Inhibiting fungal growth by manganese consumption

Abstract

The present invention provides a method for controlling the growth of harmful microorganisms by restricting their access to manganese. More specifically, the present invention provides a method for inhibiting or delaying the growth of yeasts and molds by reducing the manganese concentration in a product, preferably in a food product. The present invention also provides a manganese scavenger and its use for inhibiting or delaying fungal growth. The manganese scavenger can be bacterial cells, specifically lactic acid bacteria cells, and more specifically, Lactobacillus rhamnosus alone or in combination with Lactobacillus paracasei.

Description

Field of the Invention

[0001] The present invention belongs to the field of microbiology and relates to methods for controlling fungal spoilage. The present invention also relates to foods and preparations. Background Art

[0002] A major problem in the food industry is spoilage caused by harmful microorganisms. According to the Food and Agriculture Organization (FAO), ultimately one quarter of the calories available for human consumption are not consumed by humans. During times of food shortage, with over 800 million people suffering from hunger, the topic of food waste has become a top priority for global policymakers and food manufacturers. In addition to the negative social and economic impacts on society, wasted food also has many associated environmental impacts, including unnecessary greenhouse gas emissions and inefficient use of scarce resources such as water and land.

[0003] Yeasts and molds are very effective in causing food spoilage, which is a problem for most food manufacturers. Spoilage caused by yeasts and molds is clearly visible as mold spots or discoloration on the surface of foods, allowing it to be addressed before consumption. Yeasts tend to grow in a planktonic form in food and beverage matrices and they tend to ferment sugars and grow well under anaerobic conditions. In contrast, molds tend to grow on the surface of products in the form of visible mycelia composed of cells.

[0004] Especially in the dairy industry, 29 million tons of dairy products are wasted in Europe every year. One of the main challenges in keeping dairy products fresh is controlling yeast and mold contamination, which are naturally present everywhere, especially when the cold chain is interrupted from production to the consumer's table.

[0005] Due to economic and environmental reasons, there has been a continuing need for novel or improved methods for effectively controlling yeast and mold contamination. Summary of the Invention

[0006] The inventors of the present invention sought to find effective methods for controlling fungal contamination and surprisingly identified manganese as an important growth limiting condition for their growth. The present invention provides a new method for inhibiting fungal growth by restricting the free manganese available to yeasts or molds. The present invention is in part based on the surprising finding that by reducing the content of free manganese in foods, for example by using a manganese scavenger to remove free manganese, contamination by yeasts and / or molds can be reduced or delayed. In addition, the inventors have also demonstrated that common yeasts and molds are sensitive to the methods described herein.

[0007] Manganese is considered essential for human health and is thus an essential trace element. Manganese is crucial for the normal functions of humans and animals, as it is required for the action of many cellular enzymes such as manganese superoxide dismutase, pyruvate carboxylase, and it can activate many other enzymes such as kinases, decarboxylases, transferases, and hydrolases.

[0008] Manganese occurs naturally in many food sources, including leafy vegetables, nuts, cereals, and animal products. The typical range of manganese concentration in common foods is, for example, 0.4 - 40 ppm in cereal products, 0.1 - 4 ppm in meat, poultry, fish, and eggs, and 0.4 - 7 ppm in vegetable products.

[0009] In addition to being used as a dietary supplement, manganese is sometimes added to fermented products as an active ingredient to enhance the growth of Bifidobacterium in milk (see, for example, WO2017 / 021754, Compagnie Gervais Danone, France). However, the present inventors have found that this may have an adverse effect, and it may be advantageous to limit the concentration of manganese in foods in order to prevent or delay the growth of harmful microorganisms.

[0010] To solve the problem of microbial spoilage, in a first aspect, the present invention provides a method for inhibiting or delaying the growth of one or more fungi in a product, the method comprising the step of reducing the free manganese present in the product. The method of the present invention can reduce the free manganese concentration. In a preferred embodiment, one or more manganese scavengers are added to reduce the free manganese. The free manganese concentration is preferably reduced to less than about 0.01 ppm, for example less than about 0.008 ppm, or less than about 0.003 ppm. Using this method, a product can be obtained in which harmful yeast and / or mold can hardly grow. The product is characterized by a free manganese concentration of less than about 0.01 ppm, for example 0.009 ppm, 0.008 ppm, 0.007 ppm, 0.006 ppm, 0.005 ppm or lower. The method further comprises the steps of measuring the free manganese concentration in the product and obtaining a value of less than about 0.01 ppm.

[0011] In particular, the present invention provides a method for inhibiting or delaying the growth of yeast and / or mold in fermented foods prepared from milk, such as yogurt or cheese. The method is characterized by the step of reducing the manganese concentration in the food to deprive the yeast and / or mold of manganese, thereby delaying or inhibiting their growth in the food.

[0012] In a preferred embodiment, the present invention provides a method for inhibiting or delaying the growth of Torulaspora spp, Cryptococcus spp, and Rhodotorula spp in a product, comprising the step of reducing the free manganese present in the product.

[0013] In a second aspect, the present invention provides a method for preparing a product such as a food product, the method comprising reducing the free manganese present in the product. The free manganese concentration can be reduced by the method described herein or by other methods known to those skilled in the art. In a preferred embodiment, one or more manganese scavengers are added to reduce the free manganese. The free manganese concentration is preferably reduced to less than about 0.01 ppm, such as less than about 0.005 ppm, or less than about 0.003 ppm. Using this method, a product with a free manganese concentration of less than about 0.01 ppm can be obtained.

[0014] In a third aspect, the present invention provides a product obtained by the method described herein, such as a food product. In one embodiment, the present invention provides a method for providing a product, comprising the steps of reducing the free manganese in the product and obtaining the product, wherein the free manganese concentration in the product is less than about 0.01 ppm.

[0015] In another aspect, the present invention provides the use of one or more manganese scavengers in inhibiting or delaying fungal growth and in the production of food products. The manganese scavenger has the effect of reducing the free manganese available to yeasts and molds in the product, thereby inhibiting or delaying their growth.

[0016] In another aspect, the present invention provides manganese scavengers, the selection of manganese scavengers, and their use in manganese uptake. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 and Figure 2 shows the growth of two different strains of Debaryomyces hansenii in a chemically defined medium with different manganese concentrations at pH 6.5 (open circles) or 4.5 (black squares). Growth was measured after incubation at 17 °C for 6 days and determined by absorbance at 600 nm.

[0018] Figure 3 and Figure 4 shows the growth of two different strains of Debaryomyces hansenii in the aqueous phase of fermented milk in the presence of a manganese scavenger and with different manganese concentrations added. After incubation at 17 °C for 7 days, the absorbance at 600 nm was measured. Compared to the reference without added manganese (squares), the growth of Debaryomyces hansenii (strain 1) after adding manganese to the aqueous phase is as Figure 3As shown, the growth of Debaryomyces hansenii (strain 2) is as Figure 4 shown (open circles). The mean and standard deviation of technical replicates are shown for n = 6 (A) and n = 3 (B).

[0019] Figure 5 The growth of Debaryomyces hansenii (strain 2) in fermented milk is shown after addition of different manganese concentrations from 6 ppm (top row) to 0.000006 ppm (bottom row), and the fermented milk was prepared with and without different scavengers.

[0020] Figure 6 The growth of three different yeasts on plates prepared from milk is shown, and the milk was fermented with a single starter culture (reference, top row) or with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were added. Three target contaminants were added at three different concentrations (Column A: Debaryomyces hansenii (strain 2), Column B: Cryptococcus fragicola, Column C: Debaryomyces hansenii (strain 1)): 1x10 3 cfu / spot (top row of the plate), 1x10 2 cfu / spot (middle row of the plate) and 1x10 1 cfu / spot (bottom row of the plate).

[0021] Figure 7 The growth of three different molds on plates prepared from milk is shown, and the milk was fermented with a single starter culture (reference, top row) or with a manganese scavenger (bottom row). As shown in the figure, different manganese concentrations were added. As shown above the picture, different manganese concentrations were added. Three target contaminants were added at a concentration of 500 spores / spot (A: Penicillium brevicompactum, B: Penicillium crustosum, and C: Penicillium solitum). The plates were incubated at 7 ± 1 °C for 25 days.

[0022] Figure 8 The growth of three different molds on plates prepared from milk is shown, and the milk was fermented with a single starter culture (reference, top row) or with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were added. Three target contaminants were added at a concentration of 500 spores / spot (A: Penicillium brevicompactum, B: Penicillium crustosum, and C: Penicillium solitum). The plates were incubated at 22 ± 1 °C for 8 days.

[0023] Figure 9Shows the growth of three different molds on plates prepared from milk, which was fermented by a starter culture alone (reference, top row), or alternatively by a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were further added. Three target contaminants were added at a concentration of 500 spores / spot (A: Penicillium carneum, B: Penicillium paneum, and C: Penicillium roqueforti). The plates were incubated at 7 ± 1 °C for 25 days.

[0024] Figure 10 Shows the growth of three different molds on plates prepared from milk, which was fermented by a starter culture alone (reference, top row), or alternatively by a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were further added. Three target contaminants were added at a concentration of 500 spores / spot (A: Penicillium carneum, B: Penicillium paneum, and C: Penicillium roqueforti). The plates were incubated at 22 ± 1 °C for 8 days.

[0025] Figure 11 Shows an exemplary phylogenetic tree of the MntH manganese transporter of selected species of the genus Lactobacillus.

[0026] Figure 12 Shows the growth of Hanseniaspora uvarum strains (strain 1 or strain 2) in the aqueous phase, which was filtered through a 1.2 μm filter (filtered water), preferably the aqueous phase of fermented milk, with different EDTA concentrations, with and without other manganese scavengers, preferably manganese scavenger 1, and with and without 6 ppm of manganese. The absorbance at 600 nm was measured after incubation at 17 °C for 7 days compared to the reference. Samples were prepared in triplicate.

[0027] Figure 13 Shows the growth of Hanseniaspora uvarum strains (strain 1 or strain 2) in the aqueous phase, which was filtered through a 0.2 μm filter (sterile water), preferably the aqueous phase of fermented milk, with different EDTA concentrations, with or without other manganese scavengers, preferably manganese scavenger 1, and with and without 0.6 ppm of manganese. The absorbance at 600 nm was measured after incubation at 17 °C for 7 days compared to the reference. Samples were prepared in triplicate.

[0028] Figure 14Shows the growth of Hansenula polymorpha strains (strain 1 or strain 2) in yogurt supplemented with 6 ppm manganese, said yogurt having and not having different manganese scavengers, such as bacteria, preferably lactic acid bacteria, and / or chemical chelating materials, such as EDTA. The concentration of EDTA is 14 mg / ml (top row), 7.10 mg / ml, 3.55 mg / ml, 1.78 mg / ml, 0.89 mg / ml, 0.44 mg / ml, 0.22 mg / ml to 0 mg / ml (bottom row). Detailed Description

[0029] Food loss is a major problem worldwide - approximately one-third of all food produced for human consumption is lost or wasted. The causes of significant food losses globally are diverse, but microbial spoilage, which affects the sensory quality of products (appearance, texture, taste, and aroma), plays an important role. Since fungi can grow in diverse and even harsh environments, they are the main spoilage microorganisms found at all stages of the food processing chain. Therefore, it is crucial to reduce food loss by controlling fungal contamination.

[0030] In response to this need, the present invention provides a new method for inhibiting or delaying fungal growth in products. The method is based on the surprising discovery that low free manganese concentrations can act as a limiting factor for yeast and / or mold growth. Trace amounts of manganese are present in nature and in many of our consumer products. However, it has not been reported that manipulating the free manganese concentration can effectively control microbial spoilage. Based on this unexpected discovery, it is foreseeable that this spoilage prevention strategy is applicable even to products other than food and extends to other products that are commonly vulnerable to microbial contamination, such as feed products, biological products, health products, pharmaceutical products, etc.

[0031] In a first aspect, the present invention provides a method for inhibiting or delaying fungal growth in a product, comprising consuming the free manganese in the product to a concentration below about 0.01 ppm.

[0032] Generally, inhibition refers to a partial or complete reduction in the function and activity of cells or microorganisms. As used herein, the terms "to inhibit" and "inhibiting" with respect to yeast and molds refer to the growth, number, or concentration of yeast and molds being the same or reduced. This can be measured by any method known in the microbiology field. Inhibition can be observed by comparing the fungal growth, number, or concentration in or on a product with reduced free manganese to a control. The control can be the same product, but with no reduction in free manganese.

[0033] The term "delay" generally refers to stopping, postponing, hindering, or causing something to occur more slowly than normal. As used herein, "delaying fungal growth" refers to an action that postpones fungal growth. This can be observed by comparing the time it takes for fungal growth to reach a given level in two products, one of which has reduced manganese while the other has not (but is otherwise the same).

[0034] In some embodiments, "delaying fungal growth" means a delay of 7 days, such as 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60 days.

[0035] Fungi are members of the kingdom Fungi. Various methods known to those skilled in the art can be used to measure the growth of fungi. For example, the growth of fungi can be measured by the density or size of colonies, the number of cells, changes in mycelial mass, spore production, hyphal growth, colony-forming units (CFU), etc., depending on the type of fungus and the product to which the application method is applied. The growth of fungi can also be observed by measuring changes in the concentration of nutrients or metabolites, such as carbon dioxide release and oxygen uptake. The terms "inhibition of fungal growth" or "inhibiting growth of fungi" refer to inhibiting the proliferation of fungal cells. The terms "delay of fungal growth" or "delaying growth of fungi" refer to slowing down the proliferation of fungal cells. For example, this can be observed by measuring the growth of fungi and comparing it to a control. Such a control can be, for example, a product to which no manganese scavenger has been applied. Methods for determining fungal growth inhibition or delay are known to those skilled in the art.

[0036] According to the present invention, "free manganese" or sometimes just "manganese" refers to manganese that is present in a product (i.e., forms part of the product, such as inside the product or on the surface of the product) and can be utilized or absorbed by fungi, including yeasts and molds. For example, free manganese refers to the manganese present in the food matrix of a product.

[0037] In a preferred embodiment, the present invention relates to a method for inhibiting or delaying fungal growth in food, comprising reducing the concentration of free manganese in the food matrix of the food. As used herein, the term "food matrix" refers to the composition and structure of food. It is based on the concept that nutrients are contained in a continuous medium.

[0038] The term "reduce" or "reducing" generally refers to a decrease in the amount of a substance in a given context. As used herein, the term "to reduce free manganese" or "reducing free manganese" refers to a decrease in the amount of manganese present in a product that can be taken up by fungi, including yeasts and molds.

[0039] For example, this can be done by removing manganese present in the product or in the materials that make up the product. For example, this can be done by subjecting the raw materials to ion exchange chromatography to remove manganese, thereby reducing the concentration in the final product.

[0040] Once inside, the fungi rapidly colonize, increase in population, and take up nutrients from their surrounding environment. In some embodiments, it is within the spirit of the invention that the reducing step be carried out on a portion of the product, for example on the outside of the product, such as a coating or outer layer, assuming that the fungi can first come into contact with the product on the surface. In such a case, the reducing step still results in an overall decrease in the concentration in the product.

[0041] The manganese concentration or manganese level as used herein is expressed as parts per million ("ppm") based on weight / weight. Reducing the free manganese in a product to a concentration below a certain value means reducing the free manganese in the product or a portion thereof, thereby reducing the concentration of free manganese by weight in the entire product. Methods for determining trace elements such as manganese are known in the art and are described, for example, in Nielsen, S. Suzanne, ed. Food analysis. Vol. 86. Gaithersburg, MD: Aspen Publishers, 1998.

[0042] When applying the method of the present invention, those skilled in the art can first determine the manganese content present in the product to be processed. The concentration of manganese in food has been well studied and can be found in national food composition databases, such as the Danish Food Composition Databank and the Canadian Nutrient Files. Generally, manganese in milk is present at a concentration of at least 0.03 ppm, making dairy products vulnerable to fungal contamination. The manganese content in cow's milk has been reported to be 0.04 - 0.1 ppm, and the manganese content in goat or sheep milk has been reported to be as high as 0.18 ppm (Muehlhoff et al., Milk and dairy products in human nutrition. Food and Agriculture Organization of the United Nations (FAO), 2013). As for fermented dairy products such as cheese, due to the concentration process of milk, the manganese content usually increases, often up to 10 times or more. Different contents of various cheese types have been reported, for example, ricotta cheese is about 0.06 ppm, cream cheese is 0.11 ppm, brie is 0.34 ppm, mozzarella is 0.3 ppm, cottage cheese is 0.7 ppm, gouda is 0.68 ppm, and cheddar cheese is 0.74 ppm (Smit, L.E., et al. The nutritional content of South African cheeses. ARC - Animal Improvement Institute, 1998; Gebhardt, Susan, et al. "USDA national nutrient database for standard reference, release 12." United States Department of Agriculture, Agricultural Research Service, 1998).

[0043] Preferably, the free manganese in the product is reduced to a concentration of less than about 0.01 ppm, such as less than about 0.009 ppm, less than about 0.008 ppm, less than about 0.007 ppm, less than about 0.006 ppm, less than about 0.005 ppm, less than about 0.004 ppm, less than about 0.003 ppm, less than about 0.002 ppm, less than about 0.001 ppm, less than about 0.0009 ppm, less than about 0.0008 ppm, less than about 0.0007 ppm, less than about 0.0006 ppm, less than about 0.0005 ppm, less than about 0.0004 ppm, less than about 0.0003 ppm, or lower.

[0044] As used herein, the term "about" means a value slightly beyond the cited value, i.e., plus or minus 0.1% to 10%. Accordingly, the scope of the present invention also includes concentrations slightly beyond the stated ranges.

[0045] In one embodiment, the present invention provides a method for inhibiting or delaying fungal growth in a product, preferably a food product, comprising the steps of:

[0046] - reducing the free manganese in the product, and

[0047] - obtaining a product in which the free manganese concentration is less than about 0.01 ppm.

[0048] The method further comprises the step of measuring the free manganese concentration. This can be done after the reducing step to determine whether the concentration of free manganese has been reduced. In one embodiment, the present invention provides a method for inhibiting or delaying fungal growth in a food product, comprising reducing the free manganese in the food product to a concentration of less than about 0.01 ppm in the food product, measuring the free manganese in the food product, and optionally obtaining a value less than 0.01 ppm.

[0049] In one embodiment, the present invention provides a method for inhibiting or delaying fungal growth in a product, comprising the steps of:

[0050] - reducing the free manganese in the product to a concentration of less than about 0.01 ppm in the product,

[0051] - measuring the concentration of free manganese in the product and obtaining a value less than 0.01 ppm.

[0052] Methods for measuring low concentrations of manganese are well known to those skilled in the art. Such methods include atomic absorption spectrometry, atomic emission spectrometry, mass spectrometry, neutron activation analysis, and X-ray fluorescence spectrometry (see, for example, Williams et al. "Toxicological profile for manganese." (2012)).

[0053] Preferably, the manganese concentration is measured according to the standard procedures described below: "Foodstuffs - Determination of trace elements - Pressure digestion" in European Standard EN 13805:2014 published by the European Committee for Standardization, or "Water quality - Determination of selected elements by inductively coupled plasma optical emission spectrometry (ICP - OES)" in ISO 11885:2007 published by the International Organization for Standardization.

[0054] Fungus

[0055] The inventors of the present invention surprisingly found that both yeast and mold can be inhibited by manganese consumption. In a preferred embodiment, the present invention provides a method for inhibiting or delaying the growth of yeast in a product, preferably a food, comprising the step of reducing the free manganese in the product. In another preferred embodiment, the present invention provides a method for inhibiting or delaying the growth of mold in a product, preferably a food, comprising the step of reducing the free manganese in the product.

[0056] In one embodiment, the method is used to inhibit the growth of yeasts, such as species of Candida spp., Meyerozyma spp., Kluyveromyces spp., Pichia spp., Galactomyces spp., Trichosporon spp., Sporidiobolus spp., species of Torulaspora, Cryptococcus spp., Saccharomyces spp., Yarrowia spp., Debaryomyces spp., and Rhodotorula spp. Preferably, the fungus is a yeast selected from the group consisting of species of Torulaspora, Cryptococcus, Saccharomyces, Yarrowia, Debaryomyces, Candida, and Rhodotorula. More preferably, the fungus is a yeast selected from the group consisting of Torulaspora delbrueckii, Cryptococcus laurentii, Saccharomyces cerevisiae, Yarrowia lipolytica, Debaryomyces hansenii, and Rhodotorula mucilaginosa.

[0057] In one embodiment, the method is used to inhibit the growth of molds. Preferably, the fungus is a mold selected from the group consisting of species of Aspergillus spp., Cladosporium spp., Didymella spp., or Penicillium spp. More preferably, the fungus is a mold selected from the group consisting of Penicillium brevicompactum, Penicillium crustosum, Penicillium divaricatum, Penicillium carneum, Penicillium vinaceum, and Penicillium roqueforti.

[0058] Manganese removal

[0059] Methods for removing manganese are known in the art. Manganese is a common contaminant in many mine waters, groundwaters, and freshwaters. In wastewater treatment, manganese ions can be chemically removed from wastewater by oxidation to MnO2, adsorption, or precipitation as carbonates.

