juice, an acidic beverage, an alcoholic beverage, a chilled dough, a baked goods product or not cooked, a milk filling or cover, a top cover, a marinade, marinated meat or poultry, breaded meat or poultry, a pizza cover or base, a quick food product, a snack or food kit, a equipment to make a confectionery product, combinations thereof, pet food and roasting food. 32. A food product according to paragraph 31, wherein the confectionery product is sliced or sliced bread.
Highly Preferred Aspects Some highly preferred aspects of the present invention are discussed below: a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nisin core and shell of encapsulating material, and (ii) introducing encapsulated nisin into or onto the food by (a) injecting the nisin encapsulated in the feed or (b) stirring the encapsulated nisin with the feed. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (i) introducing encapsulated nisin into or on the food at (a) ) injecting the encapsulated nisin into the feed or (b) stirring the encapsulated nisin with the feed, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject the encapsulated nisin in the feed or (b) stir the nisin encapsulated with the feed, wherein the encapsulated nisin has an average particle size less than 150 μ? t ?. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject encapsulated nisin into the food or (b) stir the nisin encapsulated with the food, wherein the shell is or comprises selected material of triglyceride and carrageenan. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject the encapsulated nisin in the food or (b) stir the encapsulated nisin with the feed, where the feed is raw meat. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject the encapsulated nisin into the feed or (b) stir the encapsulated nisin with the feed, wherein the encapsulated nisin is introduced into or onto the feed in a brine vehicle. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting the nisin encapsulated in the food or (b) stirring the nisin encapsulated with the food, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the encapsulated nisin has an average particle size of less than 150 μ. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting the nisin encapsulated in the food or (b) stirring the nisin encapsulated with the food, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the shell is or comprises selected material of triglyceride and carrageenan. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting the nisin encapsulated in the food or (b) stirring the nisin encapsulated with the food, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, where the food is raw meat. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting the nisin encapsulated in the food or (b) stirring the nisin encapsulated with the food, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the encapsulated nisin is introduced into or onto the food in a brine vehicle. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject encapsulated nisin into the feed or (b) stir the nisin encapsulated with the feed, wherein the encapsulated nisin has an average particle size of less than 150 μ ??, wherein the shell is or comprises selected triglyceride material and carrageenan. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject encapsulated nisin into the feed or (b) stir the encapsulated nisin with the feed, wherein the encapsulated nisin has an average particle size less than 150 μ, where the feed is raw meat. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject the encapsulated nisin into the feed or (b) stir the encapsulated nisin with the feed, wherein the encapsulated nisin has an average particle size of less than 150 [mu], wherein the encapsulated nisin is introduced into or onto the feed in a brine vehicle. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject encapsulated nisin into the food or (b) stir the nisin encapsulated with the food, wherein the shell is or comprises selected material of triglyceride and carrageenan, wherein the feed is raw meat. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting encapsulated nisin into the feed or (b) stirring the nisin encapsulated with the feed, wherein the shell is or comprises selected material of triglyceride and carrageenan, wherein the encapsulated nisin is introduced into or onto the feed in a carrier vehicle. brine. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject the encapsulated nisin into the feed or (b) stir the encapsulated nisin with the feed, wherein the feed is raw meat, wherein the encapsulated nisin is introduced into or onto the feed in a brine vehicle. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nisin core and shell of encapsulating material, and (ii) introducing encapsulated nisin into or onto the food by (a) injecting the nisin encapsulated in the feed or (b) stirring the encapsulated nisin with the feed, wherein the nisin is present in an amount to provide a microbicidal effect or microbiostatic with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the feed is raw meat, wherein the encapsulated nisin has an average particle size less than 150, μ? t ?. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting the nisin encapsulated in the food or (b) stirring the nisin encapsulated with the food, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, where the food is raw meat, where the shell is or comprises selected material of triglyceride and carrageenan. . · A method for introducing an antimicrobial material into a food comprising "(i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) ) injecting the encapsulated nisin into the feed or (b) stirring the encapsulated nisin with the feed, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Camobacterium, Enterococcus Usteria monocytogenes, Bacillus, Ciostrium, and Brochothrix thermosphacta, where the food is raw meat, where the encapsulated nisin is introduced into or onto the food in a brine vehicle • a procedure to introduce an antimicrobial material into a food that comprises (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of material in capsulant, and (ii) introducing encapsulated nisin into or on the food by (a) injecting the nisin encapsulated in the feed or (b) stirring the encapsulated nisin with the feed, wherein the nisin is present in an amount to provide an effect microbicidal or microbiostatic with respect to a microorganism selected from Lactobacillus, Leuconostoc, Camobacterium, Enterococcus; Usteria monocytogenes, Bacillus, Ciostrium; and Brochothríx thermosphacta, wherein the encapsulated nisin is introduced into or onto the feed in a brine vehicle, wherein the encapsulated nisin has an average particle size of less than 150 μ. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) injecting the nisin encapsulated in the food or (b) stirring the nisin encapsulated with the food, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the encapsulated nisin is introduced into or onto the food in a brine vehicle, wherein the shell is or comprises selected material of triglyceride and carrageenan. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject encapsulated nisin into the food or (b) stir the encapsulated nisin with the feed, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the encapsulated nisin is introduced into or onto the food in a brine vehicle, where the food is raw meat. • a method for introducing an antimicrobial material into a food comprising (i) providing nisin in an encapsulated form comprising a nucleus of nisin and shell of encapsulating material, and (ii) introducing encapsulated nisin into or on the food at (a) inject encapsulated nisin into the food or (b) stir the encapsulated nisin with the feed, wherein the nisin is present in an amount to provide a microbial or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Camobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the encapsulated nisin has an average particle size less than 150 μ ??, wherein the shell is or comprises selected material of triglyceride and carrageenan, wherein the feed is raw meat, wherein the encapsulated nisin is introduced into or onto the food in a brine vehicle. "An antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin; an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Camobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta. · An antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the encapsulated nisin has an average particle size of less than 150 μ? T ?
• an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the shell is or comprises selected material of triglyceride and carrageenan.
