Salt formulations for the fermentation of marine microorganisms

Abstract

Growth medium are disclosed for use in fermenting a marine microorganism. The medium comprise Potassium, Calcium, Strontium, Borate and Fluoride at specific concentrations. Alternatively, the growth medium comprises cobalt at specified concentrations or comprises vitamin B12 at specified concentrations. Methods of producing certain desired compound by fermentation of a marine microorganism are also disclosed.

1 .- 29 . (canceled) 
     
     
         30 . A method of producing Salinosporamide A, comprising the steps of
 (i) preparing a microbial growth medium;   (ii) inoculating said microbial growth medium with a strain of microorganism capable of producing Salinosporamide A;   (iii) adding a resin to the microbial growth medium before 96 hours after inoculation of the media,   
       wherein the growth medium comprises CaCO 3 , Fe 2 (SO 4 ) 3 , and KBr, and wherein the final concentration of the resin in the growth medium is 20 g/L, 30 g/L, or 40 g/L. 
     
     
         31 . The method of  claim 30 , wherein the resin is added before 24 hours, before 48 hours, or before 72 hours after inoculation of the microbial growth medium. 
     
     
         32 . (canceled) 
     
     
         33 . The method of  claim 30 , wherein the microorganism is of the species  Salinispora.    
     
     
         34 . The method of  claim 30 , wherein the microorganism is  Salinispora tropica.    
     
     
         35 . (canceled) 
     
     
         36 . The method of  claim 30 , wherein the resin is an adsorption resin. 
     
     
         37 . The method of  claim 30 , wherein the resin is a hydrophobic adsorption resin. 
     
     
         38 . The method of  claim 30 , wherein the resin includes at least one resin selected from the group consisting of Amberlite XAD2, Amberlite XAD4, Amberlite XAD7, Amberlite XAD7HP, Amberlite XAD16, Amberlite XAD761, Amberlite XAD 761, Amberlite XAD1180, Diaion HP2MG, Diaion HP20, Diaion HP21, Sepabeads SP825, Sepabeads SP850, Sepabeads SP70, Sepabeads SP700, and Sepabeads SP207. 
     
     
         39 . The method of  claim 30 , wherein the resin includes at least one resin selected from the group consisting of Amberlite XAD2, Amberlite XAD4, Amberlite XAD7, Amberlite XAD7HP and Amberlite XAD16. 
     
     
         40 .- 48 . (canceled) 
     
     
         49 . A method of producing a compound of Formula A; 
       
         
           
           
               
               
           
         
       
       wherein R 1  is selected from the group consisting of hydrogen and hydroxide, R2 is selected from the group consisting of 2-chloroethyl, 2-bromoethyl, ethyl, methyl, and propyl, and R3 is selected from the group consisting of methyl and ethyl, comprising the steps of growing a culture of  S. tropica  in the presence of XAD-7 resin. 
     
     
         50 . The method of  claim 49 , wherein the compound of Formula A has the following structure: 
       
         
           
           
               
               
           
         
       
     
     
         51 . The method of  claim 49 , wherein the compound of Formula A has the following structure: 
       
         
           
           
               
               
           
         
       
       wherein R 2  is selected from the group consisting of 2-chloroethyl and methyl. 
     
     
         52 .- 59 . (canceled) 
     
     
         60 . The method of  claim 30 , wherein the final concentration of the resin in the growth medium is 30 g/L. 
     
     
         61 . The method of  claim 30 , wherein the microbial growth medium comprises CaCO 3  at a concentration of between 2 mg/L and 50 g/L, Fe 2 (SO 4 ) 3  at a concentration of between 0.5 mg/L and 10 g/L, and KBr at a concentration of between 0.5 mg/L and 10 g/L. 
     
     
         62 . The method of  claim 30 , wherein the microbial growth medium further comprises a nitrogen source and a carbon source. 
     
     
         63 . The method of  claim 49 , wherein the culture of  S. tropica  is grown in a microbial growth medium. 
     
     
         64 . The method of  claim 63 , wherein the microbial growth medium comprises CaCO 3 , Fe 2 (SO 4 ) 3 , and KBr. 
     
     
         65 . The method of  claim 64 , wherein the microbial growth medium comprises CaCO 3  at a concentration of between 2 mg/L and 50 g/L, Fe 2 (SO 4 ) 3  at a concentration of between 0.5 mg/L and 10 g/L, and KBr at a concentration of between 0.5 mg/L and 10 g/L. 
     
     
         66 . The method of  claim 64 , wherein the microbial growth medium further comprises a nitrogen source and a carbon source. 
     
     
         67 . The method of  claim 49 , wherein the final concentration of the resin in the growth medium is 20 g/L, 30 g/L, or 40 g/L. 
     
     
         68 . A salt formulation for use in fermenting a marine microorganism, the formulation comprising:
 KCl at a mass percent between 50% and 70%;   CaCl 2  at a mass percent between 30% and 45%;   SrCl 2  at a mass percent between 1% and 3%;   H 3 BO 3  at a mass percent between 1% and 3%; and   NaF at a mass percent between 0.1% and 0.3%.