[0060] As an alternative option, manganese removal can involve biological processes as an alternative to chemical pathways. The role of microbial activity in the remediation of manganese-contaminated water has been described in various literature, such as Burger et al. Manganese removal during bench-scale biofiltration. Water Research. 2008; 42(19):4733–4742; Johnson et al. Rapid manganese removal from mine waters using an aerated packed-bed bioreactor. Journal of Environmental Quality. 2005; 34(3):987–993; Tekerlekopoulou et al. "Removal of ammonium, iron and manganese from potable water in biofiltration units: a review." Journal of Chemical Technology and Biotechnology 88.5(2013):751-773; Patil et al. “A review of technologies for manganese removal from wastewaters." Journal of Environmental Chemical Engineering 4.1(2016):468-487. In one embodiment, the step of reducing free manganese in the product involves using ion exchange chromatography. This is particularly applicable if the product is liquid or substantially liquid.

[0061] In a preferred embodiment, the step of reducing free manganese in the product is carried out by adding a manganese scavenging agent. As used herein, the term "manganese scavenging agent" or "manganese scavenger" refers to a material that can render manganese unavailable to yeast or mold. The material can be a chemical material, such as a chemical chelating material selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid (EGTA), diaminocyclohexanetetraacetic acid (DCTA), nitrilotriacetic acid (NTA), 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), or diethylenetriaminepentaacetic acid (DTPA). Preferably, the chemical chelating material is ethylenediaminetetraacetic acid (EDTA). The material can also be a biological material, such as bacteria.

[0062] In some preferred embodiments, the manganese scavenging agent is one or more bacterial strains. In this case, it should be noted that when measuring free manganese, this free manganese does not include the manganese found inside the cells. Instead, free manganese refers to the manganese found extracellularly, i.e., in the cell-free portion of the product, as they can be taken up by fungi. Therefore, in this case, only the extracellular manganese should be considered to measure the concentration of free manganese. For example, the concentration of free manganese can be measured by centrifuging to remove the cells (e.g., the starter culture) and obtaining the cell-free supernatant, and then measuring the manganese in the cell-free supernatant. As used herein, the term "bacterial strain" has its general meaning in the field of microbiology and refers to a genetic variant of bacteria.

[0063] In one embodiment, the present invention provides a method for inhibiting or delaying fungal growth in a product, comprising the steps of:

[0064] selecting one or more bacterial strains as manganese scavenging agents, and

[0065] - reducing the free manganese in the product to a concentration of less than about 0.03 ppm in the product by adding the manganese scavenging agent.

[0066] According to a preferred embodiment of the present invention, the method comprises selecting a bacterial strain having manganese uptake activity as the manganese scavenging agent. The selection is based on whether the bacterial strain has a manganese transport system.

[0067] Manganese is involved in many key biological processes and is ubiquitous in all living organisms. Manganese also helps prevent oxidative stress and can contribute to the catalytic detoxification of reactive oxygen species. Many bacteria have developed elaborate acquisition systems to scavenge essential metals from the environment using low- and high-affinity transport systems for chelated or free metals. The manganese taken up by bacteria forms large complexes of indialyzable polyphosphorylated protein aggregates in proteins, which can reach very high intracellular concentrations.

[0068] The manganese transport systems have been studied, for example as described in Kehres et al., "Emerging themes in manganese transport, biochemistry and pathogenesis in bacteria." FEMS microbiology reviews 27.2-3 (2003): 263-290.

[0069] In one embodiment, a bacterial strain having manganese uptake activity comprises a bacterial Mn2+ transporter. The Mn2+ transporter can be an ABC transporter (such as SitABCD and YfeABCD), or a proton-dependent Nramp-related transport system, which belongs to the families designated as TC#3.A.1.15 and TC#2.A.55 in the transporter classification system given in the Transporter Classification Database (M. Saier; U of CA, San Diego, Saier MH, Reddy VS, Tamang DG, Vastermark A. (2014)). The TC system is a classification system for transporters, similar to the enzyme classification system of the Enzyme Commission (EC). The Transporter Classification (TC) system is a terminology system for transporter classification approved by the International Union of Biochemistry and Molecular Biology. TCDB is accessible for free at http: / / www.tcdb.org and provides several different ways of accessing data, including stepwise access to the hierarchical classification, direct search by sequence or TC number, and full text search.

[0070] In one embodiment, the method comprises selecting a bacterial strain as a manganese scavenger, the bacterial strain comprising a protein belonging to the family designated as TC#3.A.1.15 (the manganese chelate uptake transporter (MZT) family).

[0071] For example, a manganese scavenger is a bacterial strain that contains a manganese chelate uptake transporter called TC#3.A.1.15.2, TC#3.A.1.15.6, TC#3.A.1.15.8, TC#3.A.1.15.14 or a functional variant thereof.

[0072] Although ABC transporters are mainly active at higher pH values, proton-driven transporters may be more active under acidic conditions. This makes them particularly useful as manganese scavengers in fermented foods. Thus, in one embodiment, a bacterial strain is selected that contains a protein belonging to the family called TC#2.A.55 (the metal ion (Mn 2+ -iron) transporter (Nramp) family).

[0073] The step of selecting one or more bacterial strains as manganese scavengers includes determining whether the one or more bacterial strains contain a manganese transporter called TC#2.A.55 or a functional variant thereof.

[0074] More preferably, the transporter used as a manganese scavenger belongs to the subfamily called TC#2.A.55.2 or the subfamily called TC#2.A.55.3.

[0075] For example, a manganese scavenger is a bacterial strain that contains a metal ion (Mn 2+ -iron) transporter (Nramp) called TC#2.A.55.3.1, TC#2.A.55.3.2, TC#2.A.55.3.2, TC#2.A.55.3.3, TC#2.A.55.3.4, TC#2.A.55.3.5, TC#2.A.55.3.6, TC#2.A.55.3.7, TC#2.A.55.3.8 or TC#2.A.55.3.9 or a functional variant thereof, as a manganese scavenger.

[0076] Most preferably, the method includes selecting a bacterial strain that contains a protein called TC#2.A.55.2.6 or a functional variant thereof as a manganese scavenger.

[0077] Preferably, the manganese scavenger is selected from the group consisting of Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus sakei, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus alimentarius, Pediococcus acidilactici, Lactobacillus rhamnosus, and Lactobacillus kefiri.

[0078] The term "functional variant" refers to a protein variant that has substantially similar biological activity, namely manganese uptake activity.

[0079] As used herein, "variant" refers to a variant form of a protein that shares at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with a specific nucleic acid or amino acid sequence of the protein.

[0080] The present invention further provides the polypeptide sequence of the manganese transporter for selecting a suitable manganese scavenger to implement the present invention.

[0081] In a preferred embodiment, the manganese scavenger is a bacterial strain comprising a polypeptide or a functional variant thereof, the polypeptide having the sequence of SEQ ID NO:1 (MASEDKKSKREHIIHFEDTPSKSLDEVNGSVEVPHNAGFWKTLAAYTGPGILVAVGYMDPGNWITSIAGGASFKYSLLSVILISSLIAMLLQAMAARLGIVTGRDLAQMTRDHTSKAMGGFLWVITELAIMATDIAEIIGSAIALKLLFNMPLIVGIIITTADVLILLLLMRLGFRKIEAVVATLVLVILLVFAYEVILAQPNVPELLKGYLPHADIVTNKSMLYLSLGIVGATVMPHDLFLGSSISQTRKIDRTKHEEVKKAIKFSTIDSNLQLTMAFIVNSLLLILGAALFFGTSSSVGRFVDLFNALSNSQIVGAIASPMLSMLFAVALLASGQSSTITGTLAGQIIMEGFIHLKMPLWAQRLLTRLMSVTPVLIFAIYYHGNEAKIENLLTFSQVFLSIALPFAVIPLVLYTSDKKIMGEFANRAWVKWTAWFISGVLIILNLYLIAQTLGFVK).

[0082] In other preferred embodiments, the manganese scavenger is a bacterial strain comprising a polypeptide having at least 55%, such as at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of SEQ ID NO:1.

[0083] Table 1 shows exemplary sequences encoding functional variants of SEQ ID NO:1, and their sequence identity to the sequence of SEQ ID NO:1.

[0084] Table 1

[0085]

[0086]

[0087]

[0088] In a preferred embodiment, the manganese scavenger is a bacterial strain comprising a polypeptide or a functional variant thereof, the polypeptide having the sequence of SEQ ID NO:2 (MARPDERLTVQREKRSLDDINRSVQVPSVYESSFFQKFLAYSGPGALVAVGYMDPGNWLTALEGGSRYHYALLSVLLMSILVAMFMQTLAIKLGVVARLDLAQAIAAFIPNWSRICLWLINEAAMMATDMTGVVGTAIALKLLFGLPLMWGMLLTIADVLVVLLFLRFGIRRIELIVLVSILTVGIIFGIEVARADPSIGGIAGGFVPHTDILTNHGMLLLSLGIMGATIMPHNIYLHSSLAQSRKYDEHIPAQVTEALRFGKWDSNVHLVAAFLINALLLILGAALFYGVGGHVTAFQGAYNGLKNPMIVGGLASPLMSTLFAFALLITGLISSIASTLAGQIVMEGYLNIRMPLWERRLLTRLVTLIPIMVIGFMIGFSEHNFEQVIVYAQVSLSIALPFTLFPLVALTNRRDLMGIHVNSQLVRWVGYFLTGVITVLNIQLAISVFV).

[0089] In other preferred embodiments, the manganese scavenger is a bacterial strain comprising a polypeptide having at least 55%, such as 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of SEQ ID NO:2.

[0090] Table 2 shows exemplary sequences encoding functional variants of SEQ ID NO:2, and their sequence identity to the sequence of SEQ ID NO:2.

[0091] Table 2

[0092]

[0093]

[0094] In a preferred embodiment, the manganese scavenger is a bacterial strain comprising a polypeptide or a functional variant thereof, said polypeptide having the sequence of SEQ ID NO:3 (MSDDHKKRHPIKLIQYANGPSLEEINGTVEVPHGKGFWRTLFAYSGPGALVAVGYMDPGNWSTSITGGQNFQYLLISVILMSSLIAMLLQYMAAKLGIVSQMDLAQAIRARTSKKLGIVLWILTELAIMATDIAEVIGAAIALYLLFHIPLVIAVLVTVLDVLVLLLLTKIGFRKIEAIVVALILVILLVFVYQVALSDPNMGALLKGFIPTGETFASSPSINGMSPIQGALGIIGATVMPHNLYLHSAISQTRKIDYKNPDDVAQAVKFSAWDSNIQLSFAFVVNCLLLVMGVAVFKSGAVKDPSFFGLFQALSDSSTLSNGVLIAVAKSGILSILFAVALLASGQNSTITGTLTGQVIMEGFVHMKMPLWARRLVTRIISVIPVIVCVMLTARDTPIQQHEALNTLMNNSQVFLAFALPFSMLPLLMFTNSKVEMGDRFKNTGWVKVLGWISVLGLTGLNLKGLPDSIAGFFGDHPTATQTNMANIIAIVLIVAILALLAWTIWDLYKGNQRYEAHLAAVADEKEAKADVDEQ).

[0095] In other preferred embodiments, the manganese scavenger is a bacterial strain comprising a polypeptide having at least 55%, such as at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of SEQ ID NO:3.

[0096] Table 3 shows exemplary sequences encoding functional variants of SEQ ID NO:3, and their sequence identities to the sequence of SEQ ID NO:3.

[0097] Table 3

[0098]

[0099]

[0100]

[0101] For the purposes of the present invention, the "sequence identity" between two amino acid sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol. 48:443-453) implemented in the Needle program (preferably version 3.0.0 or later) of the EMBOSS software package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, Trends Genet. 16:276-277). The optional parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5 and the EBLOSUM62 (EMBOSS version of BLOSUM62) substitution matrix. The output of Needle marked "longest identity" (obtained using the nobrief option) is used as the percentage identity and is calculated as follows:

[0102] (Number of identical residues x 100) / (length of alignment - total number of gaps in the alignment)

[0103] For the purposes of the present invention, the sequence identity between two deoxyribonucleotide sequences is determined using the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, ibid.) implemented in the Needle program (preferably version 3.0.0 or later) of the EMBOSS software package (EMBOSS: The European Molecular Biology Open Software Suite, Rice et al., 2000, ibid.). The optional parameters used are a gap opening penalty of 10, a gap extension penalty of 0.5 and the EDNAFULL (EMBOSS version of NCBI NUC4.4) substitution matrix. The output of Needle marked "most identity" (obtained using the -nobrief option) is used as the percentage identity and is calculated as follows:

[0104] (Number of identical deoxyribonucleotides x 100) / (length of alignment - total number of gaps in the alignment).

[0105] In one embodiment, the selection step comprises determining whether a bacterial strain contains a manganese transporter having at least 55%, such as at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the sequence of any one of SEQ ID NOs: 1-3. This determination can be based on sequencing the bacterial strain or performing a BLAST search in a known sequence database.

[0106] The manganese scavengers used in the examples of the present invention have a manganese transporter encoding SEQ ID NOs: 1-3 or a functional variant thereof.

[0107] In other embodiments, the present invention provides a method for inhibiting or delaying fungal growth in a product, comprising the steps of:

[0108] - selecting one or more bacterial strains as manganese scavengers, and

[0109] - reducing the free manganese in the product to a concentration in the product below about 0.01 ppm by adding the manganese scavenger,

[0110] wherein the selection step comprises measuring the manganese uptake activity of one or more bacterial strains.

[0111] Manganese uptake activity can be measured using conventional methods known in the art, see, for example, Kehres et al. "The NRAMP proteins of Salmonella typhimurium and Escherichia coli are selective manganese transporters involved in the response to reactive oxygen." Molecular microbiology 36.5 (2000): 1085-1100.

[0112] For fermented foods such as fermented dairy products, the manganese scavenger is preferably Lactobacillus. Different manganese transporter families are present in Lactobacillus, and often multiple homologs of these manganese transporter families are also present. In Figure 11 the inventors provided a phylogenetic tree that outlines the phylogeny of the MntH family of manganese transporters in Lactobacillus species. As shown, manganese transporters can be found in species of the genus Lactobacillus. In addition to species of the genus Lactobacillus, the MntH transporter family can also be found in other bacteria. The tree was constructed using mafft v.7 by aligning Lactobacillus MntH protein sequences, and the phylogeny was inferred by neighbor-joining clustering.

[0113] In a preferred embodiment, the manganese scavenger is a bacterial strain selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus salivarius, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus fermentum, Lactobacillus sakei, Lactobacillus reuteri, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus alimentarius, and Pediococcus acidilactici.

[0114] On the other hand, this transporter does not seem to be present in Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus delbrueckii subsp. bulgaricus, making them less suitable for removing free manganese.

[0115] According to a preferred embodiment of the invention, the method comprises the step of selecting one or more bacterial strains that are determined to be free of superoxide dismutase, preferably free of manganese superoxide dismutase.

[0116] Superoxide dismutase, such as manganese superoxide dismutase, has been studied and is particularly described, for example, in Kehres et al., "Emerging themes in manganese transport, biochemistry and pathogenesis in bacteria." FEMS microbiology reviews 27.2-3 (2003): 263-290, Culotta V.C “Superoxide dismutase, oxidative stress, and cell metabolism” Curr. Top. Cell Regul. 36, 117–132 (2000), or Whittaker J.W “Manganese superoxide dismutase” Met. Ions Biol. Syst. 37, 587–611 (2000).

[0117] In the context of the present invention, the term "free of" means that the genome of one or more bacterial strains does not present a gene encoding superoxide dismutase, or even if the genome of one or more bacterial strains presents a gene encoding superoxide dismutase, this gene is not expressed by the one or more bacterial strains.

[0118] Product

[0119] In some embodiments, the product is a food, cosmetic, health product or pharmaceutical product. "Food" and "food product" have their common meanings. "Food product" means any food or feed product suitable for human or animal consumption. Food products can be fresh or perishable foods, as well as stored or processed foods. Food products include, but are not limited to, fruits and vegetables, including derived products, grains and grain-derived products, dairy products, meat, poultry and seafood. More preferably, the food product is a meat product or a dairy product, such as yogurt, tvarog, sour cream, cheese, etc.

[0120] However, it should be noted that in the context of the present invention, the terms "product" and "food product" in the present invention do in fact relate to water per se. Although manganese is essential for human nutrition, according to the United States Environmental Protection Agency (EPA), it is generally considered that in water, manganese is unhealthy for humans. Therefore, sometimes drinking water or wastewater is treated for purification and health purposes to remove excessive manganese, which is not relevant to the spirit of the present invention.

[0121] The present invention is particularly applicable to food products having a medium to high water activity. Water activity (a w ) determines the viability and function of microorganisms. Water activity or a w is the partial vapor pressure of water in a substance divided by the partial vapor pressure of water in the standard state. In the field of food science, the standard state is usually defined as the partial vapor pressure of pure water at the same temperature. Using this specific definition, the water activity of pure distilled water is exactly 1.

[0122] The main food product types that are prone to fungal spoilage are dairy products having a medium to high water activity, such as yogurt, cream, butter, cheese, etc. However, it is also foreseeable that the present invention is applicable to food products having a lower water activity, such as processed meat, grains, nuts, spices, milk powder, jerky and fermented meat.

[0123] In a preferred embodiment, the water activity (a w)Less than 0.98, such as less than about 0.97, less than about 0.96, less than about 0.95, less than about 0.94, less than about 0.93, less than about 0.92, less than about 0.91, less than about 0.90, less than about 0.89, less than about 0.88, less than about 0.87, less than about 0.86, less than about 0.85, less than about 0.84, less than about 0.83, less than about 0.82, less than about 0.81, less than about 0.80, less than about 0.79, less than about 0.78, less than about 0.77, less than about 0.76, less than about 0.75, less than about 0.74, less than about 0.73, less than about 0.72, less than about 0.71, less than about 0.70 or lower.

[0124] In some embodiments, the product is one having a water activity (a w ) of from about 0.70 to about 0.98, such as from about 0.75 to about 0.97, such as from about 0.80 to about 0.96, such as from about 0.85 to about 0.95.

[0125] Methods for measuring water activity are known in the art, for example, as described in Fontana Jr, Anthony J. "Measurement of water activity, moisture sorption isotherms, and moisture content of foods." Water activity in foods: Fundamentals and applications (2007): 155 - 173.

[0126] In one embodiment, the steps described herein are carried out to inhibit or delay the growth of fungi in fermented foods. Fermented foods are foods produced or preserved by the action of microorganisms. Fermentation refers to the conversion of carbohydrates into alcohols or acids by the action of microorganisms. Fermentation generally refers to the fermentation of sugars to alcohol using yeast. However, it may also involve the conversion of lactose to lactic acid. For example, fermentation can be used to make foods such as yogurt, cheese, salami, sauerkraut, kimchi, pickles, etc.

[0127] In one embodiment, the food is a product of lactic acid fermentation, i.e., a product prepared by fermentation with lactic acid bacteria (LAB). "Lactic acid bacteria" refers to Gram - positive, microaerophilic or anaerobic bacteria that ferment sugars to produce acids, including lactic acid as the main acid produced. The pH of the food is typically from about 3.5 to about 6.5, such as from about 4 to about 6, such as from about 4.5 to about 5.5, such as about 5.

[0128] The present invention is particularly useful in inhibiting or delaying the growth of fungi in dairy products. In such products, contamination by yeasts and molds is common and limits the shelf life of such products. In addition to milk, "dairy products" also include products derived from milk, such as cream, ice cream, butter, cheese, and yogurt, as well as by-products such as whey and casein, and any ready-to-eat food containing milk or milk components as a major ingredient, such as formula milk. In a preferred embodiment, the dairy product is a fermented dairy product. The term "milk" should be understood as the milk secretion obtained by milking any mammal such as cows, sheep, goats, buffalo, or camels. In a preferred embodiment, the milk is cow's milk. The term also includes protein / fat solutions prepared from plant materials, such as soy milk.

[0129] The manganese content in milk varies depending on the animal producing the milk, the feed, and the season. Generally, the concentration of manganese present in dairy products is at least 0.03 ppm, for example, at least 0.08 ppm for skim milk and at least 0.1 ppm for whole milk. According to the discovery of the present inventors, reducing the amount of manganese in such products or products prepared therefrom will make them more resistant to spoilage.

[0130] In one embodiment, the food is a product prepared by fermentation with thermophilic organisms, i.e., thermophilic fermented food. The term "thermophilic organisms" refers to microorganisms that grow best at temperatures above 43°C. The most industrially useful thermophilic bacteria include species of the genus Streptococcus and species of the genus Lactobacillus. The term "thermophilic fermentation" as used herein refers to fermentation at a temperature above about 35°C, such as between about 35°C and about 45°C. "Thermophilic fermented food" refers to fermented food prepared by thermophilic fermentation with a thermophilic starter culture. Such foods include, for example, yogurt, skyr, labneh, lassi, ayran, and doogh.

[0131] In one embodiment, the food is a product prepared by fermentation with mesophilic organisms, i.e., mesophilic fermented food. The term "mesophilic organisms" refers to microorganisms that grow best at moderate temperatures (15°C - 40°C). The most industrially useful mesophilic bacteria include species of the genus Lactococcus and species of the genus Leuconostoc. The term "mesophilic fermentation" as used herein refers to fermentation at a temperature between about 22°C and about 35°C. "Mesophilic fermented food" refers to fermented food prepared by mesophilic fermentation with a mesophilic starter culture. Such foods include, for example, buttermilk, acid milk, fermented milk, smetana, sour cream, and fresh cheeses such as quark, tvarog, and cream cheese.