• an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with with respect to a microorganism selected from Lactobacillus, Leuconostoc, Camobacterium, Enterococcus; Listeria monocyfogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the encapsulated nisin has an average particle size less than 150 μ ??). • an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with with respect to a microorganism selected from Lactobacillus, Leuconostoc, Camobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothrix thermosphacta, wherein the shell is or comprises selected material of triglyceride and carrageenan. • an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the encapsulated nisin has an average particle size of less than 150 μ? , wherein the shell is or comprises selected material of triglyceride and carrageenan. • an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothríx thermosphacta, wherein the feed is raw meat, wherein the encapsulated nisin has an average particle size of less than 150 μ. • an antimicrobial material in an encapsulated form, comprising a nucleus of nisin and shell of encapsulating material, wherein the shell of encapsulating material is impermeable to nisin, wherein the nisin is present in an amount to provide a microbicidal or microbiostatic effect with respect to a microorganism selected from Lactobacillus, Leuconostoc, Carnobacterium, Enterococcus; Listeria monocytogenes, Bacillus, Clostridium; and Brochothríx thermosphacta, where the food is raw meat, where the shell is or comprises material selected from triglyceride and carrageenan.
The present invention will now be described in greater detail in the following examples.
EXAMPLES
Nisina
EXAMPLE 1
First, a solution of 15 g of k-carrageenan in 1000 ml of phosphate pH regulator at pH 3.5 is prepared at 85 ° C. To this is added 300 g of Nisaplin® (Danisco's commercial nisin extract: equivalent to 1 x 106 lU / g of nisin potency). The resulting mixture is mixed uniformly. At the same time, a mixture of 1333 g of a vegetable triglyceride (Danisco: GRINSTED®, PS 101, mp 58 ° C) and 73 g of acetylated emulsifier (Danisco: Acetem 50 00) melts at 85 ° C in a bath Water. The melted fat mixture is kept under homogenization (Silverson mixer, 8 kRPM) as the aqueous mixture is slowly incorporated. The homogenization is maintained for 5 minutes after the entire aqueous mixture is added and then a solution of 3 g of polysorbate 80 in 40 ml of water is added under constant mixing. The resultant low viscosity water-in-oil emulsion is then spray-cooled immediately in a Niro spray tower using the following parameters: inlet air temperature: 10 ° C, outlet air temperature 28 ° C, rotating atomizing wheel speed: 10 kPRM. A free-flowing powder is obtained. The encapsulated nisin can be used for injection or stirring of raw meat which is then cooked immediately. The release of nisin from the fat shell would occur under injection and / or cooking. Since the encapsulated fat-based shell material would cause the particles to float to the surface of the brine and injection, either a) a viscosity agent such as xanthan could be used to stabilize the particles in the brine, or b) to Mix the brine before using it as an injection material. The mixing of the particles would occur naturally when the encapsulated nisin is used in the brine used to stir the meat. The same encapsulated material could be used for sustained release at cooled temperature of encapsulated nisin within marinates used in raw, cooled, vacuum packed meat.
EXAMPLE 2
First, a solution of 15 g of sodium alginate in 1000 ml of phosphate pH regulator at pH 3.5 is prepared at 85 ° C. To this is added 300 g of Nisaplin® (Danisco's commercial nisin extract: equivalent to 1 x 106 lU / g of nisin potency). The resulting mixture is mixed uniformly. At the same time, a mixture of 1333 g of a triglyceride vegeta) (Danisco: GRINSTED®, PS 101, melting point 58 ° C) and 73 g of acetylated emulsifier (Danisco: Acetem 50 00) melts at 85 ° C in a water bath. The melted fat mixture is kept under homogenization (Silverson mixer, 8 kRPM) as the aqueous mixture is slowly incorporated. After the addition of the aqueous mixture, a solution of 7 g of calcium chloride in 70 ml of water is added dropwise. The homogenization is maintained for another 5 minutes and then a solution of 3 g of polysorbate 80 in 40 ml of water is added under constant mixing. The resulting low viscosity water-in-oil emulsion is then spray-cooled immediately in a Niro spray tower using the following parameters: inlet air temperature: 10 ° C, outlet air temperature 28 ° C, air wheel speed Rotating atomization: 10 kPRM. A free-flowing powder is obtained. The use of this encapsulated nisin is as described in example 1.
EXAMPLE 3
A solution of 1 g of bilayer-forming lipid and 100 mg of cholesterol in a suitable organic solvent is evaporated to form a thin dry lipid film at the bottom of the container. After uniform drying of the lipid film, 1 l of nisin-containing water (such as Nisaplin) at the saturation concentration is added to the container and the mixture is uniformly mixed or homogenized. The resulting multilamellar vesicle suspension (MLV) can be further processed by microfluidization to form a smaller, more homogeneous unilamellar vesicle (SUV). The nisin suspension encapsulated in liposome can be added directly to the meat by injection / stirring. These particles are small enough to pass through injection needles without disintegration of the liposome cuirass. The liposome-encapsulated nisin would be released during cooking since the liposomes are broken at 45-50 ° C due to the transition temperature of the bilayer-forming phospholipid / amphiphilic compounds. The nisin encapsulated in liposome would be released slowly over time, making it suitable for sustained release in marinated raw meat. Nisin encapsulated in liposome could be made by several procedures, including microfluidization, extrusion, 'French press', reverse phase evaporation, freeze-thaw cycle, etc. Microfluidization is the preferred aspect since it is a continuous, high-capacity, solvent-free process.
EXAMPLE 4
The use of a fluidized bed to apply a hydrophobic shell on the nisin. If the particle size of nisin is too fine, the powder can be agglomerated in a suitable equipment using a binder solution (solution of sticky hydrocolloids such as alginate or maltodextrin) in order to obtain a dense powder of particle size between 100- 150 micrometers The appropriate powder is then introduced into the coating chamber of a fluidized bed microencapsulation unit and is fluidized at an inlet air flow rate of 5-30 cm / s and at a temperature up to 50 ° C to fluidize the particles. A coating material is then sprayed onto the fluidized bed of antimicrobial material using a double fluid nozzle and high pressure atomizing air. In one example, a molten mixture of triglyceride and additives is sprayed onto the nisin to form a continuous layer of fat around each individual particle as the melted fat spreads and solidifies on the particles. The amount of applied grease can be up to 50%, but generally not less than 20% p / p. In another example, a dispersion of coating material in water or a solution of coating material in ethanol is sprayed onto the fluidized particles and the fluidizing air is used to evaporate the solvent or water, which leaves behind a continuous film of material of coating on the antimicrobial particles. Examples of coating material in this case include lacquer, zein or any other hydrophobic coating materials. For the encapsulated nisin prepared by this method to be used for injection of raw meat, the particle size must be less than 175 microns. In addition, the particle size must be greater than 100 microns for the fluidization process to work.
