System and Method for Liquid Culture of Anaerobic or Microaerophilic Microorganisms

Abstract

A culture system for culturing a microaerophilic or an anaerobic microorganism is provided. The culture system can include effective amounts of i) an enzyme of an oxidoreductase family and ii) a substrate for said enzyme, a container, and a predetermined volume of aqueous medium that supports growth of said anaerobic or microaerophilic microorganism. The enzyme can be selected from a group consisting of ascorbic acid oxidase and laccase. The effective amounts are effective to deplete dissolved oxygen in the predetermined volume to a concentration that facilitates growth of a microaerophilic microorganism or an obligately-anaerobic microorganism. A method of using the system is also provided.

1 - 13 . (canceled) 
     
     
         14 . A culture system for culturing a microaerophilic or an anaerobic microorganism, said culture system comprising:
 effective amounts of i) an enzyme of an oxidoreductase family and ii) a substrate for said enzyme, wherein the enzyme is selected from a group consisting of ascorbic acid oxidase and laccase;   a container; and   a predetermined volume of aqueous medium that supports growth of said anaerobic or microaerophilic microorganism;   wherein the effective amounts are effective to deplete dissolved oxygen in the predetermined volume to a concentration that facilitates growth of a microaerophilic microorganism or an obligately-anaerobic microorganism.   
     
     
         15 - 16 . (canceled) 
     
     
         17 . A kit, comprising:
 a sealable container; and   an oxygen scavenging system comprising i) an enzyme of an oxidoreductase family and ii) a substrate for said enzyme, wherein the enzyme is selected from a group consisting of ascorbic acid oxidase and laccase.   
     
     
         18 . The kit of  claim 17 , wherein the enzyme is allocated in one or more first vessel, each first vessel having an amount of enzyme effective react with the enzyme substrate to deplete dissolved oxygen in a predetermined volume of aqueous medium to a dissolved oxygen concentration that facilitates the growth of an anaerobic microorganism or a microaerophilic microorganism. 
     
     
         19 . The kit of  claim 17 , wherein the substrate is allocated in the one or more first vessel or one or more second vessel, each first vessel or second vessel having an amount of substrate effective to react with the enzyme to deplete dissolved oxygen in a predetermined volume of aqueous medium to a dissolved oxygen concentration that facilitates the growth of an anaerobic microorganism or a microaerophilic microorganism. 
     
     
         20 . (canceled) 
     
     
         21 . The kit of  claim 19 , wherein the first vessel and/or the second vessel is the container. 
     
     
         22 . The kit of  claim 17 , wherein the container is made of:
 glass; or   a polymeric material selected from a group consisting of polyvinyl chloride, polyethylene, polycarbonate, polystyrene, polyvinyledene chloride, polyacrylonitriles, ethylene vinyl acetate copolymers, polyethylene vinyl acetate copolymer, polyvinylidene chloride copolymer, ethylene vinyl alcohol copolymer, polyamide, polymethacrylonitrile, cellulose acetate, cellulose acetate butyrate, cellulose diacetate, Neoprene®, Teflon®, polysiloxane or mixtures or laminates thereof; or   a polymeric film comprising an oriented polymeric film selected from a group consisting of oriented polypropylene, oriented nylon, or oriented polyethylene terephthalate; or   a polymeric film comprising a metalized film; or   a polymeric film comprising polyvinyledene chloride, polyvinyledene chloride copolymer or polyvinyl alcohol coated film selected from a group consisting of polyvinyl alcohol coated oriented polypropylene, polyvinyledene chloride coated oriented polyethylene terephthalate, polyvinyledene chloride copolymer coated oriented polyethylene terephthalate, polyvinyledene chloride coated oriented polypropylene, or polyvinyledene chloride copolymer oriented polypropylene.   
     
     
         23 . The kit of  claim 17 , wherein the container is resealable. 
     
     
         24 . The kit of  claim 17 , wherein the container is single-use or disposable. 
     
     
         25 . The kit of  claim 17 , wherein the container is flexible. 
     
     
         26 . The kit of  claim 17 , wherein the container is selected from a group consisting of a pouch, an envelope, a sealed wrapping, a bag, a can, a jar, a barrel, a box, a barrier bag, a zippered bag, a storage bag, a dense lab bag, a flask, a flask with stopper, a bottle, a bottle with stopper, a tube, or a multi-layer polymeric film laminate. 
     
     
         27 . The kit of  claim 17 , wherein the container is sealed using a seal selected from a group consisting of a stopper, a septum, a screwcap, or a heat seal. 
     
     
         28 . The kit of  claim 17 , wherein the container comprises a sealable port. 
     
     
         29 . The kit of  claim 17 , wherein the container has an oxygen permeability of less than about 100 nmol/(m×s×Gpa). 
     
     
         30 . The kit of  claim 17 , wherein the container has an oxygen permeability of from about 6 to about 8 nmol/(m×s×Gpa). 
     
     
         31 . The kit of  claim 17 , wherein the container has a thickness of about 0.025 mm. 
     
     
         32 . The kit of  claim 17 , wherein the container has a headspace volume of about 0 mm to about 50 mm. 
     
     
         33 . The kit of  claim 17 , wherein the kit further comprises a material that can be placed into the container to indicate the presence or absence of oxygen in the container.