[0132] Preparation of fermented products

[0133] The methods disclosed herein are particularly useful for inhibiting or delaying the growth of yeasts and / or molds in fermented dairy products, such as thermophilic and mesophilic fermented dairy products such as yogurt products. The term "fermented dairy product" is a term commonly according to relevant official regulations, and the standards are well known in the art. For example, a symbiotic culture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus is used as a starter culture for yogurt, while Lactobacillus acidophilus is used to prepare acidophilus milk. Other mesophilic lactic acid bacteria are used to produce quark cheese or fromage frais.

[0134] The expression "fermented dairy product" refers to a food or feed product, wherein the preparation of the food or feed product involves the fermentation of a milk matrix with lactic acid bacteria. As used herein, "fermented dairy products" include, but are not limited to, products such as thermophilic fermented dairy products (e.g., yogurt) and mesophilic fermented dairy products (e.g., sour cream and buttermilk, as well as fermented whey, quark cheese and fromage frais). Fermented dairy products also include cheeses, such as continental type cheese, fresh cheese, soft cheese, cheddar cheese, mascarpone, pasta filata, mozzarella, provolone, white brine cheese, pizza cheese, feta, brie, camembert, cottage cheese, Edam, gouda, Tilsiter, Havarti or Emmental, Swiss cheese, and Maasdamer.

[0135] The term "yogurt" has its usual meaning and is generally defined according to relevant official regulations, and the standards are well known in the art. The starter culture for preparing yogurt contains at least one strain of Lactobacillus delbrueckii subsp. bulgaricus and at least one strain of Streptococcus thermophilus. Interestingly, manganese transporters are absent in Lactobacillus delbrueckii subsp. bulgaricus and exhibit only low expression in Streptococcus thermophilus, and these two strains are present in the starter culture of yogurt, making them particularly vulnerable to fungal spoilage. Therefore, in order to remove the free manganese present in yogurt, other bacterial strains are preferably included.

[0136] Preferably, the free manganese in the fermented product is reduced to a concentration of less than about 0.005 ppm.

[0137] In the food processing process, chemical preservatives have traditionally been used to avoid fungal spoilage. However, in view of the strong social demand for less processed and preservative-free foods, the present invention provides an effective solution for controlling the growth of yeast and mold by reducing the manganese concentration using a biological manganese scavenger.

[0138] When using a biological scavenger, considering the examples provided by the present invention and the nature of the food, such as water activity, nutrients, the content of naturally occurring manganese, shelf life, storage conditions, packaging, etc., those skilled in the art can adjust various parameters, such as pH, temperature, and the amount of manganese scavenger or bacteria to achieve the desired results.

[0139] It is preferred to package products with a reduced free manganese concentration to further limit contact with yeast and mold. It is also preferred to store the products at a low temperature (below 15 °C) to help extend the shelf life.

[0140] For fermented foods, manganese-removing bacteria can be added before fermentation, at the start of fermentation, or during fermentation. Depending on the selected parameters, the step of reducing the manganese content to a preferred level may take several hours, such as at least 5 hours, such as at least 10 hours, such as at least 15 hours, such as at least 20 hours, such as at least 1 day, 2 days, 3 days or longer. Those skilled in the art will be able to select appropriate parameters according to the products that need to inhibit or delay fungi.

[0141] The present invention provides a method for preparing fermented foods, including adding a starter culture and a manganese scavenger to a food substrate, and fermenting the substrate for a period of time until the target pH is reached. The manganese scavenger is preferably a lactic acid bacteria strain.

[0142] The term "food substrate" used herein refers to the substrate to be fermented.

[0143] To produce fermented dairy products, the food substrate is a milk substrate. The "milk substrate" is widely used in the present invention to represent a milk-based or milk component-based composition that can be used as a medium for the growth and fermentation of starter cultures. "Milk" generally refers to milk secretions obtained by milking any mammal such as cows, sheep, goats, buffaloes, or camels. The milk substrate can be obtained from any unprocessed and / or processed milk material and from reconstituted milk powder. The milk substrate can also be plant-based, i.e., prepared from plant materials such as soy milk. Preferably, the milk substrate is prepared from milk or milk components from cows.

[0144] The milk matrix includes a solution / suspension of any milk or milk-like product containing proteins, such as whole milk or low-fat milk, skim milk, buttermilk, reconstituted milk powder, condensed milk, dried milk.

[0145] Depending on consumer needs, the lactose in the milk base can also be reduced. Lactose-reduced milk can be produced according to any method known in the art, including hydrolysis of lactose to glucose and galactose by lactase, or by nanofiltration, electrodialysis, ion-exchange chromatography, and centrifugation.

[0146] To ferment the milk matrix, a starter culture is added. The term "starter" or "starter culture" as used herein refers to a culture of one or more food-grade microorganisms responsible for acidifying the milk matrix, particularly a culture of lactic acid bacteria.

[0147] A manganese scavenger can be added before fermentation, at the start of fermentation, or during fermentation, either simultaneously with or at different times from the starter culture.

[0148] After adding the starter culture and the manganese scavenger and subjecting the milk matrix to suitable conditions, the fermentation process is started and continued for a period of time. A person of ordinary skill in the art knows how to select suitable process conditions, such as temperature, oxygen, addition of carbohydrates, number and characteristics of microorganisms, and processing time spent. This process may take three hours, four hours, five hours, six hours, or longer.

[0149] These conditions include setting a temperature suitable for the specific strain of the starter culture. For example, when the starter culture contains mesophilic lactic acid bacteria, the temperature can be set to about 30 °C, and if the culture contains thermophilic lactic acid bacteria strains, the temperature is maintained in the range of about 35 °C to 50 °C, such as 40 °C to 45 °C. The setting of the fermentation temperature also depends on the enzyme added to the fermentation, which can be easily determined by a person of ordinary skill in the art. In a specific embodiment of the present invention, the fermentation temperature is 35 °C to 45 °C, preferably 37 °C to 43 °C, and more preferably 40 °C to 43 °C. In another embodiment, the fermentation temperature is 15 °C to 35 °C, preferably 20 °C to 35 °C, and more preferably 30 °C to 35 °C.

[0150] Any method known in the art can be used to terminate the fermentation. Generally, depending on various parameters of the process, the fermentation can be terminated by making the milk matrix unsuitable for the growth of the strain of the starter culture. For example, when the target pH is reached, the fermentation of the fermented dairy product can be terminated by rapid cooling. It is known that acidification occurs during the fermentation process, which results in the formation of a three-dimensional network composed of casein clusters and chains. The term "target pH" refers to the pH at the end of the fermentation step. The target pH depends on the fermented dairy product to be obtained and can be easily determined by a person of ordinary skill in the art.

[0151] In a specific embodiment of the present invention, fermentation is carried out until a pH of at least 5.2 is reached, for example until a pH of 5.1, 5.0, 4.9, 4.8, 4.7, 4.6, 4.5, 4.4, 4.3, 4.2, 4.1, 4.0, 3.9, 3.8 or 3.7 is reached. Preferably, fermentation is carried out until a target pH of 4.0 - 5.0, more preferably 4.0 - 4.6 is reached. In a preferred embodiment, fermentation is carried out until a target pH below 4.6 is reached.

[0152] In a preferred embodiment, the fermented food is selected from the group consisting of quark cheese, cream cheese, cottage cheese, Greek yogurt, skimmed buttermilk, concentrated yogurt, buttermilk, sour cream, acidophilus milk, fermented milk, kefir, Indian yogurt, salted yogurt, twarog, mint salted yogurt, smetana, Yakult, and dahi.

[0153] In another preferred embodiment, the fermented food is cheese, including European cheese, fresh cheese, soft cheese, cheddar cheese, mascarpone cheese, pastor cheese, mozzarella cheese, mozzarella cheese, white brined cheese, pizza cheese, feta cheese, brie cheese, camembert cheese, cottage cheese, red wave cheese, gouda cheese, telsit cheese, havarti cheese or emmental cheese, Swiss cheese, and marston cheese.

[0154] In another embodiment, the method further includes packaging the food to reduce contact with yeast and mold.

[0155] The present invention includes foods obtained by the methods described herein.

[0156] The product obtained by the present invention is preferably a fermented dairy product, the concentration of free manganese in which is reduced to less than 0.01 ppm after storage for at least two days, for example at least 3 days, at least 4 days, more preferably at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days, and at least 14 days.

[0157] Other features and advantages of the present invention will become apparent by reading the following description in conjunction with the accompanying drawings. All terms used herein have the same meaning as commonly understood by one of ordinary skill in the art, unless otherwise defined. The use of the terms "a", "an", "the", and similar referents in the context of describing the present invention (especially in the context of the appended claims) shall be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by the context. The terms "comprising", "having", "including", and "containing" shall be construed as open-ended terms (i.e., meaning "including but not limited to"), unless otherwise specified. The recitation of numerical ranges herein is merely intended to serve as a shorthand method for individually referring to each separate value falling within the range, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order, unless otherwise indicated herein or clearly contradicted by the context. All exact values provided herein represent corresponding approximations (e.g., all exact exemplary values provided for a particular factor or measurement can also be considered to provide the corresponding approximate measurement, modified by "about" where appropriate). The use of any and all examples or exemplary language (e.g., "such as") provided herein is merely intended to better illustrate the invention and is not intended to limit the scope of the invention. No language in the specification should be construed as indicating any non-claimed element as essential for the practice of the invention.

[0158] Example

[0159] The scope of the invention described and claimed herein is not limited by the specific aspects disclosed herein, as these aspects are intended to illustrate several aspects of the invention. Any equivalent aspects are intended to be within the scope of the invention. Indeed, various modifications of the invention will become apparent to those skilled in the art from the foregoing description and the following examples, in addition to those shown and described herein. Such modifications are also intended to fall within the scope of the appended claims. In case of conflict, the present disclosure, including definitions, will control.

[0160] Example 1: Growth of yeast at different manganese concentrations

[0161] This example demonstrates the requirement of Debaryomyces hansenii for manganese in minimal medium. Two strains of Debaryomyces hansenii isolated from spoiled yogurt (strain 1) and quark cheese (strain 2) were used. The strains were grown in chemically defined media with different manganese concentrations.

[0162] The basal medium contains biotin 2 μg / L, calcium pantothenate 400 μg / L, folic acid 2 μg / L, inositol 2 μg / L, nicotinic acid 400 μg / L, p-aminobenzoic acid 200 μg / L, pyridoxine 400 μg / L, riboflavin 200 μg / L, thiamine 400 μg / L, boric acid 500 μg / L, copper sulfate 40 μg / L, potassium iodide 100 μg / L, ferric chloride 200 μg / L, sodium molybdate 200 μg / L, zinc sulfate 400 μg / L, potassium dihydrogen phosphate 0.5 g / L, dipotassium hydrogen phosphate 0.5 g / L, magnesium sulfate 0.5 g / L, sodium chloride 0.1 g / L, calcium chloride 0.2 g / L, glucose 20 g / L, ammonium sulfate 5 g / L.

[0163] The manganese concentrations used: 6 ppm, 0.6 ppm, 0.06 ppm, 0.006 ppm, 0.0006 ppm, 0.00006 ppm, 0.000006 ppm.

[0164] Two different pHs were tested: pH 6.5 and 4.5.

[0165] The strains were inoculated into 150 μl of different media in 96-well plates. The plates were incubated at 17 °C for several days, and after the yeast strains grew, the absorbance at 600 nm was measured in a plate reader.

[0166] Figure 1 (Strain 1) and Figure 2 (Strain 2) showed the effects of different pHs and different manganese concentrations on the growth of two different Debaryomyces hansenii strains. It can be seen that the growth of Debaryomyces hansenii was inhibited at manganese concentrations below about 0.01 ppm. There was no difference between the two different pH values, indicating that the mechanism was effective within this pH range.

[0167] Example 2: Inhibition of yeast in fermented dairy products

[0168] This example demonstrated the manganese requirement of Debaryomyces hansenii in fermented dairy products.

[0169] Fermented dairy products were prepared with a starter culture (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) or additionally with a manganese scavenger (Lactobacillus rhamnosus and Lactobacillus paracasei). The fermented dairy products were centrifuged (10 minutes at 5000 rpm), and the supernatant was sterile filtered. The supernatant was transferred to sterile 96-well plates (150 μl per well), and manganese was added to the first well to a final concentration of 6 ppm, followed by 10-fold serial dilutions to obtain different manganese concentrations from 6 ppm to 0.000006 ppm. Debaryomyces hansenii was inoculated at approximately 20 cells / well, and the plates were incubated at 17 °C for several days. After the yeast strains grew, the absorbance at 600 nm was measured in a plate reader on the 7th day.

[0170] Figure 3 and 4 showed the growth of Debaryomyces hansenii (strains 1 and 2, respectively). After adding manganese to the aqueous phase, the yeast growth of strain 1 was as Figure 3 shown, while the yeast growth of strain 2 was as Figure 4 shown (hollow circles). The mean and standard deviation of technical replicates are shown for n = 6 (A) and n = 3 (B). For comparison, the yeast growth of the reference yogurt aqueous phase is also shown, in which neither a manganese scavenger nor additional manganese was added (squares). Note that the inherent manganese concentration of this reference was 0.03 ppm. For the hollow circles, the x-axis represents the added manganese concentration, and for the squares (reference yogurt aqueous phase), the x-axis represents the manganese inherent in the aqueous phase.

[0171] These results indicate that the growth of yeast strains in food matrices depends on manganese. The addition of manganese led to growth similar to the reference, thus demonstrating that low manganese concentration is the main limiting factor for yeast growth in fermented dairy products.

[0172] Example 3. Inhibition of Debaryomyces and Rhodoturola

[0173] This example demonstrated the manganese scavenging activity of various bacteria and their inhibitory effects on Debaryomyces and Rhodotorula. The inhibitory effects were evaluated in fermented dairy products with low and high manganese concentrations.

[0174] Table 4 lists the added bacteria and whether they contain manganese transporters.

[0175] Table 4

[0176] Number Species Containing manganese transporter 1 Lactobacillus rhamnosus Yes 2 Lactobacillus rhamnosus Yes 3 Lactobacillus rhamnosus Yes 4 Lactobacillus rhamnosus (ATCC 7469) Yes 5 Lactobacillus salivarius Yes 6 Lactobacillus casei Yes 7 Lactobacillus paracasei Yes 8 Lactobacillus fermentum Yes 9 Lactobacillus sakei (ATCC 15521) Yes 10 Lactobacillus reuteri (ATCC 23272) Yes 11 Lactobacillus plantarum Yes 12 Lactobacillus delbrueckii No 13 Lactobacillus helveticus No 14 Lactobacillus gasseri (ATCC 33323) No 15 Lactobacillus brevis Yes 16 Lactobacillus kefiranofaciens (ATCC 35411) Yes 17 Lactobacillus alimentarius Yes 18 Pediococcus acidilactici (DSM20284) Yes 19 Control (only starter culture)

[0177] The listed bacteria were grown overnight in MRS medium.

[0178] Preparation: Inoculate 10 μl of the overnight culture into 2 ml of milk containing a starter culture (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus). Ferment the milk at 43 °C for approximately 6 hours until a pH of 4.5 is reached. Store the fermented milk in the refrigerator for further use. Transfer 150 μl of the fermented milk in duplicate to each well of a 96-well plate. Add manganese to half of the samples to increase the manganese by 6 ppm, and inoculate approximately 20 cells of Debaryomyces hansenii or Rhodotorula glutinis into all wells. After 4 days, spot a row of dilutions on a selective YGC plate to analyze yeast growth. Yeast growth was measured by visual inspection, giving 0 for no growth and 5 for complete growth. Table 5 shows the mean of two biologically independent experiments for Debaryomyces, while Table 6 shows the mean of two biologically independent experiments for Rhodotorula.

[0179] Table 5 Inhibition of Debaryomyces

[0180]

[0181] Table 6 Inhibition of Rhodotorula

[0182]

[0183]

[0184] The results showed that the manganese-removing strains could be used to inhibit Debaryomyces hansenii and Rhodotorula glutinis, but the inhibitory effect weakened after adding manganese.

[0185] Example 4. Inhibition of Debaryomyces, Saccharomyces, Rhodotorula, Cryptococcus and Torulaspora

[0186] This example evaluated the differences in yeast growth in the aqueous phase of fermented milk prepared with a starter culture with and without a manganese scavenger.

[0187] Table 7 lists the manganese scavengers used:

[0188] Table 7

[0189]

[0190] This example showed yeast growth at two different manganese concentrations. The inhibitory effects on 6 different yeasts in fermented dairy products with low and high manganese concentrations were evaluated.

[0191] The lipid-reduced (1.5% w / v) homogenized milk was heat-treated at 90 ± 1 °C for 20 minutes and then immediately cooled. A commercial starter culture (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) was inoculated at 0.02% (v / w) in 3 L buckets. One bucket was inoculated with a manganese scavenger at a total concentration of 100 U / T, and one bucket was used as a reference and only inoculated with the starter culture. All buckets were incubated in a water bath at 43 ± 1 °C and fermented under these conditions until a pH of 4.60 ± 0.1 was reached. The fermented dairy product was divided into 200 mL bottles and cooled.

[0192] Then the fermented dairy product was centrifuged (10 minutes at 5000 rpm), and the supernatant was sterile filtered. The supernatant was transferred to a sterile 96-well plate (150 μl per well), and manganese was added to half of the wells to increase the manganese by 6 ppm. Six different yeasts were selected, and approximately 20 cells were inoculated into each well and grown at 17 °C for 7 days. The milk fermented only with the starter culture (reference) was used as a control. Growth was determined by measuring the absorbance at 600 nm.

[0193] Tables 8 - 13 show the mean values and standard deviations of at least 5 replicates.

[0194] Table 8 Inhibition of Debaryomyces hansenii (strain 1):

[0195]

[0196] Table 9 Inhibition of Debaryomyces hansenii (strain 2)

[0197]

[0198] Table 10 Inhibition of Saccharomyces cerevisiae

[0199]

[0200]

[0201] Table 11 Inhibition of Rhodotorula glutinis

[0202]

[0203] Table 12 Inhibition of Cryptococcus laurentii

[0204]

[0205] Table 13 Inhibition of Torulaspora delbrueckii

[0206]

[0207] The results showed that manganese-clearing bacteria were able to inhibit strains of the genera Debaryomyces, Saccharomyces, Rhodotorula, Cryptococcus, and Torulaspora, and the inhibitory effect was reduced after the addition of manganese.

[0208] Example 5. Inhibition of Debaryomyces hansenii

[0209] This example evaluated and tested the effect of different manganese concentrations on fermented milk prepared with a starter culture with or without a manganese scavenger.

[0210] Table 14 lists the manganese scavengers used:

[0211] Table 14

[0212]

[0213] Prepare fermented dairy products as in Example 4.

[0214] Transfer 150 μl of fermented milk in duplicate or triplicate to each well of a 96-well plate. Perform serial dilutions to result in different added manganese concentrations ranging from 6 ppm to 0.000006 ppm. Inoculate all wells with approximately 20 cells of Debaryomyces hansenii (strain 2), and incubate the plate at 17 °C for 5 days. Thereafter, spot 10 μl of a 1000-fold dilution prepared in saline peptone onto a selective YGC plate to analyze yeast growth.

[0215] Figure 5 Shows the growth of Debaryomyces hansenii in fermented milk prepared with (1, 2, and 3) and without (REF (reference)) scavengers after adding different manganese concentrations: 6 ppm (top row), 0.6 ppm (second row), 0.06 ppm (third row), 0.006 ppm (fourth row), 0.0006 ppm (fifth row), 0.00006 ppm (sixth row), 0.000006 ppm (seventh row), and 0 ppm (bottom row).

[0216] As shown, after adding 0.6 ppm, the growth of yeast in fermented milk with scavengers 1 and 2 was inhibited (compare the third row and the second row); in the presence of scavenger 3, adding 0.006 ppm did cause some yeast growth.

[0217] Example 6. Inhibition of yeast in fermented dairy products with different manganese contents

[0218] The effect of manganese on the inhibition of different molds was evaluated. An agar assay similar to the manufacturing method and production of fermented dairy products was used. Lactobacillus rhamnosus and Lactobacillus paracasei were used together as manganese scavengers.

[0219] Preparation of fermented milk samples:

[0220] The fat-reduced (1.5% w / v) homogenized milk was heat-treated at 90 ± 1 °C for 20 minutes and then immediately cooled. A commercial starter culture (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) was inoculated at 0.02% (v / w) in 3 L buckets. One bucket was inoculated with a manganese scavenger at a total concentration of 100 U / T, and one bucket was used as a reference and inoculated only with the starter culture. All buckets were incubated in a water bath at 43 ± 1 °C and fermented under these conditions until a pH of 4.60 ± 0.1 was reached. The fermented dairy product was dispensed into 200 mL bottles and cooled. The manganese concentration already present in the reference product was previously determined to be approximately 0.03 ppm, while the manganese concentration in the product with the scavenging strain was below the detection limit of 0.003 ppm.