EXAMPLE 5 Improved antilisteriai effect with nisin encapsulated in sausages
The encapsulated nisin is prepared by crystallization by spraying in accordance with the following procedure. Fully hydrogenated triglyceride (GRINSTED PS101,100 parts) is melted at 85 ° C in a water bath. The nisin (64 parts) is pre-heated to 50 ° C and added to the molten triglyceride, maintained at 85 ° C, under vigorous mixing. The mixing is maintained until the mixture becomes smooth and free of lumps. The suspension is then pumped to the atomization device of a spray tower in tracked tubes maintained at 75-85 ° C. The atomization device is a "rotating wheel" at 9000 RPM installed on the top of the spray tower. The cooled air (3-5 ° C) is blown into the spray tower to crystallize the atomised grease / nisin droplets before reaching the tower walls. The solidified powder is collected at the bottom of the tower. The powder can be maintained at 40 ° C 2-3 days to allow the crystallization of the fat phase, if necessary, from the alpha form to the beta form. The anti-cake agent, such as calcium stearate or silicon dioxide can be added at a level of 0.1-1% to avoid forming additional groups of the powder. An inoculation assay with monocytogenes üstería was conducted with sausages; this demonstrated the thermoprotective benefit of the encapsulated nisin. The formulation of the sausages was as follows (raw batch weight basis): 74.1% of cuts of meat (lean beef and pork fat), 1.66% of NaCl, 1.48% of corn syrup solids, 0.74% of HMP, 0.37% abstention of hydrolyzed, 0.33% of sodium thpoliphosphate, 0.37% of spice / seasoning mixture, 0.037% of erythorbate, 0.185 of sodium nitrite cure mixture, 13.3% of added water, 7.4% of water added (10%, shrinkage by cooking). Nisin was added at 250 and 500 IU / g either as encapsulated nisin (Nisaptin®, Danisco) or as an encapsulated nidin product. The sausage, which contained 28% fat, was then passed through a heating / smoking regime as shown below:
Smoked appliance program
The sausages were maintained at an internal end point temperature of 71.1 ° C, cooled with a shower at 35.5 ° C, then cooled to < 9.4 ° C. The sausages were packed under vacuum, six to one bag and inoculated on the surface with a mixture of 5 strains of Listeria monocytogenes and Listeria innocua (including environmental isolates). The nisin levels in the sausages were measured the next day and during storage for 12 weeks at 3.3-4.4 ° C by a bioassay method, a horizontal agar diffusion assay method using Micrococcus luteus as the indicator organism (Fowler et al. 1975. Society for Applied Bacteriology Technical Series 8: 91-105). This uses an acid / heat extraction step that detects all the residual nisin within the samples, even if they are encapsulated. The sausages were also analyzed at weekly intervals for counts of Listeria monocytogenes and lactic acid bacteria natural contaminants. The long heat processing resulted in significant nisin loss. The initial nisin levels detected in the sausages after processing were much higher in the samples to which encapsulated nisin had been added compared to those with non-encapsulated nisin (Nisaplin®, Danisco) (see Figure 1). The microbiological data of the trial were subjected to multivariate statistical analyzes. This concluded that the encapsulation achieved a greater initial drop in Listeria numbers. The optimum treatment to achieve a shelf life of 84 days was provided by encapsulated nisin (at 500 lU / g), the optimal secondary treatment was Nisaplin® (at 500 lU / g). This also demonstrated the superior efficacy of the encapsulated nisin.
EXAMPLE 6 Enhanced nisin levels with nisin encapsulated in a confectionery product
Soft buns are confectionery products based on high moisture flour that have been implicated in outbreaks of food poisoning due to Bacillus cereus. The products are stored at room temperature and during the shelf life of 5 days, heat resistant spores of Bacillus cereus (present in the flour) can germinate and grow, particularly in countries with warm climates. Nisin has been used as a preservative in soft buns to prevent the growth of this pathogen and to guarantee consumer safety. The baking procedure for soft buns, however, can result in significant nisin loss. This involves heating a hot plate for 3-5 minutes. The test described below demonstrated the thermo-protective effect of encapsulation, ensuring that a higher percentage of nisin added survived the baking process. The soft buns were prepared by a normal production method, with the addition of nisin (such as Nisaplin®) or encapsulated nisin (prepared by spray crystallization according to the procedure of example 5) to the dough before cooking on the plate hot. The soft buns (pH 5.6-6.0, water activity 0.8-0.9) were then incubated at room temperature (21 ° C) for 5 days. The nisin levels in the soft buns were measured the next day by a bioassay method (see above).
Test samples 1. Nisaplin® (Danisco). Nisin potency 1 x 106 lU / g 2. Encapsulated nisin sample NAP 03228 (Danisco). Power of nisina 5.36 x 105
Soft buns test results
The average residual nisin levels that resulted from the encapsulation were 61% compared to the average residual nisin levels of 22% for non-encapsulated nisin.
EXAMPLE 7 Improved nisin levels in processed cheese
The encapsulated nisin (prepared by spray crystallization according to the procedure of Example 5) and the unencapsulated nisin (Nisaplin®, Danisco) were added to a commercial processed cheese formulation, after which processed cheese samples were subjected to 10 minute heating step at core temperatures of 60 ° C, 80CC and 100 ° C. After heating, the residual nisin levels were measured in the processed cheese, using heat / acid extraction and the horizontal agar diffusion method.
The results show higher nisin levels after heat treatment for encapsulated nisin samples compared to non-encapsulated nisin samples.
