Process for producing novel yeast and mold inhibiting products

Claim 2

-2- A process for producing yeast and mold inhibiting products (FIC) which comprises: (a) incubating live cells of a Lactobacillus species in a nutrient medium for the cells containing growth factors present in cysteine, garlic extract, milk, whey, yeast, yeast extract, molasses or protein digest which induce the formation of the products (FIC), a protein source and a carbon source, so as to produce the products (FIC) in the nutrient medium; and (b) treating the growth medium which has been incubated so as to produce the products (FIC) with or without the live cells, wherein the products (FIC) inhibit Penicillium oxalicum spores in an assay with the products (FIC) and the Penicillium oxalicum spores mixed.

Claim 3

-3- The process of Claim 2 wherein the nutrient medium which is incubated contains a yeast extract for the growth factors, and non-fat dry milk as a protein source; corn steep as a source of essential minerals and lactose, sucrose, dextrose, fructose or molasses as the carbon source.

Claim 4

-4- The process of Claim 3 wherein the nutrient medium contains a compound selected from the group consisting cinnamic acid, an alkali metal propionate and phenylalanine, in amounts which increase the production of the products (FIC).

Claim 5

-5- The process of Claim 2 wherein after the nutrient medium is incubated, the growth medium with the live cells is dried.

Claim 6

-6- The products (FIC) of the process of Claim 1.

Claim 7

-7- A process for producing yeast and mold inhibiting products (FIC) which comprises: (a) incubating live cells of a Lactobacillus having yeast and mold inhibiting characteristics of Lactobacillus casei var. rhamnosus NRRL-B-15972 in a nutrient medium for the cells containing growth factors present in cysteine, garlic extract, milk, whey, yeast, yeast extract, molasses or protein digest which induce the formation of the products (FIC), a protein source and a carbon source so as to produce products (FIC) in the nutrient medium wherein the products (FIC) inhibit Penicillium oxalicum spores in an assay with the products (FIC) and the Penicillium oxalicum spores mixed; and (b) treating the nutrient medium which has been incubated so as to produce the products (FIC) with the cells disrupted, wherein the products (FIC) inhibit Penicillium oxalicum spores in an assay with the products (FIC) and the Penicillium oxalicum spores mixed.

Claim 8

-8- The process of Claim 7 wherein after the nutrient medium is incubated, the nutrient medium with the live cells of Lactobacillus is dried.

Claim 9

-9- The process of Claim 7 wherein the Lactobacillus are incubated in the nutrient medium at a temperature of between about 10° and 50°C and then dried to produce a concentrate of the products (FIC).

Claim 10

-10- The process of Claim 7 wherein the nutrient medium contains yeast extract, whey or non-fat dry milk as a protein source, corn steep as a source of essential salts and lactose, sucrose, dextrose, fructose or molasses as the carbon source.

Claim 11

-11- The process of claim 10 wherein the nutrient medium contains a manganese salt and a magnesium salt as essential salts and an alkali metal phosphate as a buffer.

Claim 12

-12- The products (FIC) of the process of Claim 7.

Claim 13

-13- A process for producing yeast and mold inhibiting products (FIC) which comprises: (a) incubating cells of a Lactobacillus having yeast and mold inhibiting characteristics of Lactobacillus casei var. rhamnosus NRRL-B-15972 in a nutrient medium for the cells containing growth factors present in cysteine, garlic extract, milk, whey, yeast, yeast extract, molasses or protein digest which induce the formation of the products (FIC) and a carbon source so as to produce the products (FIC) in the nutrient medium wherein the products (FIC) inhibit Penicillium oxalicum spores in an assay with the products (FIC) and the Penicillium oxalicum spores mixed; (b) mixing the incubated nutrient medium with a water immiscible organic solvent which extracts the product into the solvent; (c) separating the solvent with the extracted product from the nutrient medium; and (d) separating the products (FIC) from the solvent.

Claim 14

-14- The process of Claim 13 wherein the solvent is n-butanol.

Claim 15

-15- The process of Claim 13 wherein the solvent is acetone.

Claim 16

-16- The process of Claim 13 wherein the solvent is ethanol.

Claim 17

-17- The process of Claim 13 wherein the solvent mixture is cooled to thereby precipitate impurities.

Claim 18

-18- The process of Claim 13 wherein in addition the separated products are purified to separate impurities using liquid chromatography.

Claim 19

-19- The products (FIC) produced by the process of Claim 13, wherein the products (FIC) have a primary ultraviolet absorbance at about 269 nanometers.

Claim 20

-20- The process of Claim 13 wherein in addition the products (FIC) are purified by liquid dialysis which passes compounds through a membrane having a molecular size of about 1000 or less and wherein the products pass through the membrane and are collected.

Claim 21

-21- The purified products (FIC) of the process of Claim 20.

Claim 22

-22- The process of Claim 20 wherein in addition the purified products (FIC) are further purified by liquid chromatography using a solvent mixture to produce separation of the products (FIC) from impurities.

Claim 23

-23- The process of Claim 22 wherein in addition the products (FIC) without the impurities are repurified using liquid chromatography.

Claim 24

-24- The process of Claim 13 wherein the nutrient medium which is incubated contains growth promoting amounts of a sulfur containing organic compound, yeast extract and protein digest.

Claim 25

-25- The process of Claim 1 wherein the nutrient medium which is to be incubated is pasteurized or sterilized at about 100°C or above prior to incubating the cells.

Claim 26

-26- A method for preventing yeast and mold growth in a material in need thereof which comprises: adding to the material products (FIC) produced by a process which comprises incubating live cells of a Lactobacillus in a nutrient medium containing a sulfur containing organic compound, a protein source and a carbon source, which induce the formation of the yeast and mold inhibiting products (FIC) in the nutrient medium wherein the products (FIC) in the nutrient medium inhibit Penicillium oxalicum spores in an assay with the products (FIC) and the Penicillium oxalicum spores mixed and then treating the nutrient medium which now contains live cells, in such a manner so as to produce the final products (FIC) in a form containing live cells, dead cells or no cells to thereby prevent yeast and mold growth in the material.

Claim 27

-27- The method of Claim 26 wherein the material is a food.

Claim 28

-28- The method of Claim 27 wherein the food is a carbonated beverage, Cottage cheese, yogurt, margarine, bread, grains and nuts.

Claim 29

-29- The method of Claim 26 wherein the material is living tissue.

Claim 30

-30- The method of Claim 27 wherein the material is a cleanser.

Claim 31

-31- A process for producing yeast and mold inhibiting products (FIC) which comprises: (a) incubating cells of a Lactobacillus having yeast and mold inhibiting characteristics of Lactobacillus casei var. rhamnosus NRRL-B-15972 in a nutrient medium for the cells containing growth factors present in cysteine, garlic extract, milk, whey, yeast, yeast extract, molasses or protein digest which induce the formation of the products (FIC), a protein source and a carbon source so as to produce the products (FIC) in the nutrient medium wherein the products (FIC) inhibit Penicillium oxalicum spores in an assay with the products (FIC) and the Penicillium oxalicum spores mixed; and (b) separating the products (FIC) from the nutrient medium and cells.

Claim 32

A process for producing yeast and mold inhibiting products (FIC) which comprises: (a) incubating live cells of a Lactobacillus species in a nutrient medium for the cells so as to produce an isolatable amount of the products (FIC) in the nutrient medium, wherein the products (FIC) produced in the medium or separated from the medium inhibit Penicillium oxalicum up to a level of about 10⁷ spores per ml when mixed in an agar-broth medium.

Claim 33

-33- A composition containing the products (FIC) produced by the process of Claim 32.