[0221] Adding manganese:

[0222] In the presence and absence of manganese scavengers, different concentrations of manganese were added to the fermented dairy product to increase the manganese content (0, 0.006, and 6 ppm of manganese in the reference product, and 0, 0.000006, 0.00006, 0.0006, 0.006, 0.06, 0.6, and 6 ppm of manganese).

[0223] Heat all the fermented milk samples to a temperature of 40 °C and add 40 ml of a 5% sterile agar solution that has been melted and cooled to 60 °C. Then pour this solution of fermented milk and agar into a sterile Petri dish and dry the plate in a LAF workbench for 30 minutes.

[0224] Attack test using yeast:

[0225] At 10 3 、10 2 and 10 1 CFU / dot concentration, spot three target contaminants, including two strains of Debaryomyces hansenii (strain 1 and strain 2) and one strain of Cryptococcus longirostris. Incubate the plates at 7 ± 1 °C and regularly check for yeast growth.

[0226] Results:

[0227] The results of the yeast agar assay are shown in Figure 6 and Figure 6 show the growth of three different yeasts on plates prepared from milk fermented with a separate starter culture (reference, top row) or together with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were added. At three different concentrations: 1x10 3 cfu / dot (top row on the plate), 1x10 2 cfu / dot (middle row on the plate) and 1x10 1 cfu / dot (bottom row on the plate), three target contaminants were added (Column A: Debaryomyces hansenii (strain 2), Column B: Cryptococcus longirostris, Column C: Debaryomyces hansenii (strain 1).

[0228] It can be seen from Figure 6 that the tested yeasts grew well on agar plates prepared from milk fermented with only the starter culture (reference). However, when a manganese scavenger was present during the milk fermentation, the growth halos of all the yeasts were delayed.

[0229] When the manganese content was as high as 0.0006 ppm, the scavenger maintained a high inhibitory activity against all three yeasts. When the manganese content was 0.006 ppm to 0.6 ppm, the inhibitory activity of the scavenger against Cryptococcus longirostris decreased. A manganese concentration of 6 ppm seemed to inhibit the growth of Cryptococcus longirostris. The manganese scavenger inhibited Debaryomyces hansenii (strain 1) when the manganese content was as high as 0.006 ppm. When the manganese content was 0.06 ppm or higher, the manganese scavenger lost its inhibitory activity against Debaryomyces hansenii (strain 1). The manganese scavenger inhibited Debaryomyces hansenii (strain 2) when the manganese content was as high as 0.6 ppm, and the activity was lost when the manganese content was 6 ppm.

[0230] Example 7: Inhibition of molds in fermented dairy products with different manganese contents

[0231] Prepare fermented dairy product samples with different manganese contents as described in Example 6.

[0232] Attack test using molds:

[0233] Figure 7 Shows the growth of three different molds on plates prepared from milk fermented with a starter culture alone (reference, top row) or additionally with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were further added. Three target contaminants (A: Penicillium brevicompactum, B: Penicillium crustosum, and C: Penicillium dissocians) were added at a concentration of 500 spores / spot. The plates were incubated at 7 ± 1 °C for 25 days.

[0234] Figure 8 Shows the growth of three different molds on plates prepared from milk fermented with a starter culture alone (reference, top row) or additionally with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were further added. Three target contaminants (A: Penicillium brevicompactum, B: Penicillium crustosum, and C: Penicillium dissocians) were added at a concentration of 500 spores / spot. The plates were incubated at 22 ± 1 °C for 8 days.

[0235] Figure 9 Shows the growth of three different molds on plates prepared from milk fermented with a starter culture alone (reference, top row) or additionally with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were further added. Three target contaminants (A: Penicillium carneum, B: Penicillium vinaceum, and C: Penicillium roqueforti) were added at a concentration of 500 spores / spot. The plates were incubated at 7 ± 1 °C for 25 days.

[0236] Figure 10 Shows the growth of three different molds on plates prepared from milk fermented with a starter culture alone (reference, top row) or additionally with a manganese scavenger (bottom row). As shown above the picture, different manganese concentrations were further added. Three target contaminants (A: Penicillium carneum, B: Penicillium vinaceum, and C: Penicillium roqueforti) were added at a concentration of 500 spores / spot. The plates were incubated at 22 ± 1 °C for 8 days.

[0237] All the tested molds grew well on agar plates made from milk fermented with the starter culture alone (reference). However, when a manganese scavenger was present during the milk fermentation, the growth halos of all the tested molds were delayed. The inhibitory effect was stronger at lower temperatures ( Figure 7 and 9 ).

[0238] The manganese scavenger maintained high inhibitory activity against all molds at additional manganese contents up to 0.006 ppm. At manganese contents of 0.06 ppm and above, the inhibitory activity of the manganese scavenger decreased. At 6 ppm, the manganese scavenger lost most of its inhibitory activity against sensitive organisms ( Figure 7 and 8 ), and at 0.06 ppm it lost most of its inhibitory activity against robust molds ( Figure 9 and 10 ).

[0239] The added manganese concentration that weakens the inhibition found in this assay is higher compared to the concentrations found in the aqueous phase because of the continuous absorption of the live metabolically active manganese scavenger in the fermented dairy product.

[0240] Example 8 Inhibition of Debaryomyces hansenii in aqueous phase and different concentrations of chemical chelating materials

[0241] In Example 8, two concentrations of manganese were used: 6 ppm and 0.6 ppm. It has been shown that a concentration of 6 ppm has an inhibitory / toxic effect on yeast growth, while 0.6 ppm is used as the standard concentration sufficient to abolish manganese deficiency caused by one or more bacterial strains acting as manganese scavengers.

[0242] Figure 12 and 13 It was shown that chemical chelating materials, such as EDTA, have an inhibitory effect on the growth of Debaryomyces hansenii in the aqueous phase.

[0243] Figure 12 It was shown that when faced with EDTA at a concentration of 0.05 mg / ml, Debaryomyces hansenii cells stopped growing. Thus, EDTA exhibited an inhibitory effect at a concentration of 0.05 mg / ml.

[0244] Figure 13 It was shown that at a concentration of 0.05 mg / ml EDTA and in the absence of manganese (0.6 ppm), Debaryomyces hansenii stopped growing. Thus, under these conditions, EDTA has an inhibitory effect on Debaryomyces hansenii. Adding 0.6 ppm of manganese and 0.05 mg / ml of EDTA restored the growth of Debaryomyces hansenii, while excess manganese eliminated the effect of EDTA.

[0245] Example 9. Inhibition of Debaryomyces hansenii in yogurt

[0246] In yogurt, the inhibition of a manganese scavenger selected from one or more bacterial strains (scavenger 1) can be reproduced by adding up to 0.89 mg / ml of the manganese scavenger EDTA to the yogurt. At said concentration, the inhibition exhibited by the reference yogurt was similar to that observed when adding scavenger 1. Figure 14The negative control of the bottom row (without added EDTA) showed normal inhibition in the presence of manganese scavenger 1 and restored yeast growth upon addition of manganese.

[0247] Example 9 shows that chemical chelating materials, such as EDTA, have the same effect as manganese scavengers selected from one or more of the bacteria disclosed herein ( Figure 14 ). Thus, Example 9 demonstrates that EDTA has the same effect as one or more bacterial strains disclosed herein as manganese scavengers.

