# US 10,370,636 B2 · Starter culture compositions > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,370,636 B2 - Document ID: US10370636B2 - Status: Granted - Kind: B2 - Application number: 13/990,742 - Filed: 2011-12-08 - Published: 2013-12-19 - Granted: 2019-08-06 - Earliest claimed: 2010-12-10 - Assignees: Dsm IP Assets B.V. - Applicants: Pim Van Hee - Inventors: Pim Van Hee - Web page: https://patentgrep.com/patent/US10370636B2 ## CPC - C12N 1/04: Preserving or maintaining viable microorganisms (immobilised microorganisms C12N11/00) - A23C 9/123: using only microorganisms of the genus lactobacteriaceae; Yoghurt (A23C9/13 takes precedence) - A23C 9/1238: {using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus} - C12N 1/38: Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound (C12N1/34 takes precedence) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A23C: DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; PREPARATION THEREOF (preservation thereof A23B11/00) ## Abstract The present invention relates to frozen and freeze dried starter culture compositions comprising microorganisms. The invention also relates to methods of producing the frozen and freeze dried starter culture compositions and methods of using them. ## Claims ### Claim 1 A process for making a starter culture composition comprising a microorganism, a cryoprotectant and at least one stimulating additive, said process comprising: a. culturing a microorganism in a culture medium; and b. collecting the microorganism from the culture medium; and c. adding a cryoprotectant to the microorganism obtained in b); and d. freezing said microorganism obtained in c) in the form of pellets or granules; and e. adding at least one frozen stimulating additive in the form of pellets or granules to a microorganism obtained in d), wherein no stimulating additive is added before step d), and wherein said freezing of step d) is performed directly following the addition of cryoprotectant of step c); wherein the ratio of stimulating additive (wt.) to microorganism (wt.) in the starter culture composition is between 0.1 and 10; and wherein the addition of at least one frozen stimulating additive of step e) increases the acidification rate of the starter culture composition compared to addition of the stimulating additive before freezing, and wherein the cryoprotectant is added in an amount sufficient to increase the acidification rate of the starter culture composition compared to a composition without the added cryoprotectant. ### Claim 2 (depends on 1) The process according to claim 1, wherein said stimulating additive is a nutrient. ### Claim 3 (depends on 1) The process according to claim 1, wherein said stimulating additive is selected from the group consisting of a nucleobase, a yeast extract and formic acid and/or a salt and/or ester thereof. ### Claim 4 (depends on 1) The process according to claim 1, wherein the starter culture is a mixed culture. ### Claim 5 (depends on 1) The process according to claim 1, wherein said microorganism is a fungus or a bacterium. ### Claim 6 (depends on 5) The process according to claim 5, wherein the bacterium is a lactic acid bacterium. ### Claim 7 (depends on 1) The process according to claim 1, wherein said process further comprises packing the starter culture composition. ### Claim 8 (depends on 1) The process according to claim 1, wherein an additive selected from the group consisting of an excipient, a carrier and a nutrient is added to said microorganism before freezing. ### Claim 9 A frozen starter culture composition comprising: a) frozen pellets or frozen granules comprising a microorganism and a cryoprotectant; and b) at least one frozen stimulating additive in the form of pellets or granules; wherein said at least one stimulating additive is selected from the group consisting of antioxidants, peptones, tryptones, vitamins, hydrolysates, amino acids, peptides, proteins, nucleic acids, nucleotides, nucleobases, yeast extracts, beef extract, organic acids, and growth factors; wherein no stimulating additive is present in the frozen pellets or granules comprising the microorganism and cryoprotectant, wherein the cryoprotectant is present in an amount effective for protection of the microorganism from damage during freezing and thawing and to increase the acidification rate of the starter culture composition, as compared to a composition without the cryoprotectant; and wherein the ratio of stimulating additive (wt.) to microorganism (wt.) in the starter culture composition is between 0.1 and 10; and wherein the at least one frozen stimulating additive increases the acidification rate of the starter culture composition as compared to a composition where a stimulating additive is frozen together with the microorganism and cryoprotectant. ### Claim 10 (depends on 9) A kit comprising at least a first packaging comprising the frozen pellets or frozen granules comprising a microorganism and a cryoprotectant according to claim 9 and a second packaging comprising the at least one frozen stimulating additive in the form of pellets or granules according to claim 9. ### Claim 11 (depends on 9) A process of producing a cultured food product, said process comprising: a. inoculating a precursor material with a frozen starter culture composition as defined in claim 9, b. culturing the precursor material with the frozen starter culture composition, and c. producing the cultured food product. ### Claim 12 (depends on 11) The process according to claim 11, wherein said cultured food product is selected from the group consisting of a dairy food product, a dairy food-derived product, a fermented milk product, a meat product, a vegetable product, a beverage product, a wine product, a bakery product, and a fruit juice product. ### Claim 13 (depends on 11) The process according to claim 11, wherein said precursor material is a dairy substrate. ### Claim 14 (depends on 13) The process according to claim 13, wherein said dairy substrate is milk.