EXAMPLE 8 Improved nisin levels and efficacy in a pasta meat sauce due to encapsulation protection
The encapsulated nisin (prepared by spray crystallization according to the procedure of Example 5) and encapsulated nisin (Nisaplin®, Danisco) were added to a Bolognese sauce, pH 5.69. This was prepared according to the following recipe: ground lean meat (50.0 g), tomatoes lightened in tomato juice (48.9 g), starch (0.5 g), salt 0.4 g, sugar (0.2 g). The ground beef was fried for 5 minutes until golden brown. The dry ingredients were mixed in the meat and then the tomatoes were added. The sauce was simmered for 10 minutes and then allowed to cool. A blender was used to produce a mild sauce to facilitate sampling. The sauce was diluted to facilitate the test and the pH was adjusted. Additions of nisin preparations based on equivalent nisin potencies were made at a level of 250 lU / g to the sauce, which was then pasteurized at a core temperature of 80 ° C for 30 minutes. The samples were then inoculated with a combination of strains of Listeria monocytogenes (strain 272, CRA3930, 358, NCTC12426) at 102 CFU / g. The nisin levels were immediately tested using the horizontal agar diffusion method. The samples were stored at 8 ° C and analyzed microbiologically at regular intervals.
The results show that a high percentage of nisin survived the procedure with heat if it was encapsulated and this had the result of better control of Usteria in the food.
EXAMPLE 9 Improved nisin levels in a raw meat matrix
To test the protective effect of encapsulation against degradation or inactivation of nisin in raw meat, samples of encapsulated and non-encapsulated nisin (as in example 8) were added to the raw minced meat. Supply solutions of the nisin preparations were prepared in 0.01 M HCL, which was added to the meat. All nisin additions were equivalent, based on 200 IU / g. After incubation overnight at 4 and 20 ° C, the nisin levels were analyzed by the horizontal well diffusion assay method (same as before). Nisin preparation Nisin detected after overnight incubation (IU / g) 4 ° C 20 ° C Non-encapsulated Nisin (A) 120 60 Encapsulated Nisin (C) 169 136 Encapsulated Nisin (D) 157 124 The results show that the encapsulation helped protect nisin against inactivation or degradation in raw meat. After incubation overnight at 20 ° C, encapsulated nisin levels dropped to 60 lU / g compared to 124-136 lU / g if encapsulated.
Natamycin
EXAMPLE 0 Production of encapsulated natamycin by a coacervation procedure
First, a solution of gelatin (219 g, isoelectric point = 8) in 6 liters of water at 50 ° C was prepared. Second, a solution of 219 g of acacia gum was dissolved in 6 liters of water at 50 ° C. The two solutions were mixed together and kept at 45 ° C under vigorous stirring. 700 g of Natamax ™ SF (Danisco) were added to the aqueous solutions and the pH was rapidly reduced to 4.0 using 1M HCL, after which the temperature was reduced to 5 ° C at the rate of about 1 ° C / min, keeping the excitement all the time. 36 ml of a 1: 1 aqueous solution of glutaraldehyde was added, the pH was re-adjusted to 8.5 using aqueous 1M NaOH and then the temperature was again increased to 45 ° C at a rate of about 2 ° C / min. Finally, the entire mixture was spray-dried in a spray tower using a double fluid nozzle mounted in the source configuration, an inlet temperature of 180 ° C and a spray rate to maintain the outlet air temperature at approximately 100 ° C. In an alternative procedure, 1 kg of each of the gum arabic and maltodextrin (DE 12) are dissolved in the aqueous mixture immediately before spray drying.
EXAMPLE 11 Encapsulation in fluid bed of natamycin
Preprocessing If the particle size of natamycin is too fine (below 100 microns average) the powder agglomerates to a larger average particle size for easier processing by fluidized bed. The larger average particle size not only makes the procedure easier, but also allows the use of less coating material while achieving the same protection as with more shell material. The natamycin is agglomerated in a suitable equipment such as a high shear mixer (such as a Lodige mixer) using a binder solution (solution of sticky hydrocolloids such as alginate or maltodextrin) in order to obtain a dense powder of particle size above 150, preferably between 200-350 μ? t? and volumetric density above 0.4, preferably 0.7 g / cm3.
Encapsulation in a fluidized bed under hot melt 3 kg of agglomerated natamycin is introduced into the coating chamber of an Aeromatic-Fielder P1 fluidized bed microencapsulation unit and is fluidized using an inlet air flow rate of 80 cm / s and temperature of 43 ° C. A molten hydrogenated triglyceride maintained at 85 ° C is then separated in the fluidized bed of antimicrobial using a peristaltic pump and a double fluid nozzle set at 2 bar and 2 m 3 air / hr. The grease is applied at about 1 kg / hr, so as to form a continuous layer of fat around each individual particle as the molten fat disperses and solidifies on the particles. Sufficient fat is applied to reach a final product that contains 30% fat and 70% natamycin.
EXAMPLE 12 Encapsulation by extrusion of natamycin
A mixture of 60 parts of corn starch, 25 parts of natamycin and 10 parts of polyethylene glycol and 5 parts of water are mixed together and the first cylinder heated to 40 ° C is introduced into a twin screw extruder. The mass is treated at 100 ° C for just a few seconds in cylinders 2 and 3 then cooled to 45 ° C in the cylinders to the die. Alternatively, a vacuum pump is installed in the last cylinder to draw water. The tape that comes out is cut into pieces of between 250 and 500 μ.
EXAMPLE 13 Use of natamycin encapsulated in orange juice
Natamycin was encapsulated by a coacervation method as described in example 10, using either gelatin and acacia as a shell material (NAP03015), or gelatin, acacia and maltodextrin (NAP03023). The samples, together with natamycin such as Natamax ™ (Danisco) were added to orange juice (pH 3.85) and heated at 100 ° C for 10 minutes. The residual natamycin levels in the juice before and after treatment were tested by HPLC. The samples were diluted in methanol for this test. The results are shown in table 1. The experiment shows that the microcapsule prevented the release of natamycin, so not all of the estimated natamycin present could be detected before the passage of heating. After heating, encapsulated natamycin showed higher recovery levels than with unprotected natamycin.
TABLE 1 Heat protection of natamycin encapsulated in orange juice
EXAMPLE 14 Use of natamycin encapsulated in vinaigrette
A dressing of vinaigrette was prepared which contained water (494.6 ml), 10% vinegar (220 ml), sugar (90 g) and salt (10 g), pH 2.6. Additions of encapsulated and unencapsulated natamycin were made as shown in table 2. Sample NAP03015 was encapsulated by coacervation as described in example 10. Sample NAP03007 was encapsulated by spray cooling with a triglyceride shell material.