[0248] When present in a given product, such as food, manganese scavengers, chemical chelating materials, and / or biological materials, such as one or more bacterial strains, cause manganese depletion against spoilage fungi, thereby inhibiting fungal growth. Sequence Listing <110> Chr. Hansen A / S <120> Inhibition of Fungal Growth by Manganese Depletion <130> P6438 <150> EP18168109.9 <151> 2018-04-19 <160> 25 <170> BiSSAP 1.3.6 <210> 1 <211> 458 <212> PRT <213> Lactobacillus <400> 1 Met Ala Ser Glu Asp Lys Lys Ser Lys Arg Glu His Ile Ile His Phe 1 5 10 15 Glu Asp Thr Pro Ser Lys Ser Leu Asp Glu Val Asn Gly Ser Val Glu 20 25 30 Val Pro His Asn Ala Gly Phe Trp Lys Thr Leu Ala Ala Tyr Thr Gly 35 40 45 Pro Gly Ile Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ile 50 55 60 Thr Ser Ile Ala Gly Gly Ala Ser Phe Lys Tyr Ser Leu Leu Ser Val 65 70 75 80 Ile Leu Ile Ser Ser Leu Ile Ala Met Leu Leu Gln Ala Met Ala Ala 85 90 95 Arg Leu Gly Ile Val Thr Gly Arg Asp Leu Ala Gln Met Thr Arg Asp 100 105 110 His Thr Ser Lys Ala Met Gly Gly Phe Leu Trp Val Ile Thr Glu Leu 115 120 125 Ala Ile Met Ala Thr Asp Ile Ala Glu Ile Ile Gly Ser Ala Ile Ala 130 135 140 Leu Lys Leu Leu Phe Asn Met Pro Leu Ile Val Gly Ile Ile Ile Thr 145 150 155 160 Thr Ala Asp Val Leu Ile Leu Leu Leu Leu Met Arg Leu Gly Phe Arg 165 170 175 Lys Ile Glu Ala Val Val Ala Thr Leu Val Leu Val Ile Leu Leu Val 180 185 190 Phe Ala Tyr Glu Val Ile Leu Ala Gln Pro Asn Val Pro Glu Leu Leu 195 200 205 Lys Gly Tyr Leu Pro His Ala Asp Ile Val Thr Asn Lys Ser Met Leu 210 215 220 Tyr Leu Ser Leu Gly Ile Val Gly Ala Thr Val Met Pro His Asp Leu 225 230 235 240 Phe Leu Gly Ser Ser Ile Ser Gln Thr Arg Lys Ile Asp Arg Thr Lys 245 250 255 His Glu Glu Val Lys Lys Ala Ile Lys Phe Ser Thr Ile Asp Ser Asn 260 265 270 Leu Gln Leu Thr Met Ala Phe Ile Val Asn Ser Leu Leu Leu Ile Leu 275 280 285 Gly Ala Ala Leu Phe Phe Gly Thr Ser Ser Ser Val Gly Arg Phe Val 290 295 300 Asp Leu Phe Asn Ala Leu Ser Asn Ser Gln Ile Val Gly Ala Ile Ala 305 310 315 320 Ser Pro Met Leu Ser Met Leu Phe Ala Val Ala Leu Leu Ala Ser Gly 325 330 335 Gln Ser Ser Thr Ile Thr Gly Thr Leu Ala Gly Gln Ile Ile Met Glu 340 345 350 Gly Phe Ile His Leu Lys Met Pro Leu Trp Ala Gln Arg Leu Leu Thr 355 360 365 Arg Leu Met Ser Val Thr Pro Val Leu Ile Phe Ala Ile Tyr Tyr His 370 375 380 Gly Asn Glu Ala Lys Ile Glu Asn Leu Leu Thr Phe Ser Gln Val Phe 385 390 395 400 Leu Ser Ile Ala Leu Pro Phe Ala Val Ile Pro Leu Val Leu Tyr Thr 405 410 415 Ser Asp Lys Lys Ile Met Gly Glu Phe Ala Asn Arg Ala Trp Val Lys 420 425 430 Trp Thr Ala Trp Phe Ile Ser Gly Val Leu Ile Ile Leu Asn Leu Tyr 435 440 445 Leu Ile Ala Gln Thr Leu Gly Phe Val Lys 450 455 <210> 2 <211> 450 <212> PRT <213> Lactobacillus <400> 2 Met Ala Arg Pro Asp Glu Arg Leu Thr Val Gln Arg Glu Lys Arg Ser 1 5 10 15 Leu Asp Asp Ile Asn Arg Ser Val Gln Val Pro Ser Val Tyr Glu Ser 20 25 30 Ser Phe Phe Gln Lys Phe Leu Ala Tyr Ser Gly Pro Gly Ala Leu Val 35 40 45 Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Leu Thr Ala Leu Glu Gly 50 55 60 Gly Ser Arg Tyr His Tyr Ala Leu Leu Ser Val Leu Leu Met Ser Ile 65 70 75 80 Leu Val Ala Met Phe Met Gln Thr Leu Ala Ile Lys Leu Gly Val Val 85 90 95 Ala Arg Leu Asp Leu Ala Gln Ala Ile Ala Ala Phe Ile Pro Asn Trp 100 105 110 Ser Arg Ile Cys Leu Trp Leu Ile Asn Glu Ala Ala Met Met Ala Thr 115 120 125 Asp Met Thr Gly Val Val Gly Thr Ala Ile Ala Leu Lys Leu Leu Phe 130 135 140 Gly Leu Pro Leu Met Trp Gly Met Leu Leu Thr Ile Ala Asp Val Leu 145 150 155 160 Val Val Leu Leu Phe Leu Arg Phe Gly Ile Arg Arg Ile Glu Leu Ile 165 170 175 Val Leu Val Ser Ile Leu Thr Val Gly Ile Ile Phe Gly Ile Glu Val 180 185 190 Ala Arg Ala Asp Pro Ser Ile Gly Gly Ile Ala Gly Gly Phe Val Pro 195 200 205 His Thr Asp Ile Leu Thr Asn His Gly Met Leu Leu Leu Ser Leu Gly 210 215 220 Ile Met Gly Ala Thr Ile Met Pro His Asn Ile Tyr Leu His Ser Ser 225 230 235 240 Leu Ala Gln Ser Arg Lys Tyr Asp Glu His Ile Pro Ala Gln Val Thr 245 250 255 Glu Ala Leu Arg Phe Gly Lys Trp Asp Ser Asn Val His Leu Val Ala 260 265 270 Ala Phe Leu Ile Asn Ala Leu Leu Leu Ile Leu Gly Ala Ala Leu Phe 275 280 285 Tyr Gly Val Gly Gly His Val Thr Ala Phe Gln Gly Ala Tyr Asn Gly 290 295 300 Leu Lys Asn Pro Met Ile Val Gly Gly Leu Ala Ser Pro Leu Met Ser 305 310 315 320 Thr Leu Phe Ala Phe Ala Leu Leu Ile Thr Gly Leu Ile Ser Ser Ile 325 330 335 Ala Ser Thr Leu Ala Gly Gln Ile Val Met Glu Gly Tyr Leu Asn Ile 340 345 350 Arg Met Pro Leu Trp Glu Arg Arg Leu Leu Thr Arg Leu Val Thr Leu 355 360 365 Ile Pro Ile Met Val Ile Gly Phe Met Ile Gly Phe Ser Glu His Asn 370 375 380 Phe Glu Gln Val Ile Val Tyr Ala Gln Val Ser Leu Ser Ile Ala Leu 385 390 395 400 Pro Phe Thr Leu Phe Pro Leu Val Ala Leu Thr Asn Arg Arg Asp Leu 405 410 415 Met Gly Ile His Val Asn Ser Gln Leu Val Arg Trp Val Gly Tyr Phe 420 425 430 Leu Thr Gly Val Ile Thr Val Leu Asn Ile Gln Leu Ala Ile Ser Val 435 440 445 Phe Val 450 <210> 3 <211> 535 <212> PRT <213> Lactobacillus <400> 3 Met Ser Asp Asp His Lys Lys Arg His Pro Ile Lys Leu Ile Gln Tyr 1 5 10 15 Ala Asn Gly Pro Ser Leu Glu Glu Ile Asn Gly Thr Val Glu Val Pro 20 25 30 His Gly Lys Gly Phe Trp Arg Thr Leu Phe Ala Tyr Ser Gly Pro Gly 35 40 45 Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr Ser 50 55 60 Ile Thr Gly Gly Gln Asn Phe Gln Tyr Leu Leu Ile Ser Val Ile Leu 65 70 75 80 Met Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys Leu 85 90 95 Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg Thr 100 105 110 Ser Lys Lys Leu Gly Ile Val Leu Trp Ile Leu Thr Glu Leu Ala Ile 115 120 125 Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu Tyr 130 135 140 Leu Leu Phe His Ile Pro Leu Val Ile Ala Val Leu Val Thr Val Leu 145 150 155 160 Asp Val Leu Val Leu Leu Leu Leu Thr Lys Ile Gly Phe Arg Lys Ile 165 170 175 Glu Ala Ile Val Val Ala Leu Ile Leu Val Ile Leu Leu Val Phe Val 180 185 190 Tyr Gln Val Ala Leu Ser Asp Pro Asn Met Gly Ala Leu Leu Lys Gly 195 200 205 Phe Ile Pro Thr Gly Glu Thr Phe Ala Ser Ser Pro Ser Ile Asn Gly 210 215 220 Met Ser Pro Ile Gln Gly Ala Leu Gly Ile Ile Gly Ala Thr Val Met 225 230 235 240 Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln Thr Arg Lys Ile 245 250 255 Asp Tyr Lys Asn Pro Asp Asp Val Ala Gln Ala Val Lys Phe Ser Ala 260 265 270 Trp Asp Ser Asn Ile Gln Leu Ser Phe Ala Phe Val Val Asn Cys Leu 275 280 285 Leu Leu Val Met Gly Val Ala Val Phe Lys Ser Gly Ala Val Lys Asp 290 295 300 Pro Ser Phe Phe Gly Leu Phe Gln Ala Leu Ser Asp Ser Ser Thr Leu 305 310 315 320 Ser Asn Gly Val Leu Ile Ala Val Ala Lys Ser Gly Ile Leu Ser Ile 325 330 335 Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile Thr 340 345 350 Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Val His Met Lys 355 360 365 Met Pro Leu Trp Ala Arg Arg Leu Val Thr Arg Ile Ile Ser Val Ile 370 375 380 Pro Val Ile Val Cys Val Met Leu Thr Ala Arg Asp Thr Pro Ile Gln 385 390 395 400 Gln His Glu Ala Leu Asn Thr Leu Met Asn Asn Ser Gln Val Phe Leu 405 410 415 Ala Phe Ala Leu Pro Phe Ser Met Leu Pro Leu Leu Met Phe Thr Asn 420 425 430 Ser Lys Val Glu Met Gly Asp Arg Phe Lys Asn Thr Gly Trp Val Lys 435 440 445 Val Leu Gly Trp Ile Ser Val Leu Gly Leu Thr Gly Leu Asn Leu Lys 450 455 460 Gly Leu Pro Asp Ser Ile Ala Gly Phe Phe Gly Asp His Pro Thr Ala 465 470 475 480 Thr Gln Thr Asn Met Ala Asn Ile Ile Ala Ile Val Leu Ile Val Ala 485 490 495 Ile Leu Ala Leu Leu Ala Trp Thr Ile Trp Asp Leu Tyr Lys Gly Asn 500 505 510 Gln Arg Tyr Glu Ala His Leu Ala Ala Val Ala Asp Glu Lys Glu Ala 515 520 525 Lys Ala Asp Val Asp Glu Gln 530 535 <210> 4 <211> 458 <212> PRT <213> Lactobacillus casei <400> 4 Met Ala Ser Glu Asp Lys Lys Ser Lys Arg Glu His Ile Ile His Phe 1 5 10 15 Glu Asp Thr Pro Ser Lys Ser Leu Asp Glu Val Asn Gly Ser Val Glu 20 25 30 Val Pro His Asn Ala Gly Phe Trp Lys Thr Leu Ala Ala Tyr Thr Val 35 40 45 Pro Gly Ile Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ile 50 55 60 Thr Ser Ile Ala Gly Gly Ala Ser Phe Lys Tyr Ser Leu Leu Ser Val 65 70 75 80 Ile Leu Ile Ser Ser Leu Ile Ala Met Leu Leu Gln Ala Met Ala Ala 85 90 95 Arg Leu Gly Ile Val Thr Gly Arg Asp Leu Ala Gln Met Thr Arg Asp 100 105 110 His Thr Ser Lys Ala Met Gly Gly Phe Leu Trp Val Ile Thr Glu Leu 115 120 125 Ala Ile Met Ala Thr Asp Ile Ala Glu Ile Ile Gly Ser Ala Ile Ala 130 135 140 Leu Lys Leu Leu Phe Asn Met Pro Leu Ile Val Gly Ile Ile Ile Thr 145 150 155 160 Thr Ala Asp Val Leu Ile Leu Leu Leu Leu Met Arg Leu Gly Phe Arg 165 170 175 Lys Ile Glu Ala Val Val Ala Thr Leu Val Leu Val Ile Leu Leu Val 180 185 190 Phe Ala Tyr Glu Val Ile Leu Ala Gln Pro Asn Val Pro Glu Leu Leu 195 200 205 Lys Gly Tyr Leu Pro His Ala Asp Ile Val Thr Asn Lys Ser Met Leu 210 215 220 Tyr Leu Ser Leu Gly Ile Val Gly Ala Thr Val Met Pro His Asp Leu 225 230 235 240 Phe Leu Gly Ser Ser Ile Ser Gln Thr Arg Lys Ile Asp Arg Thr Lys 245 250 255 His Glu Glu Val Lys Lys Ala Ile Lys Phe Ser Thr Ile Asp Ser Asn 260 265 270 Leu Gln Leu Thr Met Ala Phe Ile Val Asn Ser Leu Leu Leu Ile Leu 275 280 285 Gly Ala Ala Leu Phe Phe Gly Thr Ser Ser Ser Val Gly Arg Phe Val 290 295 300 Asp Leu Phe Asn Ala Leu Ser Asn Ser Gln Ile Val Gly Ala Ile Ala 305 310 315 320 Ser Pro Met Leu Ser Met Leu Phe Ala Val Ala Leu Leu Ala Ser Gly 325 330 335 Gln Ser Ser Thr Ile Thr Gly Thr Leu Ala Gly Gln Ile Ile Met Glu 340 345 350 Gly Phe Ile His Leu Lys Met Pro Leu Trp Ala Gln Arg Leu Leu Thr 355 360 365 Arg Leu Met Ser Val Thr Pro Val Leu Ile Phe Ala Ile Tyr Tyr His 370 375 380 Gly Asn Glu Ala Lys Ile Glu Asn Leu Leu Thr Phe Ser Gln Val Phe 385 390 395 400 Leu Ser Ile Ala Leu Pro Phe Ala Val Ile Pro Leu Val Leu Tyr Thr 405 410 415 Ser Asp Lys Lys Ile Met Gly Glu Phe Ala Asn Arg Ala Trp Val Lys 420 425 430 Trp Thr Ala Trp Phe Ile Ser Gly Val Leu Ile Ile Leu Asn Leu Tyr 435 440 445 Leu Ile Ala Gln Thr Leu Gly Phe Val Lys 450 455 <210> 5 <211> 459 <212> PRT <213> Lactobacillus brevis <400> 5 Met Lys Asn His Glu Thr Asp Thr Lys Thr Lys His His Met Ile Glu 1 5 10 15 Ser Thr Gly Ser Gly Gln Lys Ser Leu Asp Glu Val Asn Gly Thr Val 20 25 30 Glu Val Pro Gln Asn Ala Gly Phe Trp Arg Thr Leu Met Ala Tyr Thr 35 40 45 Gly Pro Gly Ala Leu Ile Ala Val Gly Tyr Met Asp Pro Gly Asn Trp 50 55 60 Ile Thr Ser Ile Ala Gly Gly Ala Gln Tyr Lys Tyr Thr Leu Leu Thr 65 70 75 80 Val Val Leu Leu Ser Ser Leu Val Ala Met Leu Leu Gln Ala Met Ser 85 90 95 Ala Arg Leu Gly Ile Val Thr Gly Lys Asp Leu Ala Gln Leu Thr Arg 100 105 110 Glu His Thr Gly Lys Arg Thr Gly Phe Ala Leu Trp Ile Ile Thr Glu 115 120 125 Leu Ala Ile Met Ala Thr Asp Ile Ala Glu Ile Ile Gly Ser Ala Ile 130 135 140 Ala Leu Lys Leu Leu Phe Gly Phe Pro Leu Ile Val Gly Ile Ile Ile 145 150 155 160 Thr Ala Met Asp Val Leu Val Leu Leu Val Leu Met Lys Leu Gly Phe 165 170 175 Arg Lys Ile Glu Ala Ile Val Ala Thr Leu Val Ala Val Ile Leu Phe 180 185 190 Val Phe Leu Tyr Glu Val Ile Leu Ala Gln Pro His Met Gly Glu Val 195 200 205 Leu Lys Gly Tyr Leu Pro Ser Ser Thr Val Val Thr Asn His Gly Met 210 215 220 Leu Tyr Leu Ser Leu Gly Ile Val Gly Ala Thr Val Met Pro His Asp 225 230 235 240 Leu Tyr Leu Gly Ser Ser Ile Ser Gln Thr Arg Ser Phe Asp Arg Lys 245 250 255 Asn Arg Lys Ser Val Ala Gln Ala Ile Lys Phe Thr Thr Ile Asp Ser 260 265 270 Asn Ile Gln Leu Thr Leu Ala Phe Val Val Asn Ser Leu Leu Leu Ile 275 280 285 Leu Gly Ala Ala Leu Phe Phe Gly Thr Asn Ser Asp Leu Gly Arg Phe 290 295 300 Val Asp Leu Phe Asn Ala Leu Ser Asp Ser Gln Ile Val Gly Ala Ile 305 310 315 320 Ala Ser Pro Met Leu Ser Met Leu Phe Ala Leu Ala Leu Leu Ser Ser 325 330 335 Gly Gln Ser Ser Thr Ile Thr Gly Thr Leu Ala Gly Gln Ile Ile Met 340 345 350 Glu Gly Phe Ile Asn Leu Lys Met Pro Leu Trp Ala Gln Arg Leu Ile 355 360 365 Thr Arg Leu Leu Ser Val Thr Pro Val Ile Ile Phe Ala Ile Ile Tyr 370 375 380 His Gly Asn Glu Ala Lys Ile Glu Asp Leu Leu Thr Phe Ser Gln Val 385 390 395 400 Phe Leu Ser Ile Ala Leu Pro Phe Ala Met Ile Pro Leu Val Ile Phe 405 410 415 Thr Ser Ser Lys Lys Leu Met Gly Glu Phe Ala Asn Arg Thr Trp Ser 420 425 430 Lys Ile Leu Gly Trp Ile Ile Ala Val Ile Leu Ile Ile Leu Asn Ile 435 440 445 Tyr Leu Ile Leu Asn Thr Leu His Ile Val Gln 450 455 <210> 6 <211> 447 <212> PRT <213> Pediococcus acidilactici <400> 6 Met Ser Lys Lys Leu Asp Glu Val Asp Asn Lys Ser Leu Asp Glu Ile 1 5 10 15 Asn Gly Ser Ile Lys Val Pro Lys Asn Ala Gly Phe Phe Lys Thr Leu 20 25 30 Met Ala Tyr Thr Gly Pro Gly Ile Leu Ile Ala Val Gly Tyr Met Asp 35 40 45 Pro Gly Asn Trp Ile Thr Ser Ile Ala Gly Gly Ala Gln Phe Lys Tyr 50 55 60 Thr Leu Leu Ser Val Val Leu Ile Ser Ser Leu Ile Ala Met Leu Leu 65 70 75 80 Gln Ala Met Ser Ala Arg Leu Gly Ile Val Thr Gly Lys Asp Leu Ala 85 90 95 Gln Leu Thr Arg Glu Arg Thr Ser Lys Arg Val Gly Phe Met Leu Trp 100 105 110 Val Val Ala Glu Leu Ala Ile Met Ala Thr Asp Ile Ala Glu Ile Ile 115 120 125 Gly Ser Gly Ile Ala Leu Glu Leu Leu Phe His Ile Pro Leu Ile Ile 130 135 140 Gly Ile Leu Ile Thr Ala Ala Asp Val Leu Ile Leu Leu Leu Leu Met 145 150 155 160 Arg Leu Gly Phe Arg Lys Ile Glu Ala Ile Val Ala Thr Leu Val Met 165 170 175 Val Ile Leu Ile Val Phe Ala Tyr Glu Val Phe Leu Ser Asp Pro Ser 180 185 190 Ile Ser Gly Ile Ile Lys Gly Tyr Val Pro Ala Pro Val Ile Leu Gln 195 200 205 Asn Asn Ser Met Leu Tyr Leu Ser Leu Gly Ile Val Gly Ala Thr Val 210 215 220 Met Pro His Asp Leu Tyr Leu Gly Ser Ser Ile Ser Gln Thr Arg Glu 225 230 235 240 Ile Asp Arg Arg Asp Arg Lys Asn Val Ala Gln Ala Ile Arg Phe Ser 245 250 255 Thr Ile Asp Ser Asn Met Gln Leu Phe Leu Ala Phe Ile Val Asn Ser 260 265 270 Leu Leu Leu Ile Leu Gly Ala Ala Leu Phe Tyr Gly Thr Asp Ser Ser 275 280 285 Leu Gly Arg Phe Val Asp Leu Phe Asn Ala Leu Ser Asp Asn Gln Ile 290 295 300 Val Gly Ala Ile Ala Ser Pro Met Leu Ser Met Leu Phe Ala Val Ala 305 310 315 320 Leu Leu Ala Ser Gly Gln Ser Ser Thr Ile Thr Gly Thr Leu Ser Gly 325 330 335 Gln Ile Ile Met Glu Gly Phe Ile Arg Leu Arg Val Pro Leu Trp Val 340 345 350 Gln Arg Leu Val Thr Arg Leu Leu Ser Val Ala Pro Val Leu Ile Phe 355 360 365 Ala Ile Tyr Tyr His Gly Asp Glu Ala Lys Ile Glu Asn Leu Leu Thr 370 375 380 Phe Ser Gln Val Phe Leu Ser Val Ala Leu Pro Phe Ala Val Ile Pro 385 390 395 400 Leu Val Met Tyr Thr Ser Ser Lys Lys Leu Met Gly Glu Phe Ala Asn 405 410 415 Arg Gln Trp Val Lys Trp Cys Ala Trp Ile Ala Thr Ile Ile Leu Ile 420 425 430 Leu Leu Asn Ile Tyr Leu Ile Leu Gln Thr Leu Gly Ile Val Lys 435 440 445 <210> 7 <211> 464 <212> PRT <213> Lactobacillus plantarum <400> 7 Met Lys Ser Ala Lys Thr Lys Asp His Ala Lys Met Lys Ala Ala Glu 1 5 10 15 Glu Lys Ala Ile His Ser Thr Gly Ala Asp Ser Lys Ser Leu Asp Glu 20 25 30 Val Asn Gly Ser Val Arg Val Pro Lys Asp Ala Ser Phe Trp Arg Thr 35 40 45 Leu Ile Ala Tyr Thr Gly Pro Gly Ala Leu Val Ala Val Gly Tyr Met 50 55 60 Asp Pro Gly Asn Trp Ile Thr Ser Ile Ala Gly Gly Ser Gln Tyr Lys 65 70 75 80 Tyr Ala Leu Leu Ser Val Ile Leu Leu Ser Ser Leu Ile Ala Met Leu 85 90 95 Leu Gln Ala Met Ala Ala Arg Leu Gly Ile Val Thr Gly Lys Asp Leu 100 105 110 Ala Gln Leu Thr Arg Glu Arg Thr Ser Lys Gly Met Gly Ile Phe Leu 115 120 125 Trp Ile Ile Thr Glu Leu Ala Ile Met Ala Thr Asp Val Ala Glu Ile 130 135 140 Ile Gly Ser Gly Ile Ala Leu Lys Leu Leu Phe Gly Phe Pro Leu Ile 145 150 155 160 Val Gly Ile Leu Ile Thr Thr Ala Asp Val Leu Ile Leu Leu Leu Leu 165 170 175 Met Lys Leu Gly Phe Arg Lys Ile Glu Ala Ile Val Ala Thr Leu Val 180 185 190 Ala Val Ile Leu Phe Val Phe Leu Tyr Glu Val Ile Ile Ser Gln Pro 195 200 205 Asn Ile Pro Glu Met Leu Lys Gly Tyr Val Pro Thr Ser Arg Ile Val 210 215 220 Ser Asn Arg Ser Met Leu Phe Leu Ala Leu Gly Ile Val Gly Ala Thr 225 230 235 240 Val Met Pro His Asn Leu Tyr Leu Gly Ser Ser Ile Ser Gln Thr Arg 245 250 255 Gln Val Asp Arg Ser Asp Glu Lys Glu Val Ala Lys Ala Val Lys Phe 260 265 270 Thr Thr Ile Asp Ser Asn Ile Gln Leu Ser Val Ala Phe Val Val Asn 275 280 285 Ser Leu Leu Leu Ile Leu Gly Ala Ala Leu Phe Phe Gly Thr Lys Gly 290 295 300 Asp Leu Gly Arg Phe Val Asp Leu Tyr Asn Ala Leu Gly Asp Ser Lys 305 310 315 320 Val Val Gly Ser Ile Ala Ser Pro Leu Leu Ser Met Leu Phe Ala Ile 325 330 335 Ala Leu Leu Ser Ser Gly Gln Ser Ser Thr Ile Thr Gly Thr Leu Ser 340 345 350 Gly Gln Ile Ile Met Glu Gly Phe Ile Arg Leu Lys Met Pro Leu Trp 355 360 365 Ala Gln Arg Leu Leu Thr Arg Leu Ile Ser Val Thr Pro Val Leu Ala 370 375 380 Phe Ala Ile Tyr Tyr His Gly Asn Glu Ala Lys Ile Glu Asp Leu Leu 385 390 395 400 Thr Met Ser Gln Val Phe Leu Ser Ile Ala Leu Pro Phe Ala Met Ile 405 410 415 Pro Leu Val Met Phe Thr Ser Asn Arg Ala Leu Met Gly Asn Phe Thr 420 425 430 Asn Arg Val Trp Val Lys Trp Thr Ala Trp Ile Val Thr Val Ile Leu 435 440 445 Ile Ile Leu Asn Ile Tyr Leu Ile Leu Gln Thr Val Gly Leu Val Lys 450 455 460 <210> 8 <211> 443 <212> PRT <213> Lactobacillus sakei <400> 8 Met His Tyr Ala Asp Gly Ser Ser Leu Glu Glu Ile Asn Asn Thr Val 1 5 10 15 Ala Ile Pro Lys Asn Ala Gly Phe Trp Lys Thr Leu Met Ala Phe Met 20 25 30 Gly Pro Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp 35 40 45 Ile Thr Ser Ile Ala Gly Gly Ala Gln Phe Ala Tyr Thr Leu Ile Ser 50 55 60 Val Ile Leu Val Ser Asn Leu Ile Ala Met Leu Leu Gln Ala Met Ala 65 70 75 80 Ala Arg Leu Gly Ile Val Thr Gly Met Asp Leu Ala Gln Met Thr Arg 85 90 95 Ala Lys Thr Gly Lys Lys Met Gly Ile Phe Leu Trp Ile Val Thr Glu 100 105 110 Leu Ala Ile Met Ala Thr Asp Ile Ala Glu Ile Ile Gly Ser Ala Ile 115 120 125 Ala Leu Glu Leu Ile Phe Asn Ile Pro Leu Leu Trp Gly Val Leu Ile 130 135 140 Thr Ala Phe Asp Val Leu Leu Leu Leu Leu Leu Met Lys Leu Gly Phe 145 150 155 160 Arg Lys Ile Glu Ala Ile Val Ala Thr Leu Val Ala Val Ile Leu Phe 165 170 175 Val Phe Leu Tyr Glu Val Ile Leu Ala Gln Pro Asn Met Gly Asp Val 180 185 190 Val Arg Gly Phe Val Pro Ser Pro Arg Ile Met Thr Asp Lys Lys Met 195 200 205 Leu Phe Leu Ala Leu Gly Ile Val Gly Ala Thr Val Met Pro His Asn 210 215 220 Leu Tyr Leu His Ser Ser Ile Ala Gln Ala Arg Gln Tyr Asp Arg Asp 225 230 235 240 Asp Val Ala Glu Lys Arg Lys Ala Ile Lys Phe Thr Val Ile Asp Ser 245 250 255 Asn Ile Gln Leu Thr Ile Ala Phe Val Val Asn Cys Leu Leu Leu Ile 260 265 270 Leu Gly Ala Ala Met Phe Tyr Gly Thr Asn Ser Asp Leu Gly Arg Phe 275 280 285 Val Asp Leu Phe Asn Ala Leu Gln Asn Lys Glu Ile Val Gly Ser Ile 290 295 300 Ala Ser Pro Met Leu Ser Leu Leu Phe Ala Val Ala Leu Leu Ala Ser 305 310 315 320 Gly Gln Asn Ser Thr Ile Thr Gly Thr Leu Ser Gly Gln Ile Val Met 325 330 335 Glu Gly Phe Val Arg Met Lys Ile Pro Leu Trp Ala Arg Arg Val Ile 340 345 350 Thr Arg Gly Leu Ser Ile Leu Pro Val Ile Ile Phe Thr Val Tyr Tyr 355 360 365 His Gly Asn Glu Ala Gln Val Glu Asn Leu Leu Ile Tyr Ser Gln Val 370 375 380 Phe Leu Ser Ile Ala Leu Pro Val Ser Met Ile Pro Leu Thr Leu Phe 385 390 395 400 Thr Ser Asp Glu Lys Ile Met Gly Pro Phe Val Asn Arg Pro Trp Val 405 410 415 Lys Tyr Thr Ala Trp Phe Val Thr Ile Val Leu Thr Leu Leu Asn Ile 420 425 430 Tyr Leu Ile Leu Gln Thr Val Gly Leu Ala Ala 435 440 <210> 9 <211> 454 <212> PRT <213> Lactobacillus alimentarius <400> 9 Met Ser Ser Lys Asn Lys Lys His Glu Ser Leu Ile His Tyr Ala Asn 1 5 10 15 Gly Pro Ser Leu Glu Glu Ile Asn Asp Thr Val Glu Ile Pro Lys Asp 20 25 30 Ala Gly Phe Phe Lys Thr Leu Leu Ala Tyr Ser Gly Pro Gly Ala Leu 35 40 45 Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Val Thr Ser Ile Ala 50 55 60 Gly Gly Ala Gln Phe Lys Tyr Lys Leu Leu Ser Val Ile Leu Ile Ser 65 70 75 80 Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ser Ala Lys Leu Gly Ile 85 90 95 Val Thr Gly Arg Asp Leu Ala Gln Leu Thr Arg Asp Arg Thr Ser Arg 100 105 110 Val Gly Gly Phe Ile Leu Trp Ile Ile Thr Glu Leu Ala Ile Met Ala 115 120 125 Thr Asp Ile Ala Glu Ile Ile Gly Ser Ala Ile Ala Leu Lys Leu Leu 130 135 140 Phe Asn Ile Pro Val Leu Trp Gly Val Ile Ile Thr Ala Phe Asp Val 145 150 155 160 Leu Leu Leu Leu Val Leu Met Lys Leu Gly Phe Arg Lys Ile Glu Ala 165 170 175 Ile Val Ala Thr Leu Ile Met Val Ile Leu Leu Val Phe Leu Tyr Glu 180 185 190 Val Ile Leu Ala Lys Pro Asp Val Gly Gln Met Met Val Gly Phe Ile 195 200 205 Pro Glu Pro Lys Ile Leu Gln Asn Gln Ser Met Leu Tyr Leu Ser Leu 210 215 220 Gly Ile Val Gly Ala Thr Val Met Pro His Asn Leu Tyr Leu His Ser 225 230 235 240 Ser Ile Ser Gln Ala Arg Lys Tyr Asp Arg Asp Asp Pro Lys Ser Ile 245 250 255 His Gln Ala Val Arg Phe Ser Thr Trp Asp Ser Asn Ile Gln Leu Thr 260 265 270 Leu Ala Phe Val Val Asn Thr Leu Leu Leu Leu Leu Gly Ala Ala Leu 275 280 285 Phe Tyr Gly Thr Ser Ser Asp Leu Gly Arg Phe Val Asp Leu Phe Asn 290 295 300 Ala Leu Gln Asp Pro Lys Val Ala Gly Ala Val Ala Ser Pro Val Leu 305 310 315 320 Ser