# US 11,028,454 B2 · Incorporation of thermo-resistant and/or pressure-resistant organisms in materials > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,028,454 B2 - Document ID: US11028454B2 - Status: Granted - Kind: B2 - Application number: 13/120,908 - Filed: 2009-09-24 - Published: 2011-09-08 - Granted: 2021-06-08 - Earliest claimed: 2008-09-24 - Assignees: Resilux - Applicants: Johan Mertens; Lynda Beladjal; Frank Devlieghere; Sam Verbrugghe; Benedikt Sas; Filip Du Prez; Tom Anthierens; Assia Ouchchen - Inventors: Johan Mertens; Lynda Beladjal; Frank Devlieghere; Sam Verbrugghe; Benedikt Sas; Filip Du Prez; Tom Anthierens; Assia Ouchchen - Web page: https://patentgrep.com/patent/US11028454B2 ## CPC - C12N 1/205: Bacterial isolates - C12N 1/20: Bacteria; Culture media therefor - C12N 11/093: Polyurethanes - C12N 11/096: Polyesters; Polyamides - C12R 2001/07: Bacillus - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS ## Abstract The present application relates to methods for producing materials having viable micro-organisms incorporated therein. More particularly, the materials are produced under high temperature and/or high pressure conditions, and the micro-organisms are incorporated in the material before or during these conditions. Also provided are micro-organisms that remain viable under these conditions and have different applications, depending on the nature of the material. ## Claims ### Claim 1 An apparatus comprising a beverage bottle or food container made of a material comprising a polymer and a micro-organism powder comprising living or viable spores of a micro-organism incorporated into said polymer and prepared by a process comprising mixing a polymer matrix comprising said polymer and said micro-organism powder at a pressure ranging from 1.5 to 50 bar or at temperatures ranging from 100° C. to 220° C. at a pressure ranging from 1.5 to 50 bar, wherein said micro-organism is a Bacillus amyloliquefaciens strain deposited under No. 1D9698 at the Belgian Co-ordinated Collections of Micro-organisms (BCCM), or a Bacillus strain with a gyrA sequence that is at least 95% identical to SEQ ID NO:1, and which is capable of withstanding a pressure ranging from 1.5 to 50 bar or at temperatures ranging from 100° C. to 220° C. at a pressure ranging from 1.5 to 50 bar, and wherein said polymer comprises at least one monomer selected from the group consisting of acrylates, methacrylates, acrylamides, methacrylamides, alcohols, amines, anhydrides, epoxides, styrenes, functionalized vinyls, functionalized allyls, propenes, butadienes, ethylenes, isocyanates, lactams, lactones, saccharides, glucose and esters. ### Claim 2 (depends on 1) The apparatus according to claim 1 acting as an oxygen barrier. ### Claim 3 (depends on 1) The apparatus according to claim 1, wherein said gyrA sequence is at least 98% identical to SEQ ID NO:1. ### Claim 4 (depends on 1) The apparatus according to claim 1, wherein said gyrA sequence is identical to SEQ ID NO:1. ### Claim 5 (depends on 1) The apparatus according to claim 1, wherein said micro-organism is the Bacillus amyloliquefaciens strain deposited under No. ID9698 at the Belgian Co-ordinated Collections of Micro-organisms (BCCM). ### Claim 6 (depends on 3) The apparatus of claim 3 acting as an oxygen barrier. ### Claim 7 (depends on 5) The apparatus of claim 5 acting as an oxygen barrier. ### Claim 8 An apparatus comprising a beverage bottle or a food container made of a polymer and a micro-organism powder comprising living or viable spores of a micro-organism incorporated into said polymer and prepared by a process comprising mixing a polymer matrix comprising said polymer and said micro-organism powder at a pressure ranging from 1.5 to 50 bar or at temperatures ranging from 100° C. to 220° C. at a pressure ranging from 1.5 to 50 bar, wherein said micro-organism is a Bacillus amyloliquefaciens strain deposited under No. ID9698 at the Belgian Co-ordinated Collections of Micro-organisms (BCCM), or a Bacillus strain with a gyrA sequence that is at least 95% identical to SEQ ID NO:1, and which is capable of withstanding a pressure ranging from 1.5 to 50 bar or at temperatures ranging from 100° C. to 220° C. at a pressure ranging from 1.5 to 50 bar. ### Claim 9 (depends on 8) The apparatus of claim 8 acting as an oxygen barrier. ### Claim 10 (depends on 8) The apparatus according to claim 8, wherein said gyrA sequence is at least 98% identical to SEQ ID NO:1. ### Claim 11 (depends on 8) The apparatus according to claim 8, wherein said gyrA sequence is identical to SEQ ID NO:1. ### Claim 12 (depends on 8) The apparatus according to claim 8, wherein said micro-organism is the Bacillus amyloliquefaciens strain deposited under No. ID9698 at the Belgian Co-ordinated Collections of Micro-organisms (BCCM). ### Claim 13 (depends on 1) The apparatus according to claim 1, wherein the polymer has a polarity ranging from 17-21 (MPa)1/2. ### Claim 14 (depends on 1) The apparatus according to claim 1, wherein the polymer has a minimal water absorption value selected from values of 0.01 to 0.1. ### Claim 15 (depends on 8) The apparatus according to claim 8, wherein the polymer has a polarity ranging from 17-21 (MPa)1/2. ### Claim 16 (depends on 8) The apparatus according to claim 8, wherein the polymer has a minimal water absorption value selected from values of at least 0.01, 0.02, 0.1, or 0.2. ### Claim 17 An apparatus comprising a food and beverage bottle or container made of material comprising a polymer and a micro-organism powder comprising living or viable spores of a micro-organism incorporated into said polymer in a polymer matrix having a polarity ranging from 17-21 (MPa)1/2 and having minimal water absorption value ranging from of at least 0.01 to 0.2, wherein said apparatus is prepared by a process comprising mixing said polymer matrix and said micro-organism at pressure ranging from 1.5 to 50 bar or at temperatures ranging from 100° C. to 220° C. at a pressure ranging from 1.5 to 50 bar, wherein said micro-organism is a Bacillus strain with a gyrA sequence that is at least 95% identical to SEQ ID NO:1, and wherein said polymer comprises at least one monomer selected from the group consisting of acrylates, methacrylates, acrylamides, methacrylamides, alcohols, amines, anhydrides, epoxides, styrenes, functionalized vinyls, functionalized allyls, propenes, butadienes, ethylenes, isocyanates, lactams, lactones, saccharides, glucose and esters. ### Claim 18 (depends on 17) The apparatus according to claim 17, wherein the bottles and containers have a carbon dioxide barrier, wherein said micro-organisms consume of the bottles and containers carbon dioxide, or alternatively produce carbon dioxide that desorbs in content or acts as an osmotic barrier, wherein said micro-organisms are rehydrated, by using said polymer which is able to become water-saturated, to become sufficiently hydrated so that the living and viable spores therein are enabled to increase their water content and revert to metabolically active micro-organisms; wherein a polymer having a sufficiently high water absorption value is used; wherein the H2O absorption value of the polymer is at least 0.01; at least 0.02; at least 0.1 or at least 0.2. ### Claim 19 (depends on 17) The apparatus of claim 17 comprising a bottle made of polyethylene terephthalate (PET), wherein said micro-organisms are incorporated into the bottle an oxygen barrier. ### Claim 20 (depends on 17) The apparatus according to claim 17, wherein said gyrA sequence is at least 98% identical to SEQ ID NO:1. ### Claim 21 (depends on 17) The apparatus according to claim 17, wherein said gyrA sequence is identical to SEQ ID NO:1.