TABLE 2
The vinaigrette was incubated at 25 ° C, and the samples were tested for residual natamycin content at regular intervals. The vinaigrette was shaken before each sample, and a sample taken for 14PLC analysis, which was diluted 1: 1 in methanol. The levels of natamycin found in the mixed vinaigrette and in the water layer are only shown in tables 3 and 4. The results show that the encapsulation protects natamycin against acid degradation in the vinaigrette, which allows a slow release of the conservative with time. Sample NAP03007 contained only a small amount of unencapsulated natamycin at the beginning of the experiment.
TABLE 3 Natamycin detectable in a vinaigrette dressing at 25 ° C (sample taken from homogenized dressing) Days at 25 ° C Percentage of natamycin estimated addition level (based on estimated addition level) Natamax NAP03007 NAP03015 0 70.5% 1.8% 70 % 1 38% 4.5% 50.5% 6 22.5% 19.3% 23.8% 9 13% 29.5% 36.5% 14 10% 40.8% 29% 21 4.5% 17.5% 10.2% TABLE Natamycin detectable from the water phase of a dressing of vinaigrette a 25 ° C
EXAMPLE 15 Use of natamycin encapsulated in bread
A bread is made by preparing a dough that contains flour, water, yeast, salt and a dough conditioner. The mass mixture includes either natamycin or encapsulated natamycin or none at all. Both natamycin preparations were added at a potency dose of 12 ppm (0.0012%) on a weight of flour and these were added together with the other dry ingredients. All the dry ingredients are mixed together evenly between 3 and 10 minutes. The dough is then given a short rest period after mixing (about 5 to 10 minutes) followed by scaling but required. A second rest period is then applied following a second molding for the shape of the dough as desired. The dough is then placed on a sheet or tray. A lifting period of approximately 50 minutes at 85% relative humidity at 4 ° C is given below. The fully tested dough is then baked at 190 to 230 ° C for approximately 15 to 30 minutes. Bread containing unencapsulated natamycin shows poor elevation, while the elevation of the encapsulated natamycin proceeds in a manner similar to the control bread that does not contain any natamycin. This demonstrates the benefit of encapsulation, which prevents natamycin from inhibiting the fermentation reaction of the yeast. When the bread cools, the content of natamycin in the bread is tested. The natamycin content of the bread containing encapsulated natamycin is greater than the bread containing natamycin non-encapsulated, indicating the thermoprotective benefit of the encapsulated natamycin. The bread is then sliced and observed during the normal shelf life period for mold growth. The delay of putrefaction of molds is observed for natamycin containing bread. This shelf life extension is greater for bread containing encapsulated natamycin, which is a reflection of the higher natamycin levels that survive the baking process.
EXAMPLE 16 Encapsulation of natamycin in a double breastplate
First, a solution of 15 g of kappa-carrageenan in 1000 ml of phosphate pH regulator at pH 7.0 is prepared at 85 ° C. To this is added 300 g of commercial natamycin (Natamax ™ SF, Danisco). The resulting mixture is mixed uniformly. At the same time, a mixture of 1333 g of vegetable triglyceride (GRINDSTED ® PS 101, p.p. 58 ° C) and 73 g of acetylated emulsifier (Acetem 50 00) is melted at 85 ° C in a water bath. The melted fat mixture is kept under homogenization (Silverson mixer, 8000 rpm) as the aqueous mixture is slowly incorporated. The homogenization is maintained for 5 minutes after the entire aqueous mixture is added and then a solution of 3 g of polysorbate 80 in 40 ml of water is added under constant mixing. The resulting low viscosity water-in-oil emulsion is spray-cooled immediately afterwards in a Niro spray tower using the following parameters: inlet air temperature 10 ° C, outlet air temperature 28 ° C, atomization wheel speed rotating 10 000 rpm. A free-flowing powder is obtained. The incorporation of natamycin encapsulated in an orange juice results in a much more stable natamycin formulation compared to using natamycin not encapsulated in the liquid, thus drastically improving the survival rate of natamycin in the beverage. The encapsulated natamycin, as presented in this example, is released at a rate of only 7% after three days.
References
• Background Davies, E. A., Bevis, H. E., Potter, R., Harris, J., Williams, G. C. and Delves-Broughton, J. 1998. The effect of pH on the stabiiity of nisin solutions during autoclaving. Letters n Applied Microbiology 27: 186-187. • De Vuyst, L, and Vandamme, E. J. 1994. Nisin, a lantibiotic produced by Lactococcus lactis subsp lactis: properties, biosynthesis, fermentation and applications. In Bacteriocins of Lactic Acid Bacteria. Microbiology, Genetics and Applications, eds. L. de Vuyst and E. J. Vandamme pp 151-221. London: Blackie Academic and Professional. • Rose, N. L, Palcic, M. M., Sporns, P. and McMullen. 2002. Nisin: a novel substrate for glutathione S-transferase isolated from fresh beef. Journal of Food Safety 67: 2288-2293. • Rose, N. L, Sporns, P., Stiles, M. E., and McMullen, L 1999. Inactivation of nisin by glutathione in fresh meat. J. Food Science 64: 759-762.
• Rose, N. L, Sporns, P., Dodd, H.M., Gasson, M.J., Mellon, F.A., and McMullen, L 2003. Involvement of dehydroalanine and dehydrobutyrine in the addition of glutathione to nisin. J. Agrie Food chem. 51: 3174-3178. • Susiluoto, T., Korkeala, H., and Bjorkroth, K. J. 2003. Leuconstoc gasicomitatum is the dominating lactic acid bacterium in retail modified atmosphere packaged marinated broiler meat strips on sale by day. International Journal of Food Microbiology 80: 89-97.