Ile Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr 325 330 335 Ile Thr Gly Thr Leu Ser Gly Gln Ile Val Met Glu Gly Phe Ile His 340 345 350 Met Lys Met Lys Leu Trp Ala Arg Arg Val Ile Thr Arg Leu Met Ser 355 360 365 Ile Ile Pro Val Ile Thr Phe Ala Ile Ile Tyr His Gly Asn Glu Ala 370 375 380 Lys Ile Glu Ser Leu Leu Thr Phe Ser Gln Val Phe Leu Ser Val Ala 385 390 395 400 Leu Pro Phe Ser Ile Phe Pro Leu Ile Lys Phe Thr Ser Asn Lys Lys 405 410 415 Leu Met Gly Glu Phe Val Asn Asn Lys Leu Val Glu Tyr Ile Gly Tyr 420 425 430 Phe Val Ala Ile Val Leu Thr Ile Leu Asn Ile Trp Leu Ile Tyr Thr 435 440 445 Thr Phe Val Pro Thr Ala 450 <210> 10 <211> 451 <212> PRT <213> Lactobacillus floricola <400> 10 Met Thr Lys Glu Glu Thr Lys Leu Phe His Tyr Ala Asp Gly Pro Ser 1 5 10 15 Leu Glu Glu Ile Asn Gly Thr Val Ala Val Pro Lys Lys Gly Gly Phe 20 25 30 Trp Lys Thr Leu Phe Ala Phe Ser Gly Pro Gly Ala Leu Val Ala Val 35 40 45 Gly Tyr Met Asp Pro Gly Asn Trp Val Thr Ser Ile Ala Gly Gly Ala 50 55 60 Gln Tyr Gln Tyr Thr Leu Leu Ser Val Ile Leu Ile Ser Ser Leu Ile 65 70 75 80 Ala Met Leu Leu Gln Ala Met Ser Ala Arg Leu Gly Ile Ala Ser Gly 85 90 95 Leu Asp Leu Ala Gln Ala Thr Ala Lys His Ser Pro Lys Trp Leu Arg 100 105 110 Tyr Thr Leu Trp Ile Ile Thr Glu Leu Ala Ile Met Ala Thr Asp Ile 115 120 125 Ala Glu Ile Val Gly Ala Ala Ile Ala Leu Lys Leu Leu Phe Asn Leu 130 135 140 Pro Leu Ile Val Gly Ile Phe Leu Thr Thr Leu Asp Val Met Leu Leu 145 150 155 160 Leu Leu Leu Met Lys Leu Gly Phe Arg Lys Ile Glu Ala Ile Val Gly 165 170 175 Ala Leu Ile Val Ser Ile Leu Val Ile Phe Leu Tyr Glu Val Ile Leu 180 185 190 Ala Arg Pro Asp Val Gly Ala Met Phe Ala Gly Tyr Ile Pro Gln Pro 195 200 205 Glu Val Val Thr Asn Lys Gly Ala Phe Tyr Ile Ala Leu Gly Ile Val 210 215 220 Gly Ala Thr Val Met Pro His Asn Leu Tyr Leu His Ser Ser Ile Ala 225 230 235 240 Gln Ala Arg Gln Tyr Asp Arg Asn Asp Ile Glu Glu Lys Lys Arg Ala 245 250 255 Ile Lys Phe Thr Val Leu Asp Ser Asn Ile Gln Leu Ser Val Ala Phe 260 265 270 Val Val Asn Thr Leu Leu Leu Leu Leu Gly Ala Ala Leu Phe Tyr Gly 275 280 285 Ala Gln Thr Asp Leu Gly Thr Phe Ser Glu Leu Tyr Asn Ala Leu Gln 290 295 300 Asn Pro Gln Val Ala Gly Val Ile Ala Ser Pro Ile Leu Ser Val Leu 305 310 315 320 Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile Thr Gly 325 330 335 Thr Leu Ser Gly Gln Ile Val Met Glu Gly Phe Ile His Leu Lys Met 340 345 350 Pro Met Trp Ala Arg Arg Val Ile Thr Arg Leu Ile Ser Val Ile Pro 355 360 365 Val Leu Ile Phe Ala Ile Ile Tyr His Ser Asn Glu Ala Lys Ile Glu 370 375 380 Asp Leu Leu Val Phe Ser Gln Val Phe Leu Ser Ile Ala Leu Pro Val 385 390 395 400 Ser Ile Ile Pro Leu Val Met Phe Thr Ala Asn Lys Lys Ile Met Gly 405 410 415 Pro Phe Val Asn Lys Lys Trp Val Thr Ile Thr Ser Ser Leu Val Ala 420 425 430 Ile Ile Leu Thr Gly Leu Asn Ile Phe Leu Ile Leu Gln Thr Leu Gly 435 440 445 Trp Val Gln 450 <210> 11 <211> 455 <212> PRT <213> Lactobacillus brevis <400> 11 Met Thr Asp Asn Val Ser Ala Lys Ser Val Gln Gly Asp Leu Thr Asn 1 5 10 15 Gly Pro Ser Leu Ala Glu Ile Asn Gly Ser Val Arg Val Pro Lys Glu 20 25 30 Lys Gly Phe Val Arg Asn Leu Leu Ala Phe Ser Gly Pro Gly Ala Leu 35 40 45 Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Val Thr Ser Ile Gly 50 55 60 Gly Gly Ala Gln Tyr Gly Tyr Leu Leu Met Ser Val Ile Leu Met Ser 65 70 75 80 Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys Leu Gly Ile 85 90 95 Val Thr Gln Met Asp Leu Ala Arg Ala Thr Arg Ala His Thr Gly Lys 100 105 110 Arg Ile Gly Ala Val Leu Trp Val Met Thr Glu Leu Ala Ile Met Ala 115 120 125 Thr Asp Ile Ala Glu Val Ile Gly Gly Ala Ile Ala Leu Lys Leu Leu 130 135 140 Phe Gly Val Pro Leu Ile Leu Gly Val Ser Leu Thr Val Leu Asp Val 145 150 155 160 Leu Leu Leu Leu Leu Leu Thr Arg Leu Gly Phe Arg Lys Ile Glu Ala 165 170 175 Ile Val Leu Cys Leu Ile Leu Val Ile Leu Val Val Phe Ala Tyr Glu 180 185 190 Val Val Ile Ala Gln Pro Ser Met Gly Gln Ala Val Ala Ser Phe Val 195 200 205 Pro Gln Ala Glu Ile Met Arg Pro Gly Gln Leu Thr Met Ala Leu Gly 210 215 220 Ile Val Gly Ala Thr Val Met Pro His Asn Leu Tyr Leu His Ser Ser 225 230 235 240 Ile Ala Gln Thr Arg Lys Phe Asp Arg Gln Asp Pro Ala Glu Met Ala 245 250 255 Arg Ala Val Lys Phe Thr Ala Trp Asp Ser Asn Ile Gln Leu Phe Gly 260 265 270 Ala Phe Ile Ile Asn Cys Leu Leu Leu Leu Leu Gly Ala Ala Met Phe 275 280 285 Phe Gly Lys Asp Ala Gly Ala Leu Gly Thr Phe Gly Gln Leu Tyr Asp 290 295 300 Ala Leu Gln Asp Asn Arg Leu Ala Gly Ala Val Ala Ser Pro Val Leu 305 310 315 320 Ser Thr Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr 325 330 335 Ile Thr Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Ile Asn 340 345 350 Met Arg Leu Pro Ile Trp Val Arg Arg Leu Val Thr Arg Leu Ile Ser 355 360 365 Val Ala Pro Val Ile Ile Val Thr Ile Leu Tyr Gly Gly Ser Glu Gln 370 375 380 Ala Leu Asp Arg Leu Leu Val Asn Ser Gln Val Phe Leu Ser Ile Ala 385 390 395 400 Leu Pro Phe Ser Met Ile Pro Leu Thr Ile Phe Thr Ser Ser Lys Arg 405 410 415 Ile Met Gly Thr Arg Trp Val Asn Arg Trp Trp Val Thr Ala Leu Ala 420 425 430 Trp Gly Cys Thr Ala Ile Leu Thr Val Leu Asn Ile Gln Ile Val Trp 435 440 445 Ala Thr Met Thr Thr Leu Phe 450 455 <210> 12 <211> 450 <212> PRT <213> Lactobacillus casei <400> 12 Met Ala Arg Pro Asp Glu Arg Leu Thr Val Gln Arg Glu Lys Arg Ser 1 5 10 15 Leu Asp Asp Ile Asn Arg Ser Val Gln Val Pro Ser Val Tyr Glu Ser 20 25 30 Ser Phe Phe Gln Lys Phe Leu Ala Tyr Ser Gly Pro Gly Ala Leu Val 35 40 45 Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Leu Thr Ala Leu Glu Gly 50 55 60 Gly Ser Arg Tyr His Tyr Ala Leu Leu Ser Val Leu Leu Met Ser Ile 65 70 75 80 Leu Val Ala Met Phe Met Gln Thr Leu Ala Ile Lys Leu Gly Val Val 85 90 95 Ala Arg Leu Asp Leu Ala Gln Ala Ile Ala Ala Phe Ile Pro His Trp 100 105 110 Ser Arg Ile Cys Leu Trp Leu Ile Asn Glu Ala Ala Met Met Ala Thr 115 120 125 Asp Met Thr Gly Val Val Gly Thr Ala Ile Ala Leu Lys Leu Leu Phe 130 135 140 Gly Leu Pro Leu Met Trp Gly Met Leu Leu Thr Ile Ala Asp Val Leu 145 150 155 160 Val Val Leu Leu Phe Leu Arg Phe Gly Ile Arg Arg Val Glu Leu Ile 165 170 175 Val Leu Val Ser Ile Leu Thr Val Gly Ile Ile Phe Gly Ile Glu Val 180 185 190 Ala Arg Ala Asp Pro Ser Ile Gly Gly Ile Ala Gly Gly Phe Val Pro 195 200 205 His Thr Asp Ile Leu Thr Asn His Gly Met Leu Leu Leu Ser Leu Gly 210 215 220 Ile Met Gly Ala Thr Ile Met Pro His Asn Ile Tyr Leu His Ser Ser 225 230 235 240 Leu Ala Gln Ser Arg Lys Tyr Asp Glu His Ile Pro Ala Gln Val Thr 245 250 255 Glu Ala Leu Arg Phe Gly Lys Trp Asp Ser Asn Val His Leu Val Ala 260 265 270 Ala Phe Leu Ile Asn Ala Leu Leu Leu Ile Leu Gly Ala Ala Leu Phe 275 280 285 Tyr Gly Val Gly Gly His Val Thr Ala Phe Gln Gly Val Tyr Asn Gly 290 295 300 Leu Lys Asn Pro Met Ile Val Gly Gly Leu Ala Ser Pro Leu Met Ser 305 310 315 320 Thr Leu Phe Ala Phe Ala Leu Leu Ile Thr Gly Leu Ile Ser Ser Ile 325 330 335 Ala Ser Thr Leu Ala Gly Gln Ile Val Met Glu Gly Tyr Leu Asn Ile 340 345 350 Arg Met Pro Leu Trp Glu Arg Arg Leu Leu Thr Arg Leu Val Thr Leu 355 360 365 Ile Pro Ile Met Val Ile Gly Phe Met Ile Gly Phe Ser Glu His Asn 370 375 380 Phe Glu Gln Val Ile Val Tyr Ala Gln Val Ser Leu Ser Ile Ala Leu 385 390 395 400 Pro Phe Thr Leu Phe Pro Leu Val Ala Leu Thr Asn Arg Arg Asp Leu 405 410 415 Met Gly Ile His Val Asn Ser Gln Leu Val Arg Trp Val Gly Tyr Phe 420 425 430 Leu Thr Gly Val Ile Thr Val Leu Asn Ile Gln Leu Ala Ile Ser Val 435 440 445 Phe Val 450 <210> 13 <211> 451 <212> PRT <213> Lactobacillus rhamnosus <400> 13 Met Thr Lys Arg Asn Glu Gln Leu Ser Val Gln Gln Ala Lys Pro Ser 1 5 10 15 Leu Asp Glu Ile Asn Arg Ser Val Gln Val Pro Gly Val Tyr Glu Pro 20 25 30 Ser Phe Val Gln Lys Phe Leu Ala Tyr Ser Gly Pro Gly Ala Leu Val 35 40 45 Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Leu Thr Ala Leu Glu Gly 50 55 60 Gly Ser Arg Tyr His Glu Thr Leu Leu Ala Val Leu Leu Leu Ser Ile 65 70 75 80 Leu Ala Ala Met Phe Met Gln Thr Leu Ala Ile Lys Leu Gly Val Val 85 90 95 Ala Arg Leu Asp Leu Ala Gln Ala Ile Ala Ala Phe Val Pro Lys Trp 100 105 110 Ser Arg Ile Gly Leu Trp Leu Val Asn Glu Ala Ala Met Met Ala Thr 115 120 125 Asp Met Thr Gly Val Val Gly Thr Ala Ile Ala Leu Lys Leu Leu Phe 130 135 140 Gly Leu Pro Leu Met Trp Gly Met Leu Leu Thr Ile Ala Asp Val Leu 145 150 155 160 Val Val Leu Met Phe Leu Arg Phe Gly Ile Arg Arg Ile Glu Leu Ile 165 170 175 Val Leu Ala Ser Ile Leu Thr Val Gly Ile Ile Phe Gly Ile Glu Val 180 185 190 Val Arg Ala Arg Pro Ser Met Gly Gly Ile Val Ala Gly Leu Val Pro 195 200 205 His Thr Glu Ile Leu Thr Asn Arg Gly Met Leu Leu Leu Ser Leu Gly 210 215 220 Ile Met Gly Ala Thr Ile Met Pro His Asn Ile Tyr Leu His Ser Ser 225 230 235 240 Leu Ala Gln Ser Arg Arg Tyr Asp Glu His Ile Pro Ala Gln Val Thr 245 250 255 Glu Ala Leu Arg Phe Gly Lys Trp Asp Ser Asn Val His Leu Val Ala 260 265 270 Ala Phe Ile Ile Asn Ala Leu Leu Leu Ile Leu Gly Ala Thr Leu Phe 275 280 285 Tyr Gly Met Ser Ser His Ala Thr Ala Phe Glu Gly Val Tyr Asn Gly 290 295 300 Leu Lys Asn Pro Ala Ile Val Gly Gly Leu Ala Ser Pro Leu Met Ser 305 310 315 320 Thr Leu Phe Ala Phe Ala Leu Leu Ile Thr Gly Leu Ile Ser Ser Ile 325 330 335 Ala Ser Thr Leu Ala Gly Gln Ile Val Met Glu Gly Tyr Leu Asn Ile 340 345 350 Gln Ile Pro Leu Trp Ala Arg Arg Leu Leu Thr Arg Leu Val Thr Leu 355 360 365 Ile Pro Ile Met Ile Ile Gly Phe Val Met Gly Phe Ser Glu Gln His 370 375 380 Phe Glu Gln Val Ile Val Tyr Ala Gln Val Ala Leu Ser Ile Ala Leu 385 390 395 400 Pro Phe Thr Leu Phe Pro Leu Val Ala Leu Thr Asp Arg Arg Asp Leu 405 410 415 Met Gly Gln His Val Asn Ser Pro Val Val Arg Trp Met Gly Tyr Val 420 425 430 Leu Thr Gly Ile Ile Thr Leu Leu Asn Val Gln Leu Ile Leu Ser Val 435 440 445 Ile Leu Pro 450 <210> 14 <211> 442 <212> PRT <213> Lactobacillus kefiri <400> 14 Met Ser Gln Glu Pro Thr His Lys Ser Leu Asp Glu Ile Asn Gln Ser 1 5 10 15 Val Glu Val Pro Ser Val Tyr Glu Thr Ser Phe Leu Gln Lys Phe Leu 20 25 30 Ala Tyr Ser Gly Pro Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro 35 40 45 Gly Asn Trp Leu Thr Ser Leu Ser Gly Gly Ser Gln Phe Arg Tyr Ala 50 55 60 Leu Leu Ser Val Leu Leu Met Ser Ile Leu Val Ala Met Phe Met Gln 65 70 75 80 Thr Leu Ser Ile Lys Leu Gly Val Val Ala Arg Leu Asp Leu Ala Gln 85 90 95 Ala Ile Ala Gln Lys Val Pro Lys Ser Gly Arg Tyr Thr Leu Trp Ile 100 105 110 Ile Asn Glu Leu Ala Met Met Ala Thr Asp Met Thr Gly Val Val Gly 115 120 125 Thr Ala Ile Ala Leu Lys Leu Leu Phe Gly Leu Pro Leu Val Tyr Gly 130 135 140 Ile Leu Leu Thr Ile Phe Asp Val Leu Leu Val Leu Leu Phe Leu Arg 145 150 155 160 Phe Gly Ile Arg Arg Ile Glu Phe Ile Val Leu Ala Ala Ile Leu Ile 165 170 175 Val Gly Val Ile Phe Gly Ile Glu Val Thr Arg Ala Thr Pro Asn Ile 180 185 190 Val Glu Ile Ala Gly Gly Leu Ile Pro Thr Thr His Ile Val Thr Asn 195 200 205 His Glu Met Leu Ile Met Ser Leu Gly Ile Val Gly Ala Thr Ile Met 210 215 220 Pro His Asn Val Tyr Leu His Ser Ser Leu Ala Gln Ser Arg Arg Tyr 225 230 235 240 Asp Tyr His Asn Pro Lys Gln Val Asn Glu Ala Leu Arg Phe Ala Lys 245 250 255 Trp Asp Ser Asn Val His Leu Val Ala Ala Phe Leu Ile Asn Ala Leu 260 265 270 Leu Leu Val Leu Gly Gly Thr Leu Phe Phe His Thr Asn Ser His Phe 275 280 285 Ser Ala Phe Gln Asp Val Tyr Asn Gly Leu Lys Ser Ser Ala Ile Val 290 295 300 Gly Ser Leu Ala Ser Pro Leu Met Ser Thr Leu Phe Ala Phe Ala Leu 305 310 315 320 Leu Ile Thr Gly Met Ile Ser Ser Ile Thr Ser Thr Leu Ser Gly Gln 325 330 335 Ile Val Met Glu Gly Tyr Leu His Ile Arg Leu Pro Leu Trp Glu Arg 340 345 350 Arg Leu Leu Thr Arg Phe Val Thr Leu Ile Pro Ile Leu Ala Ile Gly 355 360 365 Phe Leu Val Gly Phe Asn Asp His Asp Phe Glu Glu Ile Ile Val Tyr 370 375 380 Ala Gln Ile Ala Leu Ser Ile Ala Leu Pro Phe Thr Leu Phe Pro Met 385 390 395 400 Val Ala Leu Thr Ser Asn His Asp Leu Met Gly Val His Thr Asn Arg 405 410 415 Arg Tyr Val Thr Val Ile Gly Tyr Leu Leu Thr Ser Ile Ile Thr Ile 420 425 430 Leu Asn Leu Gln Phe Val Leu Ala Ser Ile 435 440 <210> 15 <211> 439 <212> PRT <213> Lactobacillus digestum <400> 15 Met Pro Asn Lys Lys Ser Leu Asp Glu Ile Asn Glu Ser Val Lys Val 1 5 10 15 Pro Ser Val Tyr Asp Thr Ser Phe Leu Gln Lys Phe Leu Ala Tyr Ser 20 25 30 Gly Pro Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp 35 40 45 Leu Thr Ser Leu Ser Gly Gly Ser Gln Tyr Arg Tyr Asp Leu Leu Ser 50 55 60 Val Leu Leu Ile Ser Ile Leu Val Ala Met Phe Met Gln Thr Leu Ser 65 70 75 80 Ile Lys Leu Gly Val Val Ala Arg Leu Asp Leu Ala Gln Ala Ile Ala 85 90 95 Thr Lys Val Ser Lys Pro Ile Arg Tyr Phe Leu Trp Ile Leu Asn Glu 100 105 110 Ile Ala Met Met Ala Thr Asp Leu Thr Gly Val Ile Gly Thr Ala Ile 115 120 125 Ala Leu Lys Leu Leu Phe Asn Leu Pro Leu Val Phe Gly Ile Leu Leu 130 135 140 Thr Val Phe Asp Val Leu Ile Val Leu Ile Phe Leu Arg Phe Gly Ile 145 150 155 160 Arg Arg Ile Glu Phe Ile Val Leu Ala Ala Ile Leu Thr Val Gly Ile 165 170 175 Ile Phe Gly Ile Glu Val Phe Arg Ala Gln Pro Lys Leu Phe Ser Ile 180 185 190 Ile Ser Gly Val Ile Pro Ser Thr Asp Leu Phe Thr Asn His Arg Lys 195 200 205 Leu Val Leu Ser Leu Gly Ile Val Gly Ala Thr Ile Met Pro His Asn 210 215 220 Ile Tyr Leu His Ser Ser Leu Ala Gln Ser Arg Arg Tyr Asp His Asn 225 230 235 240 Asp Pro Leu Gln Val Asn Glu Ala Leu Arg Phe Ala Lys Trp Asp Ser 245 250 255 Asn Val His Leu Ile Ala Ala Phe Ile Ile Asn Ala Leu Leu Leu Val 260 265 270 Leu Gly Gly Thr Leu Phe Tyr His Met Thr Asn Gln Leu Ala Ser Leu 275 280 285 Gln Asp Val Phe Thr Gly Leu Lys Ser His Ala Ile Val Gly Thr Leu 290 295 300 Ala Ser Pro Leu Met Ser Trp Leu Phe Ala Phe Ala Leu Leu Ile Thr 305 310 315 320 Gly Met Ile Ser Ser Ile Thr Ser Thr Leu Ser Gly Gln Ile Val Met 325 330 335 Glu Gly Tyr Leu Asn Ile Arg Leu Pro Leu Trp Gln Arg Arg Leu Leu 340 345 350 Thr Arg Phe Val Thr Leu Ile Pro Ile Leu Ile Ile Gly Phe Ile Val 355 360 365 His Phe Asn Glu Gln Asp Phe Glu Asn Leu Ile Val Tyr Ala Gln Ile 370 375 380 Ile Leu Ser Ile Ala Leu Pro Phe Thr Leu Phe Pro Met Ile Phe Leu 385 390 395 400 Thr Asn Asp Lys Lys Ile Met Gly Asn His Val Asn Ser Lys Leu Thr 405 410 415 Thr Thr Val Gly Ile Ile Leu Ala Ser Ala Ile Thr Ile Leu Asn Leu 420 425 430 Gln Leu Leu Phe Ser Leu Ile 435 <210> 16 <211> 441 <212> PRT <213> Lactobacillus plantarum <400> 16 Met Gln Ser His Arg His Gln Ser Leu Glu Glu Ile Asn Gln Ser Val 1 5 10 15 Ala Val Pro Asp Val His Gln Thr Ala Phe Trp Arg Lys Phe Leu Ala 20 25 30 Tyr Ser Gly Pro Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly 35 40 45 Asn Trp Leu Thr Ser Leu Ala Gly Gly Gly Gln Phe Gln Tyr Arg Leu 50 55 60 Leu Ala Val Leu Ala Leu Ala Ile Ile Val Ala Met Phe Met Gln Gly 65 70 75 80 Leu Ala Ile Arg Leu Gly Val Val Ala Arg Gln Asp Leu Ala Gln Ala 85 90 95 Ile Ala Ser Lys Leu Pro Arg Pro Val Arg Tyr Ala Ala Trp Ile Leu 100 105 110 Asn Glu Val Ala Met Met Ala Thr Asp Met Thr Gly Val Ile Gly Thr 115 120 125 Ala Ile Ala Leu Lys Met Leu Phe Gly Leu Pro Leu Leu Ala Gly Ile 130 135 140 Leu Leu Thr Ile Ala Asp Val Leu Val Val Leu Leu Phe Leu Arg Phe 145 150 155 160 Gly Ile Arg Arg Val Glu Val Ile Val Leu Val Ala Ile Leu Thr Val 165 170 175 Gly Ile Ile Phe Gly Ile Glu Val Gly Arg Ala His Val Gln Phe Gly 180 185 190 Asn Val Leu Leu Gly Leu Val Pro Thr Pro Leu Ile Val Lys Asn His 195 200 205 Thr Ala Leu Val Leu Ser Leu Gly Ile Leu Gly Ala Thr Ile Met Pro 210 215 220 His Asn Leu Tyr Leu His Ser Ser Leu Ala Gln Ser Arg Arg Tyr Asp 225 230 235 240 Tyr His Asn Pro Ala Gln Val Thr Glu Ala Leu Arg Phe Ala Asn Trp 245 250 255 Asp Ser Thr Val His Leu Ile Ala Ala Phe Leu Ile Asn Ala Leu Leu 260 265 270 Leu Val Leu Gly Gly Thr Leu Phe Phe Gly His Thr Asn Ala Leu Ala 275 280 285 Ser Leu Gln Ala Val Phe Asp Gly Leu Lys Ser Thr Thr Val Val Gly 290 295 300 Ala Leu Ala Ser Pro Val Met Ser Trp Leu Phe Ala Leu Ala Leu Leu 305 310 315 320 Ile Thr Gly Leu Ile Ser Ser Ile Thr Ser Thr Leu Ala Gly Gln Ile 325 330 335 Val Met Glu Gly Tyr Leu His Ile Arg Leu Pro Leu Trp Gln Arg Arg 340 345 350 Leu Leu Thr Arg Ala Val Thr Leu Ile Pro Ile Leu Ile Ile Gly Met 355 360 365 Leu Val Gly Phe Ser Asp Ala Ala Phe Glu Asn Leu Ile Ile Tyr Ala 370 375 380 Gln Val Ala Leu Ser Ile Ala Leu Pro Phe Thr Leu Leu Pro Leu Val 385 390 395 400 Ala Leu Thr Asn Asp Ala Ser Leu Met Lys Ala His Val Asn Arg Pro 405 410 415 Ala Val Thr Trp Val Gly Tyr Gly Leu Ala Gly Ile Ile Thr Val Leu 420 425 430 Asn Ile Tyr Leu Val Tyr Ser Leu Phe 435 440 <210> 17 <211> 434 <212> PRT <213> Lactobacillus reuteri <400> 17 Met Glu Arg Lys Ser Leu Asp Glu Ile Asn Gly Ser Val Asp Val Pro 1 5 10 15 Asn Val Tyr Gln Ser Ala Phe Trp Gln Lys Phe Leu Ala Tyr Ser Gly 20 25 30 Pro Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Leu 35 40 45 Thr Ser Leu Ala Gly Gly Ser Gln Tyr Arg Tyr Gln Leu Leu Val Val 50 55 60 Leu Phe Thr Ala Ile Leu Ile Ala Met Tyr Met Gln Ser Leu Ala Ile 65 70 75 80 Lys Leu Gly Val Thr Thr Arg Thr Asp Leu Ala Gln Ala Ile Ala Arg 85 90 95 Arg Leu Pro Thr Pro Leu Arg Ile Ala Leu Trp Leu Phe Asn Glu Ile 100 105 110 Ala Met Met Ala Thr Asp Leu Thr Gly Val Val Gly Thr Ala Val Ala 115 120 125 Leu Asn Met Leu Phe Lys Leu Pro Leu Leu Ile Gly Val Leu Leu Thr 130 135 140 Ile Ala Asp Val Leu Val Val Leu Phe Phe Leu His Phe Gly Ile Arg 145 150 155 160 Arg Ile Glu Phe Ile Val Leu Thr Ala Ile Leu Val Val Gly Ala Ile 165 170 175 Phe Ala Ile Glu Val Cys Arg Ala His Pro Glu Phe Ser Ala Ile Met 180 185 190 Asp Gly Phe Val Pro Arg Ser Thr Ile Phe Thr Asn His Ser Glu Leu 195 200 205 Leu Ile Ser Leu Gly Ile Val Gly Ala Thr Ile Met Pro His Asn Ile 210 215 220 Tyr Leu His Ser Ser Leu Ala Gln Ser Arg Arg Tyr Asp Glu His Asp 225 230 235 240 Pro Lys Gln Val Lys Glu Thr Leu Arg Phe Ala Asn Trp Asp Ser Leu 245 250 255 Ile His Leu Phe Ala Ala Phe Ile Val Asn Ala Leu Leu Leu Ile Leu 260 265 270 Gly Gly Thr Leu Phe Phe His Ala Ala Ser Leu Gly Ser Leu Glu Asp 275 280 285 Val Phe Phe Gly Leu Lys Asn Pro Gln Ile Val Gly Ser Leu Ala Ser 290 295 300 Pro Leu Met Ser Trp Leu Phe Ala Phe Ala Leu Leu Val Thr Gly Leu 305 310 315 320 Ile Ser Ser Ile Thr Ser Thr Leu Ala Gly Gln Ile Val Met Glu Gly 325 330 335 Phe Ile Asn Ile Arg Leu Pro Leu Trp Lys Arg Arg Leu Leu Thr Arg 340 345 350 Ala Val Thr Leu Val Pro Ile Leu Ile Ile Gly Phe Met Ile Asn Phe 355 360 365 Lys Glu Glu Gln Phe Glu Gln Leu Ile Ile Tyr Ala Gln Ile Val Leu 370 375 380 Ser Ile Ala Leu Pro Phe Thr Leu Tyr Pro Leu Val Ala Leu Thr Gly 385 390 395 400 Asn Lys Lys Leu Met Gly Pro His Val Asn Ser Arg Trp Gln Thr Val 405 410 415 Leu Gly Tyr Ile Leu Ala Ser Leu Val Thr Gly Leu Asn Leu Leu Val 420 425 430 Leu Val <210> 18 <211> 440 <212> PRT <213> Lactobacillus crustorum <400> 18 Met Thr Glu Lys Lys Ser Leu Asp Glu Ile Asn Gly Ser Val Ala Val 1 5 10 15 Pro Gln Tyr Asn Thr Ser Phe Phe Arg Lys Phe Leu Ala Tyr Ser Gly 20 25 30 Pro Gly Ala Leu Ile Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Leu 35 40 45 Thr Ser Leu Val Gly Gly Ala His His Lys Tyr Gln Leu Leu Ser Val 50 55 60 Leu Leu Ile Ser Ile Leu Val Ala Thr Phe Met Gln Ser Leu Ser Ile 65 70 75 80 Arg Leu Gly Ile Ala Ser Arg Gln Asp Leu Ala Gln Ala Ile Ala Lys 85 90 95 Lys Ala Lys Lys Pro Val Arg Tyr Cys Leu Trp Ile Ile Asn Glu Leu 100 105 110 Ala Met Met Ala Thr Asp Leu Thr Gly Val Ile Gly Thr Ala Leu Ala 115 120 125 Leu Asn Met Leu Phe Lys Leu Pro Leu Val Phe Gly Val Leu Ile Thr 130 135 140 Ile Leu Asp Val Phe Leu Ile Leu Trp Phe Met Arg Phe Gly Ile Arg 145 150 155 160 Arg Ile Glu Ser Ile Val Val Ile Ser Ile Leu Thr Val Gly Leu Ile 165 170 175 Phe Ala Phe Glu Val Ser His Val Gln Pro Asn Leu Thr Ala Ile Phe 180 185 190 Lys Gly Phe Val Pro Ser Gln Thr Ile Ile Thr Asn Gln Asn Lys Leu 195 200 205 Ile Leu Ser Leu Gly Ile Ile Gly Ala Thr Ile Met Pro His Asn Ile 210 215 220 Tyr Leu His Ser Ala Leu Ala Gln Ser Arg Arg Tyr Asp Tyr His Asp 225 230 235 240 Ser Arg Gln Val Arg Glu Ala Leu Arg Phe Ala Asn Trp Asp Ser Ile 245 250 255 Val His Leu Ile Ala Ala Leu Ile Ile Asn Cys Leu Leu Leu Ile Leu 260 265 270 Gly Gly Thr Ile Phe Tyr Asp Lys Ala Asp Gln Leu Ala Ser Leu Met 275 280 285 Thr Val Phe Lys Gly Leu Met Asn Tyr Gln Val Val Gly Ser Leu Ala 290 295 300 Ser Ser Phe Met Ser Tyr Leu Phe Ala Phe Ala Leu Leu Val Thr Gly 305 310 315 320 Leu Ile Ser Ser Ile Thr Ser Thr Leu Ser Gly Gln Ile Val Met Glu 325 330 335 Gly Tyr Leu Asn Ile Arg Leu Pro Leu Trp Gln Arg Arg Leu Leu Thr 340 345 350 Arg Ile Ile Thr Leu Ile Pro Ile Leu Val Ile Gly Phe Leu Val His 355 360 365 Phe Asn Glu Val Ile Phe Glu Asp Leu Ile Val Tyr Ala Gln Ile Ala 370 375 380 Leu Ser Val Ala Leu Pro Phe Thr Leu Phe Pro Leu Val Tyr Leu Thr 385 390 395 400 Asn Asn Ala Lys Ile Met Gly Lys His Val Asn Lys Lys Trp Gln Thr 405 410 415 Ile