Chemical engineering processes and apparatus for the synthesis of compounds

Abstract

The present invention provides methods for producing cannabinoids and cannabinoid analogs as well as a system for producing these compounds. The inventive method is directed to contacting a compound according to Formula I or Formula II with a cannabinoid synthase. Also described is a system for producing cannabinoids and cannabinoid analogs by contacting a THCA synthase with a cannabinoid precursor and modifying at least one property of the reaction mixture to influence the quantity formed of a first cannabinoid relative to the quantity formed of a second cannabinoid.

1 . A system for producing cannabinoid products, comprising:
 a fermentor holding a cell culture medium comprising genetically engineered yeast or bacterial cells that are (a) producing a cannabinoid synthase selected from the group consisting of tetrahydrocannabinolic acid (THCA) synthase and cannabidiolic acid (CBDA) synthase and (b) secreting the cannabinoid synthase into the culture medium; and   a bioreactor containing a reaction mixture comprising the cannabinoid synthase and 3-(3,7-dimethylocta-2,6-diene-1-yl)-2,4-dihydroxy-6-propyl-1-benzoic acid (CBGVA) and configured to form one or more cannabinoid products.   
     
     
         2 . The system of  claim 1 , further comprising a control mechanism configured to control one or more properties of the reaction mixture so as to modify the amount of the one or more cannabinoid products, wherein the one or more properties of the reaction mixture are selected from the group consisting of ionic strength, pH, temperature, and pressure. 
     
     
         3 . The system of  claim 2 , wherein the property of the reaction mixture is pH. 
     
     
         4 . The system of  claim 3 , wherein the pH is from 4 to less than 8. 
     
     
         5 . The system of  claim 4 , wherein the pH is between 4.0 and 6.0. 
     
     
         6 . The system of  claim 2 , wherein the control mechanism comprises a processing circuit and a display to control the one or more properties of the reaction mixture. 
     
     
         7 . The system of  claim 1 , further comprising a control mechanism configured to regulate a property of the culture medium containing the cannabinoid synthase, wherein the property of the culture medium containing the cannabinoid synthase is one or more of oxygen level of the culture medium, rate of agitation of the culture medium, pH of the culture medium, or flow rate of the culture medium into the bioreactor. 
     
     
         8 . The system of  claim 1 , further comprising a filter configured to separate the cells from the culture medium containing the cannabinoid synthase produced by the cells. 
     
     
         9 . The system of  claim 1 , wherein the cannabinoid synthase is conjugated to a solid support in the bioreactor. 
     
     
         10 . The system of  claim 1 , wherein the bacterial cells are  Escherichia coli  cells. 
     
     
         11 . The system of  claim 1 , wherein the reaction mixture comprises a solvent. 
     
     
         12 . The system of  claim 11 , wherein the solvent is one or more of dimethyl sulfoxide (DMSO), dimethyl formamide (DMF), iso-propyl alcohol, and cyclodextrin. 
     
     
         13 . The system of  claim 12 , wherein the concentration of the solvent in the reaction mixture is between 5% and 30% (w/v). 
     
     
         14 . The system of  claim 1 , wherein the cells are genetically modified to produce and secrete THCA synthase. 
     
     
         15 . The system of  claim 1 , wherein the cells are genetically modified to produce and secrete CBDA synthase. 
     
     
         16 . A method for producing a tetrahydrocannabinol, cannabichromene, or both tetrahydrocannabinol and cannabichromene, or their analogs comprising the steps of:
 (a) selecting a compound according to Formula V:   
       
         
           
           
               
               
           
         
         (b) reacting the Formula V compound with THCA synthase in the presence of a solvent; 
         (c) modifying at least one property of a reaction mixture comprising the Formula V compound and THCA synthase to obtain a tetrahydrocannabinol, a cannabichromene, or both tetrahydrocannabinol and cannabichromene, or their analogs as products; 
         wherein 
         R is selected from —OH, halogen, —SH, or a —NR a R b  group; 
         R 1  and R 2  are each independently selected from the group consisting of —H, —C(O)R a , —OR a , an optionally substituted C 1 -C 10  linear or branched alkylene, an optionally substituted C 2 -C 10  linear or branched alkenylene, an optionally substituted C 2 -C 10  linear or branched alkynylene, an optionally substituted C 3 -C 10  aryl, an optionally substituted C 3 -C 10  cycloalkyl, (C 3 -C 10 )aryl-(C 1 -C 10 )alkylene, (C 3 -C 10 )aryl-(C 2 -C 10 )alkenylene, and (C 3 -C 10 )aryl-(C 1 -C 10 )alkynylene, or 
         R 1  and R 2  together with the carbon atoms to which they are bonded form a C 5 -C 10  cyclic ring; 
         R 3  is selected from the group consisting of H, —C(O)R a , and C 1 -C 10  linear or branched alkyl; and 
         R a  and R b  are each independently selected from the group consisting of —H. —OH, —SH, —NH 2 , (C 1 -C 10 ) linear or branched alkyl, and a C 3 -C 10  cycloalkyl. 
       
     
     
         17 . The method of  claim 16 , wherein step (c) comprises changing the pH of the reaction mixture. 
     
     
         18 . The method of  claim 17 , wherein the pH of the reaction mixture is from about 4.0 to about 8.0. 
     
     
         19 . A method for optimizing production of one or more cannabinoids comprising:
 (a) cloning cannabinoid synthases from multiple sources into host cells, expressing the cannabinoid synthases in the host cells, and isolating the cannabinoid synthases from the host cells;   (b) determining the catalytic activity of each of the isolated cannabinoid synthases and comparing the catalytic activities to identify an optimal cannabinoid synthase having a high catalytic activity;   (c) inserting multiple copies of the optimal cannabinoid synthase into an expression vector;   (d) transforming a host cell with the expression vector obtained in step (c), expressing the optimal cannabinoid synthase in the transformed host cell, and isolating the optimal cannabinoid synthase from the transformed host cell; and   (e) reacting cannabigerolic acid (CBGA) or CBGVA with the optimal cannabinoid synthase isolated in step (d) in a reaction mixture to produce one or more cannabinoids;   wherein the optimal cannabinoid synthase is a CBDA synthase or a THCA synthase, and wherein the reaction mixture comprises one or more solvents of DMSO, DMF, iso-propyl alcohol, and cyclodextrin.   
     
     
         20 . The method of  claim 19 , wherein the amount of the solvent in the reaction mixture is between 5% and 30% (w/v), and wherein the pH of the reaction mixture is between 4.0 and 8.0.