• Thomas, L.V., Clarkson, M.R., and Delves-Broughton, J. 2000. Nisin. In: Natural Food Antimicrobial Systems. Ed. A. S. Naidu. Pp 463-524. E.U.A: CRC Press. • Vamam, A. H., and Sutherland, J. P. 1995. Meat and Meat Products. Technology, Chemistry and Microbiology. Chapman & Hall. London
• Axelsen, L. 1998. Lactic acid bacteria: classification and physiology. In: Salminen, S. and von Wright, A. In: Lactic Acid Bacteria. 2nd Ed. New York, Marcel Dekker, pp 1-72. • Delves-Broughton, J. 1990. Nisin and its uses as a food preservative. Food Technol. 44: 100, 102, 104, 106, 108, 111-112, 117. • Hoover, D.G. 1993. Bacteriocins with potential for use in foods. In: Antimicrobials n Foods. Ed: P.M. Davidson and A. L. Branen. Marcel Dekker, E.U.A. • Hurst, A. 1981. Nisin. Adv. Appl. Microbiol. 27: 85-123 · Hurst, A. 1983. Nisin and other inhibitory substances from lactic acid bacteria. In Antimicrobials in Foods. Eds. A. L. Branen and P. M. Davidson, pp. 327-351. New York: Marcel Dekker. • Naidu, A. S. (Ed.) 2000. Natural Food Antimicrobial Systems. E.U.A: CRC Press. · Ray, B., and Miller, K. W. 2003. Bacteriocins other than nisin: the pediocin-like cysibiotics of lactic acid bacteria. In: Natural Antimicrobials for the Minimal Processing of Foods. Ed: Sibel Roller. CRC Press, E.U.A. • Ray, B. and Daeschel, M. A. 1994. Bacteriocins of starter culture bacteria. In: Natural Antimicrobial Ssytems and Food Preservation. 1994. Ed: Dillon, V. M. and Board, R. G. CAB International, R.U., pp. 133-166. • Ray, B., Miller, K. W. and Jain, M. K. 2001. Bacteriocins of lactic acid bacteria. Indian Journal of Microbiology 41: 1-21. • Thomas, L. V., and Delves-Broughton, J. 2001. New advances in the application of the food preservative nisin. Research Advances in Food Science 2: 1-22 • Wessels, S., Jelle, B., and Nes, I. F. 1998. Bacteriocins of the Lactic Acid Bacteria: An Overlooked Benefit for Food. Danish Toxicology Center, Denmark.
Invention of nisin in meat • Caserío, G., Ciampella, A., Gennari, M., and Barluzzi, A.M. 1979. Industrie Alimentan 18: 1-12. Research on the use of nisin in cooked, cured meat products. • Gola, J. 962. 'Preservation of canned hams stored at unusual temperatures'. Collected Reports of Research Institute for Meat (Brno) 10: 239-244. • Taylor and Somers. 1985. Evaluation of the antibotulinal effectiveness of nisin in bacon. Journal of Food Protection 48: 949-952. • Usbome, WR, Collins-Thompson, DL and Wood, DS. 1986. Sensory evaluation of nisin-treated bacon. Dog. Inst. Food Sci. Technol. J. 19: 38-40.
• US 2003/0108648 A1 2003 (Rhodia) 'Composition having bacteriostatic and bactericidal! activity against bacterial spores and vegetative cells and process for treating foods therewith '. Ming, King and Payne. • US 6207210 B1. (Rhodia). 'Broad-range antibacterial composition and process of applying to food surfaces. Bender, King, Ming and Weber.
Patent filed on March 27, 2001. • EP 0 770336 A1. European patent application. 1995. Nestle. Process for preparing a meat product. • Internet article [http://www.nal.usda.gov/fsrio/ppd/ars010f.htm] about work in Meat
Research Unit, MARC mentions a presentation on 'antibacterial properties of injectable beef marinades'. This seems to be directed to E. coli 0157 and it would be unlikely that it was nisin.
Encapsulation of nisin • Benech, R. -O, Kheadr, EE, Laridi, R., and Fliss, I. 2002. Inhibition of Listeria innocua cheddar cheese by addition of nisin Z in liposomes or by in situ production in mixed culture . Applied & Environmental Microbiology 68: 3683-3690. · Bower, C.K., McGuire, J. and Daeschel, M.A. 1995. Influences on the antimicrobial activity of surface-adsorbed nisin. J. Industrial Microbiology 15: 227-233. • Bower, C.K., McGuire, J. and Daeschel, M.A. 1995. Suppression of Listeria monocytogenges colonization following adsorption of nisin onto silica surfaces. Appl. Environ. Microbe! 61: 992-997. • Cahill, S.M., Upton, M.E., and McLoughlin, A.J. 2001. Bioencapsulation technology in meat preservation. In: Applied Microbiology. Eds Durieux, A., and Simón, J. P. Dordrecht: Kluwer Academic Publishers. pp 239-266. • Cutter, C. N., and Siragusa, G. R. 1996. Reduction of Brochothrix thermosphacta on beef surfaces following immobilization of nisin in calcium gels. Letts. Applied Microbiology 23: 9-12. • Cutter, C. N., and Siragusa, G. R. 1997. Growth of Brochothrix thermosphacta in ground beef following treatments with nisin in calcium alginate gels. Food Microbiol 14: 425-430. • Cutter, C. N., and Siragusa, G. R. 1998. Incorporated of nisin into meat binding system to inhibit bacteria on beef surfaces. Letts. Applied Microbiol. 27: 19-23. • Daeschel, M.A., McGuire, J., and Al-Makhlafi, H. 1992. Antimicrobial activity of nisin adsorbed to hydrophilic and hydrophobic silicon surfaces. J. Food Protection 55: 731-735. • Degnan, A. J., and J. B. Luchansky. 1992. Influence of beef tallow and muscle on the antilisterial activity of pediocin AcH and liposome-encapsulated pediocin AcH. J. Food Protection 55: 552-554. • Degnan, A.J., Buyong, N., and Luchansky, J. B. 1993. Antilisterial activity of pediatric AcH in model food systems in the presence of an emulsifier or encapsulated within liposoms. International Journal of Food Microbiology 18: 127-138. • Laridi, R., Benech, R. -O, Vuillemard, J. C, Lacroix, C, Fliss, I. 2003. Liposome encapsulated nisin Z: optimization, stability and reléase during milk fermentation. International Dairy Journal. 13: 325-336. • Lante, A., Crapisi, A., Pasini, G., and Scalabrini, P. 1994. Nisin released from immobilization matrices as antimicrobial agent. Bitoechnol. Letts 16: 293-298. • Lante, A., Crapisi, A., Zannoni, S., and Spettoli, P. 2000. Nisin released from membrane reactor for dairy Clostridia control. Industrie Alimentai XXXIX: 589-595. • Robinson, S. K. 1993. Regulator aspects of bacteriocin use. In Bacteriocins of Lactic Acid Bacteria. Ed. Hoover, DG and Steenson, L. R. pp 233-247. London: Academic Press. • Shahidi, F. and Han, X. G. 1993. Encapsulation of food ingredients. Critical Review in Food Science and Nutrition 33: 501-547. • Wan, J., Hickey, M. W. and Coventry, M. J. 1995. Continuous production of bacteriocins, brevicin, nisin and pediocin, using calcium alginate-immobilized bacteria. Journal of Applied Bacteriology 79: 6712-676. • Wan, J., Gordon, J. B., Muirhead, K., Hickey, M.W., and Coventry, M.J. 1997. Incorporation of nisin in micro-particles of calcium alginate. Letters in Applied Microbiology 24: 153-158. • WO 02/094224 A1. Bioactive agent + bioactive carbohydrate polymer. Not relevant • WO 9856402. Ambi. Oral formulation of nisin with a salt in a coating to give a relapse into the colon to treat bacterial infections. • WO 9720473. Wrigley. Chewing gum with improved flavor using nisin, coating to make a pellet. • GB2388581A. Microcapsules and method for preparing them. Encapsulation method.