Leu Gly Phe Val Leu Ala Leu Ile Ile Thr Ile Leu Asn Ile Val 420 425 430 Leu Ile Ala Thr Thr Leu Ser His 435 440 <210> 19 <211> 535 <212> PRT <213> Lactobacillus casei <400> 19 Met Ser Asp Asp His Lys Lys Arg His Pro Ile Lys Leu Ile Gln Tyr 1 5 10 15 Ala Asn Gly Pro Ser Leu Glu Glu Ile Asn Gly Thr Val Glu Val Pro 20 25 30 His Gly Lys Gly Phe Trp Arg Thr Leu Phe Ala Tyr Ser Gly Pro Gly 35 40 45 Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr Ser 50 55 60 Ile Thr Gly Gly Gln Asn Phe Gln Tyr Leu Leu Ile Ser Val Ile Leu 65 70 75 80 Met Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys Leu 85 90 95 Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg Thr 100 105 110 Ser Lys Lys Leu Gly Ile Val Leu Trp Ile Leu Thr Glu Leu Ala Ile 115 120 125 Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu Tyr 130 135 140 Leu Leu Phe His Ile Pro Leu Val Ile Ala Val Leu Val Thr Val Leu 145 150 155 160 Asp Val Leu Val Leu Leu Leu Leu Thr Lys Ile Gly Phe Arg Lys Ile 165 170 175 Glu Ala Ile Val Val Ala Leu Ile Leu Val Ile Leu Leu Val Phe Val 180 185 190 Tyr Gln Val Ala Leu Ser Asp Pro Asn Met Gly Ala Leu Leu Lys Gly 195 200 205 Phe Ile Pro Thr Gly Glu Thr Phe Ala Ser Ser Pro Ser Ile Asn Gly 210 215 220 Met Ser Pro Ile Gln Gly Ala Leu Gly Ile Ile Gly Ala Thr Val Met 225 230 235 240 Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln Thr Arg Lys Ile 245 250 255 Asp His Lys Asn Pro Asp Asp Val Ala Gln Ala Val Lys Phe Ser Ala 260 265 270 Trp Asp Ser Asn Ile Gln Leu Ser Phe Ala Phe Val Val Asn Cys Leu 275 280 285 Leu Leu Val Met Gly Val Ala Val Phe Lys Ser Gly Ala Val Lys Asp 290 295 300 Pro Ser Phe Phe Gly Leu Phe Gln Ala Leu Ser Asp Ser Ser Thr Leu 305 310 315 320 Ser Asn Gly Val Leu Ile Ala Val Ala Lys Ser Gly Ile Leu Ser Ile 325 330 335 Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile Thr 340 345 350 Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Val His Met Lys 355 360 365 Methionine, Proline, Leucine, Tryptophan, Alanine, Arginine, Arginine, Leucine, Valine, Threonine, Arginine, Isoleucine, Isoleucine, Serine, Valine, Isoleucine 370 375 380 Proline, Valine, Isoleucine, Valine, Cysteine, Valine, Methionine, Leucine, Threonine, Alanine, Arginine, Aspartic acid, Threonine, Proline, Isoleucine, Glutamine 385 390 395 400 Glutamine, Histidine, Glutamic acid, Alanine, Leucine, Asparagine, Threonine, Leucine, Methionine, Asparagine, Asparagine, Serine, Glutamine, Valine, Phenylalanine, Leucine 405 410 415 Alanine, Phenylalanine, Alanine, Leucine, Proline, Phenylalanine, Serine, Methionine, Leucine, Proline, Leucine, Leucine, Methionine, Phenylalanine, Threonine, Asparagine 420 425 430 Serine, Lysine, Valine, Glutamic acid, Methionine, Glycine, Aspartic acid, Arginine, Phenylalanine, Lysine, Asparagine, Threonine, Glycine, Tryptophan, Valine, Lysine 435 440 445 Valine, Leucine, Glycine, Tryptophan, Isoleucine, Serine, Valine, Leucine, Glycine, Leucine, Threonine, Glycine, Leucine, Asparagine, Leucine, Lysine 450 455 460 Glycine, Leucine, Proline, Aspartic acid, Serine, Isoleucine, Alanine, Glycine, Phenylalanine, Phenylalanine, Glycine, Aspartic acid, Histidine, Proline, Threonine, Alanine 465 470 475 480 Threonine, Glutamine, Threonine, Asparagine, Methionine, Alanine, Asparagine, Isoleucine, Isoleucine, Alanine, Isoleucine, Valine, Leucine, Isoleucine, Valine, Alanine 485 490 495 Isoleucine, Leucine, Alanine, Leucine, Leucine, Alanine, Tryptophan, Threonine, Isoleucine, Tryptophan, Aspartic acid, Leucine, Tyrosine, Lysine, Glycine, Asparagine 500 505 510 Glutamine, Arginine, Tyrosine, Glutamic acid, Alanine, Histidine, Leucine, Alanine, Alanine, Valine, Alanine, Aspartic acid, Glutamic acid, Lysine, Glutamic acid, Alanine 515 520 525 Lys Ala Asp Val Asp Glu Gln 530 535 <210> 20 <211> 535 <212> PRT <213> Lactobacillus rhamnosus <400> 20 Met Ser Asp Asp His Lys Lys Lys His Ser Met Lys Leu Ile Gln Tyr 1 5 10 15 Ala Asn Gly Pro Ser Leu Glu Glu Ile Asn Gly Thr Val Glu Val Pro 20 25 30 His Gly Lys Gly Phe Trp Arg Thr Leu Phe Ala Tyr Ser Gly Pro Gly 35 40 45 Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr Ser 50 55 60 Ile Thr Gly Gly Gln Asn Phe Gln Tyr Leu Leu Ile Ser Val Ile Leu 65 70 75 80 Met Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys Leu 85 90 95 Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg Thr 100 105 110 Ser Lys Lys Leu Gly Ile Val Leu Trp Ile Leu Thr Glu Leu Ala Ile 115 120 125 Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu Tyr 130 135 140 Leu Leu Phe His Ile Pro Leu Val Ile Ala Val Leu Val Thr Val Leu 145 150 155 160 Asp Val Leu Val Leu Leu Leu Leu Thr Lys Ile Gly Phe Arg Lys Ile 165 170 175 Glu Ala Ile Val Val Ala Leu Ile Leu Val Ile Leu Leu Val Phe Val 180 185 190 Tyr Gln Val Ala Leu Ser Asp Pro Asn Met Gly Ala Leu Leu Lys Gly 195 200 205 Phe Ile Pro Thr Gly Glu Thr Phe Ala Ser Ser Pro Ser Val Asn Gly 210 215 220 Met Ser Pro Ile Gln Gly Ala Leu Gly Ile Ile Gly Ala Thr Val Met 225 230 235 240 Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln Thr Arg Lys Ile 245 250 255 Asp His Lys Asp Pro Glu Asp Val Ala Gln Ala Val Lys Phe Ser Ala 260 265 270 Trp Asp Ser Asn Ile Gln Leu Thr Phe Ala Phe Val Val Asn Cys Leu 275 280 285 Leu Leu Val Met Gly Val Ala Val Phe Lys Ser Gly Ala Val Lys Asp 290 295 300 Pro Ser Phe Phe Gly Leu Phe Gln Ala Leu Ser Asp Ser Ser Thr Leu 305 310 315 320 Ser Asn Gly Val Leu Ile Ala Val Ala Lys Ser Gly Ile Leu Ser Ile 325 330 335 Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile Thr 340 345 350 Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Ile His Met Lys 355 360 365 Met Pro Leu Trp Ala Arg Arg Leu Val Thr Arg Val Ile Ser Val Ile 370 375 380 Pro Val Ile Val Cys Val Met Leu Thr Ala Arg Glu Thr Pro Ile Gln 385 390 395 400 Gln His Glu Ala Leu Asn Thr Leu Met Asn Asn Ser Gln Val Phe Leu 405 410 415 Ala Phe Ala Leu Pro Phe Ser Met Leu Pro Leu Leu Met Phe Thr Asn 420 425 430 Ser Lys Val Glu Met Gly Asp Arg Phe Lys Asn Thr Gly Trp Val Lys 435 440 445 Val Leu Gly Trp Val Ser Val Ile Gly Leu Thr Tyr Leu Asn Leu Lys 450 455 460 Gly Leu Pro Asp Ser Ile Ala Gly Phe Phe Gly Asp Asn Pro Thr Ala 465 470 475 480 Ala Gln Thr Asn Ile Ala Asn Met Ile Ala Tyr Val Leu Ile Ala Ala 485 490 495 Val Leu Ala Leu Leu Ala Trp Thr Ile Trp Asp Leu Tyr Lys Gly Asn 500 505 510 Lys Arg Tyr Glu Ala His Leu Glu Ala Val Ala Asp Glu Glu Glu Ala 515 520 525 Lys Ala Asn Asp Asp Val Gln 530 535 <210> 21 <211> 530 <212> PRT <213> Lactobacillus plantarum <400> 21 Met Ser Glu Lys Thr Asn Thr Pro Asn Arg Lys His Lys Leu Ile Glu 1 5 10 15 Tyr Ala Asn Gly Pro Ser Leu Glu Glu Ile Asn Gly Thr Ile Glu Val 20 25 30 Pro Lys Asn Leu Asn Phe Trp Lys Thr Leu Phe Ala Tyr Ser Gly Pro 35 40 45 Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr 50 55 60 Ser Ile Thr Gly Gly Gln Asn Tyr Gln Tyr Met Leu Met Ser Val Ile 65 70 75 80 Leu Ile Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys 85 90 95 Leu Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg 100 105 110 Thr Ser Lys Ser Leu Gly Ile Val Leu Trp Ile Leu Thr Glu Leu Ala 115 120 125 Ile Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu 130 135 140 Tyr Leu Leu Phe Asn Ile Pro Leu Val Ile Ala Val Phe Ile Thr Val 145 150 155 160 Leu Asp Val Leu Val Leu Leu Leu Leu Thr Lys Ile Gly Phe Arg Lys 165 170 175 Ile Glu Ala Ile Val Val Cys Leu Ile Leu Val Ile Leu Phe Val Phe 180 185 190 Val Tyr Gln Val Ala Leu Ser Asn Pro Asp Trp Gly Gly Val Ile Lys 195 200 205 Gly Leu Val Pro Thr Ala Asp Thr Phe Ser Thr Ser Arg Ser Val Asn 210 215 220 Gly Met Thr Pro Leu Ser Gly Ala Leu Gly Ile Ile Gly Ala Thr Val 225 230 235 240 Met Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln Thr Arg Lys 245 250 255 Ile Asp His Asn Asp Glu Glu Asp Val Ala Arg Thr Val Lys Phe Ala 260 265 270 Ala Trp Asp Ser Asn Ile Gln Leu Ser Phe Ala Phe Val Val Asn Ser 275 280 285 Leu Leu Leu Ile Met Gly Val Ala Val Phe Lys Ser Gly Ala Val Lys 290 295 300 Asp Pro Ser Phe Phe Gly Leu Tyr Glu Ala Leu Ser Asn Thr Ser Met 305 310 315 320 Leu Ser Asn Gly Ile Leu Ile Ser Val Ala Lys Ser Gly Ala Leu Ser 325 330 335 Ala Leu Phe Ala Ile Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile 340 345 350 Thr Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Val His Met 355 360 365 Arg Met Pro Leu Trp Leu Arg Arg Leu Val Thr Arg Leu Ile Ser Val 370 375 380 Ile Pro Val Leu Ile Cys Val Leu Leu Thr Ser Gly Lys Ser Ala Ile 385 390 395 400 Asp Glu His Thr Ala Leu Asn Asn Leu Met Asn Asn Ser Gln Val Phe 405 410 415 Leu Ala Phe Ala Leu Pro Phe Ser Met Leu Pro Leu Leu Met Met Thr 420 425 430 Asp Ser Ala Ala Glu Met Gly Lys Arg Phe Lys Asn Ser Leu Trp Ile 435 440 445 Lys Gly Leu Gly Trp Leu Ser Val Ile Gly Leu Thr Phe Leu Asn Leu 450 455 460 Leu Gly Leu Pro Asp Ser Ile Leu Gly Phe Phe Gly Asp Asn Pro Ser 465 470 475 480 Ala Gly Glu Gln Thr Phe Ser Lys Ile Leu Ala Tyr Leu Leu Ile Ala 485 490 495 Ala Ile Leu Ala Leu Leu Val Trp Thr Val Phe Asp Leu Gln Arg Gly 500 505 510 Asn Lys Arg Tyr Val Glu Gln Gln Leu Ala Ala Ala Ala Lys Glu Ala 515 520 525 Asn Lys 530 <210> 22 <211> 530 <212> PRT <213> Pediococcus acidilactici <400> 22 Met Ser Asn Glu Ile Lys Asn Pro Lys Lys Arg Arg Lys Leu Ile Ser 1 5 10 15 Tyr Ala Asn Gly Arg Ser Leu Glu Glu Ile Asn Gly Thr Val Lys Val 20 25 30 Pro Lys Asn Ile Gly Phe Trp Lys Thr Leu Phe Met Tyr Ser Gly Pro 35 40 45 Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr 50 55 60 Ser Ile Thr Gly Gly Gln Asn Phe Gln Tyr Met Leu Met Ser Ile Ile 65 70 75 80 Leu Ile Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys 85 90 95 Leu Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg 100 105 110 Thr Ser Arg Ala Leu Gly Ile Val Leu Trp Ile Leu Thr Glu Leu Ala 115 120 125 Ile Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu 130 135 140 Tyr Leu Leu Phe His Ile Pro Leu Val Val Ala Val Phe Ile Thr Val 145 150 155 160 Phe Asp Val Leu Leu Leu Leu Leu Leu Thr Lys Ile Gly Phe Arg Lys 165 170 175 Ile Glu Ala Ile Val Val Cys Leu Ile Met Val Ile Leu Val Val Phe 180 185 190 Val Tyr Gln Val Ala Leu Ser His Pro Ser Trp Gly Ala Val Phe Gly 195 200 205 Gly Leu Ile Pro Thr Thr Lys Ala Phe Ala Thr Thr Pro Thr Val Gly 210 215 220 Gly Met Thr Pro Leu Ser Gly Ser Leu Gly Ile Ile Gly Ala Thr Val 225 230 235 240 Met Pro His Asn Leu Tyr Leu His Ser Ala Val Ser Gln Thr Arg Lys 245 250 255 Ile Asn His Asp Asp Glu Glu Asp Val Ala Arg Thr Val Arg Phe Ser 260 265 270 Thr Trp Asp Ser Asn Ile Gln Leu Ser Phe Ala Phe Val Val Asn Ala 275 280 285 Leu Leu Leu Val Met Gly Val Ala Val Phe Lys Thr Gly Ala Val Gln 290 295 300 Asp Pro Ser Phe Phe Gly Leu Phe His Ala Leu Asn Asp Thr Ser Thr 305 310 315 320 Leu Ser Asn Gly Ile Leu Ile Gly Val Ala Lys Thr Gly Ile Leu Ser 325 330 335 Thr Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile 340 345 350 Thr Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Val His Met 355 360 365 Arg Met Pro Leu Trp Ala Arg Arg Leu Ile Thr Arg Leu Ile Ser Val 370 375 380 Val Pro Val Leu Ile Cys Val Met Leu Thr Ser Gly Lys Gly Thr Ile 385 390 395 400 Gln Glu His Glu Ala Leu Asn Asn Leu Met Asn Asn Ser Gln Val Phe 405 410 415 Leu Ala Phe Ala Leu Pro Phe Ser Met Val Pro Leu Leu Met Met Thr 420 425 430 Asp Ser Arg Val Glu Met Gly Asp Arg Phe Lys Asn Ser Trp Ile Val 435 440 445 Arg Ile Leu Gly Trp Ile Ser Val Ile Phe Leu Thr Tyr Leu Asn Leu 450 455 460 Thr Gly Leu Pro Asp Ser Ile Ala Ala Phe Phe Gly Glu Asn Ala Ser 465 470 475 480 Ala Ala Glu Ile Ser Met Ala His Asp Ile Ala Tyr Ala Leu Ile Val 485 490 495 Ala Val Leu Ala Leu Leu Ala Trp Thr Val Ile Glu Leu Tyr Lys Gly 500 505 510 Asn Lys Arg Tyr Glu Ile Glu Leu Ala Glu Lys Ala Asn Ala Lys Glu 515 520 525 Ala Ala 530 <210> 23 <211> 521 <212> PRT <213> Lactobacillus salivarius <400> 23 Met Val Asn Asn Glu Asn Asn His Lys Lys His Lys Met Ile Gln Tyr 1 5 10 15 Ala Asn Gly Lys Ser Leu Glu Glu Ala Asn Gly Thr Val Glu Ile Pro 20 25 30 Lys Gly Lys Gly Phe Trp Lys Thr Leu Phe Ala Tyr Ser Gly Pro Gly 35 40 45 Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr Ser 50 55 60 Ile Thr Gly Gly Gln Asn Phe Gln Tyr Leu Leu Met Ser Val Ile Leu 65 70 75 80 Leu Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys Leu 85 90 95 Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg Thr 100 105 110 Ser Lys Ala Leu Gly Ile Val Leu Trp Ile Leu Thr Glu Leu Ala Ile 115 120 125 Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu Tyr 130 135 140 Leu Leu Phe Asp Ile Pro Leu Ile Ile Ala Val Phe Ile Thr Val Phe 145 150 155 160 Asp Val Leu Leu Leu Leu Leu Leu Thr Lys Val Gly Phe Arg Lys Ile 165 170 175 Glu Ala Ile Val Val Cys Leu Ile Phe Val Ile Leu Phe Val Phe Val 180 185 190 Tyr Gln Val Ala Leu Ser Asn Pro Asp Trp Gly Gly Val Phe Lys Gly 195 200 205 Leu Ile Pro Thr Ser Glu Thr Phe Ala Lys His Pro Val Val His Asp 210 215 220 Met Ser Pro Leu Asn Gly Ala Leu Gly Ile Ile Gly Ala Thr Val Met 225 230 235 240 Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln Thr Arg Lys Phe 245 250 255 Asp Arg Asn Asn Glu Asp Asp Ile Ala Asn Ala Val Arg Phe Thr Ala 260 265 270 Trp Asp Ser Asn Ile Gln Leu Gly Leu Ala Phe Val Val Asn Ser Leu 275 280 285 Leu Leu Ile Met Gly Val Ala Val Phe Lys Ser Gly Ala Val Glu Asp 290 295 300 Pro Ser Phe Phe Gly Leu Tyr Gln Ala Leu Ser Asp Thr Ser Val Met 305 310 315 320 Ser Asn Gly Leu Leu Ala Ala Ala Ala Arg Thr Gly Ile Leu Ser Thr 325 330 335 Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile Thr 340 345 350 Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Ile His Leu Arg 355 360 365 Methionine, Proline, Leucine, Tryptophan, Alanine, Arginine, Arginine, Leucine, Isoleucine, Threonine, Arginine, Leucine, Leucine, Serine, Valine, Isoleucine 370 375 380 Proline, Valine, Leucine, Isoleucine, Cysteine, Valine, Alanine, Leucine, Threonine, Serine, Glycine, Lysine, Serine, Threonine, Isoleucine, Glutamic acid 385 390 395 400 Glutamic acid, Histidine, Glutamic acid, Alanine, Leucine, Asparagine, Asparagine, Leucine, Methionine, Asparagine, Asparagine, Serine, Glutamine, Valine, Phenylalanine, Leucine 405 410 415 Alanine, Phenylalanine, Alanine, Leucine, Proline, Phenylalanine, Serine, Methionine, Leucine, Proline, Leucine, Valine, Isoleucine, Methionine, Threonine, Glycine 420 425 430 Serine, Lysine, Valine, Glutamic acid, Methionine, Glycine, Glutamic acid, Arginine, Phenylalanine, Lysine, Asparagine, Arginine, Leucine, Tryptophan, Isoleucine, Asparagine 435 440 445 Isoleucine, Leucine, Glycine, Tryptophan, Isoleucine, Serine, Valine, Isoleucine, Serine, Leucine, Threonine, Tyrosine, Leucine, Asparagine, Methionine, Isoleucine 450 455 460 Glycine, Leucine, Proline, Glutamine, Asparagine, Leucine, Glutamic acid, Proline, Phenylalanine, Phenylalanine, Proline, Alanine, Aspartic acid, Lysine, Valine, Glycine 465 470 475 480 Leucine, Alanine, Histidine, Threonine, Valine, Alanine, Tyrosine, Isoleucine, Leucine, Isoleucine, Valine, Leucine, Isoleucine, Isoleucine, Alanine, Leucine 485 490 495 Leucine, Isoleucine, Tryptophan, Threonine, Leucine, Valine, Glutamic acid, Leucine, Histidine, Leucine, Glycine, Asparagine, Lysine, Arginine, Phenylalanine, Alanine 500 505 510 Alanine, Glutamic acid, Glutamine, Alanine, Lysine, Lysine, Histidine, Asparagine, Lysine 515 520 <210> 24 <211> 520 <212> PRT <213> Lactobacillus fermentum <400> 24 Met Asp Asn Thr Lys Asn Gln His Arg Lys Leu Arg Leu Ile Glu His 1 5 10 15 Ala Asn Gly Lys Ser Leu Glu Glu Ile Asn Gly Thr Val Glu Val Pro 20 25 30 His Gly Lys Gly Phe Phe Arg Thr Leu Phe Ala Tyr Ser Gly Pro Gly 35 40 45 Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn Trp Ser Thr Ser 50 55 60 Ile Thr Gly Gly Gln Ser Phe Gln Tyr Thr Leu Met Thr Thr Ile Leu 65 70 75 80 Ile Ser Ser Leu Ile Ala Met Leu Leu Gln Tyr Met Ala Ala Lys Leu 85 90 95 Gly Ile Val Ser Gln Met Asp Leu Ala Gln Ala Ile Arg Ala Arg Thr 100 105 110 Gly Lys Ala Leu Gly Val Ile Leu Trp Leu Met Thr Glu Leu Ala Ile 115 120 125 Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala Ile Ala Leu Asn 130 135 140 Leu Leu Phe His Ile Pro Leu Val Leu Ala Val Phe Ile Thr Val Leu 145 150 155 160 Asp Val Leu Val Leu Leu Leu Leu Thr Lys Ile Gly Phe Arg Lys Ile 165 170 175 Glu Ala Ile Val Ala Cys Leu Ile Leu Val Ile Leu Ala Val Phe Ala 180 185 190 Tyr Gln Val Ala Leu Ser His Pro Asp Trp Ala Gly Val Phe Lys Gly 195 200 205 Leu Leu Pro Thr Lys Glu Ala Ile Ala Lys Glu Pro Val Val Gly Gly 210 215 220 Ile Ser Pro Leu Thr Gly Ser Leu Gly Ile Ile Gly Ala Thr Val Met 225 230 235 240 Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln Thr Arg Lys Ile 245 250 255 Asp His Thr Asn Ala Glu Asp Ile Lys Gln Thr Val Arg Phe Thr Ala 260 265 270 Trp Asp Ser Asn Ile Gln Leu Thr Leu Ala Phe Phe Val Asn Ala Leu 275 280 285 Leu Leu Ile Met Gly Val Ala Val Phe Lys Asn Gly Ala Val Gln Asp 290 295 300 Ser Ser Phe Phe Gly Leu Tyr Asp Ala Leu Asn Asn Thr Asp Met Leu 305 310 315 320 Ser Asn Gly Leu Leu Ile Ala Val Ala Lys Ser Gly Val Leu Ser Thr 325 330 335 Leu Phe Ala Ile Ala Leu Leu Ala Ser Gly Gln Asn Ser Thr Ile Thr 340 345 350 Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Phe Val His Met Lys 355 360 365 Met Pro Leu Trp Ala Arg Arg Leu Ile Thr Arg Leu Leu Ser Val Val 370 375 380 Pro Val Leu Val Cys Val Ala Met Thr Ala His Glu Ser Thr Ile Asp 385 390 395 400 Gln His Ala Ser Leu Asn Ile Leu Met Glu Asn Ser Gln Val Phe Leu 405 410 415 Ala Phe Ala Leu Pro Phe Ser Met Leu Pro Leu Leu Ile Met Thr Asn 420 425 430 Ser Asp Thr Glu Met Gly Gln Phe Lys Asn Ser Leu Trp Val Arg Val 435 440 445 Leu Gly Trp Ile Ser Val Ile Gly Leu Thr Phe Leu Asn Leu Tyr Asn 450 455 460 Leu Pro Gln Thr Tyr Glu Gly Phe Gly Ile Trp Ser Lys Gly Leu Ser 465 470 475 480 Asp Val Leu Ala Trp Ile Ser Ile Val Val Ile Val Val Leu Leu Ala 485 490 495 Trp Thr Cys Phe Glu Leu Ile Arg Gly Asp Arg Arg Leu Ala Ala Glu 500 505 510 Arg Glu Lys His Thr Trp Glu Lys 515 520 <210> 25 <211> 533 <212> PRT <213> Lactobacillus amylolyticus <400> 25 Met Cys Ser Arg Lys Val Leu Leu Thr Lys Gln Lys Gly Lys His Tyr 1 5 10 15 Leu Ile Arg Tyr Ala Asn Gly Lys Ser Leu Ser Glu Ile Asn Gly Thr 20 25 30 Ile Glu Ile Pro Lys Lys Arg Thr Phe Trp Arg Met Leu Trp Ala Tyr 35 40 45 Thr Gly Pro Gly Ala Leu Val Ala Val Gly Tyr Met Asp Pro Gly Asn 50 55 60 Trp Ala Thr Ser Ile Thr Gly Gly Gln Ser Phe Gln Tyr Ile Leu Met 65 70 75 80 Ser Thr Ile Leu Ile Ser Ser Leu Met Ala Met Leu Leu Gln Tyr Met 85 90 95 Ala Ala Lys Leu Gly Ile Val Thr Gln Met Asp Leu Ala Gln Ala Ile 100 105 110 Arg Leu Arg Thr Gly Lys Ala Leu Gly Ile Val Leu Trp Leu Met Thr 115 120 125 Glu Leu Ala Ile Met Ala Thr Asp Ile Ala Glu Val Ile Gly Ala Ala 130 135 140 Ile Ala Leu Asn Leu Leu Phe Asp Ile Pro Leu Val Pro Ala Val Phe 145 150 155 160 Ile Thr Val Leu Asp Val Leu Leu Leu Leu Leu Leu Ala Arg Ile Gly 165 170 175 Phe Arg Lys Ile Glu Ala Val Val Ser Cys Leu Ile Leu Val Ile Leu 180 185 190 Leu Val Phe Val Tyr Glu Val Leu Leu Ser Asn Pro Asp Trp Ser Lys 195 200 205 Ala Phe Val Gly Leu Val Pro Ser Ala Lys Ile Ile Gln Thr His Pro 210 215 220 Val Val Gly Gly Ile Ser Pro Leu Thr Gly Thr Leu Gly Ile Ile Gly 225 230 235 240 Ala Thr Val Met Pro His Asn Leu Tyr Leu His Ser Ala Ile Ser Gln 245 250 255 Thr Arg Lys Ile Asn His His Asn Leu Gln Leu Ile Arg Asp Ala Val 260 265 270 Lys Tyr Thr Ala Leu Asp Ser Asn Ile Gln Leu Ser Leu Ala Phe Leu 275 280 285 Val Asn Ala Leu Leu Leu Ile Met Gly Ala Ala Val Phe Lys Ser Gly 290 295 300 Ala Val Arg Asp Ser Ser Phe Phe Gly Leu Tyr Gln Ala Leu Asp Asn 305 310 315 320 Ala Lys Met Leu Ser Asp Pro Leu Leu Val His Val Ala Arg Thr Gly 325 330 335 Ile Leu Ser Thr Leu Phe Ala Val Ala Leu Leu Ala Ser Gly Gln Asn 340 345 350 Ser Thr Ile Thr Gly Thr Leu Thr Gly Gln Val Ile Met Glu Gly Tyr 355 360 365 Ile His Leu Lys Met Pro Leu Trp Ala Arg Arg Leu Val Thr Arg Leu 370 375 380 Leu Ser Val Ile Pro Val Leu Leu Cys Val Ser Phe Thr Met Asn Asp 385 390 395 400 Ser Val Met Gln Gln His Phe Ala Leu Asn Met Leu Met Glu Asn Ser 405 410 415 Gln Val Phe Leu Ala Phe Ala Leu Pro Phe Ser Val Leu Pro Leu Leu 420 425 430 Ile Met Thr Asn Asn Lys Ala Glu Met Gly Glu Phe Lys Asn Lys Pro 435 440 445 Leu Trp His Tyr Leu Gly Trp Ala Cys Ala Leu Val Leu Thr Phe Leu 450 455 460 Asn Leu Tyr Asn Leu Pro Ser Gln Phe Val Asn Phe Lys Phe Ala Ser 465 470 475 480 Lys Glu Val Ser Thr Ile Ile Ala Tyr Phe Val Ile Val Val Ile Ala 485 490 495 Ala Leu Leu Leu Trp Thr Cys Ile Glu Ile Tyr Ile Gly Asp Arg Lys 500 505 510 Val Lys Ile His His Ser Gly Phe Asp Ala Lys Glu Lys Glu Leu Lys 515 520 525 Glu Glu Gly Gln Lys 530