Degradation of nisin by proteases · Alifax, R. and Chevalier, R. 1962. Study of the nisinase produced by
Streptococcus thermophilus. J. Dairy Res 29: 233 • Campbell, L. L. 1959. Effect of subtilin and nisin on spores of Bacillus coagulans. J. Bacteriol. 77: 766. • Jarvis, B. and Mahoney, R. R. 1969. Inactivation of nisin by alpha chymotrypsin. Journal of Dairy Science 52: 1448-1450. • Jarvis, B. 1967. Resistance to nisin and production of nisin-inactivating enzymes by several Bacillus species. J. Gen Microbiol 47: 33. All publications mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the spirit and scope of the invention. Although the invention has been described in connection with specific preferred embodiments, it will be understood that the invention, as claimed, should not be limited to said specific embodiments. In fact, various modifications of the modes described to carry out the invention that are obvious to those skilled in chemistry, biology, food science or related fields, are intended to be within the scope of the appended claims.
45. - The composition according to any of claims 37 to 43, further characterized in that the vehicle comprises an emulsifier. 46. - A protected food comprising (i) a food, and (ii) an antimicrobial material according to any of claims 1 to 36 or a composition according to any of claims 37 to 45. 47. - The food protected in accordance with the claim
46, further characterized in that the food is selected from raw meat, cooked meat, raw poultry products, cooked poultry products, raw seafood products, and cooked seafood products. 48. - Protected food in accordance with the claim
47, further characterized because the food is raw meat. 49. - The protected food according to claim 47 further characterized in that the food is a raw or cooked poultry product. 50. - The protected food according to claim 46 or 47 further characterized in that the food comprises whole meat muscle. 51. - A method for introducing an antimicrobial material into a food comprising (i) providing the antimicrobial material in an encapsulated form comprising a core of antimicrobial material and shell of encapsulating material, (ii) introducing antimicrobial material
Claims (2)
1 - . 1 - An antimicrobial material in an encapsulated form, comprising (i) a core comprising an antimicrobial material and (i) a shell of encapsulating material, wherein the shell of encapsulating material is impermeable to the antimicrobial material.
2 - The antimicrobial material according to claim 1, further characterized in that the antimicrobial material is an antibacterial material. 3. The antimicrobial material according to claim 1 or 2, further characterized in that the antimicrobial material is a bacteriocin. 4. - The antimicrobial material according to claim 1, further characterized in that the antimicrobial material is an antifungal material. 5. - The antimicrobial material according to claim 1, further characterized in that the antimicrobial material is at least natamycin. 6. - The antimicrobial material according to claim 3, further characterized in that the bacteriocin is selected from bacteriocins containing lanthionine, bacteriocins derived from 137 Lactococcus, bacteriocins derived from Streptococcus, bacteriocins derived from Pediococcus, bacteriocins derived from Lactobacillus, bacteriocins derived from Carnobacterium, bacteriocins derived from Leuconostoc, bacteriocins derived from Enterococcus and mixtures thereof. 7. - The antimicrobial material according to claim 1, further characterized in that the antimicrobial material is at least nisin. 8. - The antimicrobial material according to any of the preceding claims, further characterized in that the antimicrobial material is present in an amount to provide a microbicidal or microbiostatic effect. 9. - The antimicrobial material according to claim 8, further characterized in that the microbicidal or microbiostatic effect is a bactericidal or bacteriostatic effect. 10. - The antimicrobial material according to claim 9, further characterized in that the bactericidal or bacteriostatic effect is with respect to gram-positive bacteria. 11. - The antimicrobial material according to claim 9, further characterized in that the bactericidal or bacteriostatic effect is with respect to an organism selected from Bacillus species, Clostridium species, Listeria monocytogenes, lactic acid bacteria, Leuconostoc species, Carnobacterium , Enterococcus; Brochothrix 138 thermosphacta and Lactobacillus. 12. - The antimicrobial material according to claim 9, further characterized in that the bactericidal or bacteriostatic effect is with respect to Listeria monocytogenes. 13. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. 14. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material. 15. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material under predetermined conditions. 16. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is selected to release the antimicrobial material from the encapsulated antimicrobial material on contact with a food. 17. - The antimicrobial material according to claim 16, further characterized in that the food is a marinade. 18. - Antimicrobial material in accordance with any of 139 the preceding claims, further characterized in that the shell of the encapsulated antimicrobial material is capable of withstanding an injection. 19. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell of the encapsulated antimicrobial material is able to withstand a pressure greater than 1.5 bar. 20. - The antimicrobial material according to any of the preceding claims, further characterized in that the encapsulated antimicrobial material is a particle form. 21. - The antimicrobial material according to any of the preceding claims, further characterized in that the encapsulated antimicrobial material has an average particle size of less than 150 μ ??. 22. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is selected to provide sustained release of the antimicrobial material from the encapsulated antimicrobial material. 23. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material. 24. - The antimicrobial material according to claim 18 further characterized in that the degeneration is by one or 140 plus selected factors of thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation. 25. - The antimicrobial material according to any of the preceding claims, further characterized in that the shell is or comprises material selected from fats, emulsifiers, waxes (of animal, vegetable, mineral or synthetic origin), lipid-forming liposomes, hydrocolloids , natural or synthetic polymers and mixtures thereof. 26. - The antimicrobial material according to claim 25, further characterized in that the lipid is a glycerophospholipid and / or esteral. 27. - The antimicrobial material according to claim 25 or 26, further characterized in that the fat is a triglyceride. 28. - The antimicrobial material according to claim 27, further characterized in that the triglyceride is a vegetable triglyceride. 29. - The antimicrobial material according to any of claims 25 to 28, further characterized in that the emulsifier is selected from polysorbates, monoglycerides, diglycerides, acetic acid esters of mono- and diglycerides, esters of tartaric acid of mono- and diglycerides and citric acid esters of mono- and diglycerides. 30. - The antimicrobial material according to any of claims 25 to 29, further characterized in that the hydrocolloid is 141 interlaced 31. - The antimicrobial material according to claim 30 further characterized in that the hydrocolloid is carrageenan. 32. - The antimicrobial material according to any of the preceding claims, further characterized in that the encapsulated antimicrobial material is prepared or obtainable by a process selected from spray cooling, and fluidized bed coating. 33. - The antimicrobial material according to any of the preceding claims, further characterized in that the encapsulated antimicrobial material further comprises a chelator. 34. - The antimicrobial material according to claim 33, further characterized in that the chelator is selected from EDTA, citric acid, monophosphates, diphosphates, triphosphates and polyphosphates. 35. - The antimicrobial material according to claim 33 or 34, further characterized in that the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the antimicrobial material. 36. - The antimicrobial material according to claim 33, 34 or 35, further characterized in that the chelator improves the antimicrobial activity and / or antimicrobial spectrum of the antimicrobial material with respect to gram-negative bacteria. 142 37. - A . composition comprising (i) an antimicrobial material according to any of the preceding claims (i) a vehicle. 38. - The composition according to claim 37, further characterized in that the vehicle is or comprises brine. 39. The composition according to claim 37 or 38, further characterized in that the vehicle and the encapsulated antimicrobial material have substantially the same density. 40. The composition according to claim 37, further characterized in that the encapsulated antimicrobial material is modified to have substantially the same density as the vehicle. 41. - The composition according to claim 40, further characterized in that the encapsulated antimicrobial material is modified by contacting the encapsulated antimicrobial material with oil. 42. - The composition according to claim 41, further characterized in that the oil is brominated oil. 43. - The composition according to claim 41 or 42, further characterized in that the vehicle is modified to have substantially the same density as the encapsulated antimicrobial material. 44. - The composition according to claim 43 further characterized in that the vehicle comprises xanthan gum. encapsulated inside or on the food. 52. - The method according to claim 51, further characterized in that the encapsulated antimicrobial material is introduced into or onto the food by (a) injecting the antimicrobial material encapsulated in the food or (b) stirring the encapsulated antimicrobial material with the food . 53. - The method according to claim 51 or 52, further characterized in that the encapsulated antimicrobial material is introduced into the food by injecting the antimicrobial material encapsulated in the food. 54. - The method according to claim 51 or 52, further characterized in that the encapsulated antimicrobial material is introduced into or onto the food when the antimicrobial material encapsulated with the food is returned. 55. - The method according to claim 46, further characterized in that (i) the antimicrobial material is at least nisin, (i) the antimicrobial material is present in an amount to provide a microbicidal or microbiostatic effect with respect to Listeria monocytogenes, (iii) the shell is selected to prevent, reduce or inhibit degeneration or inactivation of the antimicrobial material by one or more factors selected from thermal degradation, pH-induced degradation, protease degradation and glutathione adduct formation; and (iv) the food is selected from raw meat products, cooked meat products, raw seafood products, cooked seafood products, raw poultry products and cooked poultry products. 56. - A food prepared by a process as defined in any of claims 46 to 55. 57. - A food obtainable by a process as defined in any of claims 46 to 55.
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GB0319817A GB2388581A (en) | 2003-08-22 | 2003-08-22 | Coated aqueous beads |
GB0323335A GB0323335D0 (en) | 2003-10-06 | 2003-10-06 | Composition |
US53305303P | 2003-12-30 | 2003-12-30 | |
US56027004P | 2004-04-08 | 2004-04-08 | |
US10/820,147 US20050042341A1 (en) | 2003-08-22 | 2004-04-08 | Natamycin dosage form, method for preparing same and use thereof |
PCT/GB2004/003397 WO2005018322A1 (en) | 2003-08-22 | 2004-08-06 | Encapsulated antimicrobial material |
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MXPA06002060A MXPA06002060A (en) | 2003-08-22 | 2004-08-06 | Composition comprising a bacteriocin and an extract from a plant of the labiatae family. |
MXPA06002062A MXPA06002062A (en) | 2003-08-22 | 2004-08-06 | Microcapsules. |
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MXPA06002062A MXPA06002062A (en) | 2003-08-22 | 2004-08-06 | Microcapsules. |
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US9883689B2 (en) * | 2015-04-17 | 2018-02-06 | Kerry Luxembourg S.à.r.l. | Composition and methods to control the outgrowth of pathogens and spoilage microorganisms in high moisture and low sodium systems |
US11523626B2 (en) * | 2018-04-13 | 2022-12-13 | Pontificia Universidad Catolica De Chile | Synergic composition for keeping fish and seafood fresh |
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AU631803B2 (en) * | 1988-06-22 | 1992-12-10 | Applied Microbiology, Inc | Nisin compositions for use as enhanced, broad range bactericides |
NZ244737A (en) * | 1989-02-21 | 1993-09-27 | Viskase Corp | Food packaging polymeric film containing antibiotic material; method of treating foodstuff and food casing therefor |
US4980163A (en) * | 1989-03-01 | 1990-12-25 | Public Health Research Institute Of The City Of New York | Novel bacteriocin compositions for use as enhanced broad range bactericides and methods of preventing and treating microbial infection |
US6126974A (en) * | 1998-03-13 | 2000-10-03 | Protein X Technologies Int'l Inc. | Anti-caking anti-mycotic food ingredient and process for producing the same |
WO2001005254A2 (en) * | 1999-07-14 | 2001-01-25 | Rhodia Inc. | An antibacterial composition for control of gram positive bacteria in food |
US6291436B1 (en) * | 1999-07-16 | 2001-09-18 | International Fiber Corporation | Direct action anti-mycotic |
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