Claims

1. Use of one or more bacterial strains as manganese scavengers to inhibit or delay fungal growth in fermented dairy products, wherein the one or more bacterial strains are selected from Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus sakei, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus alimentarius, Pediococcus acidilactici, Lactobacillus rhamnosus, and Lactobacillus kefiri, wherein the one or more bacterial strains are added to the fermented dairy product to reduce the free manganese in the fermented dairy product to a concentration of less than 0.01 ppm, and wherein the fungus is yeast or mold.

2. Use of one or more bacterial strains as manganese scavengers to inhibit or delay the growth of fungi in fermented dairy products, wherein the one or more bacterial strains are selected from Lactobacillus fermentum, Lactobacillus reuteri, Lactobacillus sakei, Lactobacillus brevis, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus alimentarius, Pediococcus acidilactici, Lactobacillus rhamnosus, and Lactobacillus kefiri, wherein the one or more bacterial strains and a chemical chelating material are added to the fermented dairy product to reduce the free manganese in the fermented dairy product to a concentration below 0.01 ppm, wherein the fungi are yeasts or molds, and wherein the chemical chelating material is selected from ethylenediaminetetraacetic acid, ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid, diaminocyclohexanetetraacetic acid, nitrilotriacetic acid, 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid, and diethylenetriaminepentaacetic acid.

3. The use according to claim 2, wherein, The fungus is a yeast selected from the group consisting of species of Torulaspora, Cryptococcus, Saccharomyces, Yarrowia, Debaryomyces, Candida, and Rhodotorula, or wherein the fungus is a mold selected from the group consisting of species of Aspergillus, Cladosporium, Didymella, and Penicillium.

4. The use according to claim 1, wherein, The fermented dairy product is a thermophilic fermented food or a mesophilic fermented food, wherein the thermophilic fermented food is a product prepared by fermentation with a thermophilic organism, which is a microorganism that grows best at a temperature above 43 °C, and wherein the mesophilic fermented food is a product prepared by fermentation with a mesophilic organism, which is a microorganism that grows best at a moderate temperature of 15 °C - 40 °C.

5. The use according to claim 1, wherein the fermented dairy product has a pH of 3.5 to 6.

5.

6. The use according to claim 1, wherein the fermented dairy product has a water activity (aw) of less than 0.

98. w ) 7. The use according to claim 5 or 6, wherein the fermented dairy product is selected from the group consisting of quark cheese, cream cheese, fresh cheese, Greek yogurt, skimmed buttermilk, concentrated yogurt, buttermilk, sour cream, kefir, Indian yogurt, salted yogurt, tvorog, mint salted yogurt, smetana, Yakult, and dahi.

8. The use according to claim 5 or 6, wherein the fermented dairy product is yogurt.

9. The use according to claim 5 or 6, wherein the fermented dairy product is fermented milk.

10. Use according to claim 5 or 6, wherein the fermented dairy product is selected from the group consisting of European cheese, fresh cheese, soft cheese, cheddar cheese, mascarpone cheese, pastor cheese, mozzarella cheese, pizza cheese, feta cheese, brie cheese, camembert cheese, cottage cheese, red wave cheese, gouda cheese, thuringer cheese, havarti cheese or emmental cheese, Swiss cheese, and marscapone cheese.

11. Use according to claim 1 or 2, wherein, The free manganese in the fermented dairy product is reduced to a concentration of less than 0.005 ppm.

12. Use according to claim 2, wherein, The step of reducing free manganese in the fermented dairy product includes using ion exchange chromatography.

13. Use according to claim 2, wherein, The chemical chelating material is ethylenediaminetetraacetic acid.

14. Use according to claim 1 or 2, wherein, The one or more bacterial strains comprise a manganese transporter.

15. Use according to claim 1 or 2, wherein the one or more bacterial strains do not contain superoxide dismutase.

16. Use according to claim 1 or 2, wherein the one or more bacterial strains do not contain manganese superoxide dismutase.

17. Use according to claim 1 or 2, wherein, The manganese uptake activity of the one or more bacterial strains is measured.

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