Neurotransmitters and Methods of Making the Same

Abstract

In an aspect, the disclosure provides methods for making neurotransmitters in a host organism. The neurotransmitters can be cannabinoids and derivatives of cannabinoids. The host cells can be microalgae, fungi or other host cells. In a related aspect, the disclosure provides host cells engineered to have biochemical pathways for making neurotransmitters such as cannabinoids.

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         21 . A method, comprising the steps of: providing a Chlorella microalgae host cell wherein the Chlorella microalgae host cell comprises a first nucleic acid encoding a hexanoyl-CoA synthase from  Cannabis sativa,  a second nucleic acid encoding a 3,5,7-trioxododecanoyl-CoA synthase from  Cannabis sativa , a third nucleic acid encoding an olivetolic acid cyclase from  Cannabis sativa , and a fourth nucleic acid encoding a geranyl-diphosphate:olivetolate geranyltransferase from  Cannabis sativa , wherein the first nucleic acid is operably linked to a control region, wherein the second nucleic acid is operably linked to a control region, wherein the third nucleic acid is operably linked to a control region, and wherein the fourth nucleic acid is operably linked to a control region, introducing the microalgae cell into a media wherein the media comprises a carbon source, culturing the microalgae cell in the media whereby a cannabinoid is made. 
     
     
         22 . The method of  claim 1 , wherein a carboxylic acid is made by the microalgae from the carbon source. 
     
     
         23 . The method of  claim 1 , wherein the carbon source is a carboxylic acid. 
     
     
         24 . The method of  claim 1 , wherein the carbon source is a hexanoic acid, and the cannabinoid is a cannabigerolic acid. 
     
     
         25 . The method of  claim 1 , wherein the carbon source is a butyric acid, and the cannabinoid is a cannabigerovarinic acid. 
     
     
         26 . The method of  claim 1 , wherein the microalgae further comprises a fifth nucleic acid encoding a cannabichromenic acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         27 . The method of  claim 6 , wherein the carbon source is a hexanoic acid, and the cannabinoid is a cannabigerolic acid and a cannabichromenic acid. 
     
     
         28 . The method of  claim 6 , wherein the carbon source is a butyric acid, and the cannabinoid is a cannabigerovarinic acid and a cannabichromevarinic acid. 
     
     
         29 . The method of  claim 1 , wherein the microalgae further comprises a fifth nucleic acid encoding a cannabidiolic-acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         30 . The method of  claim 1 , wherein the carbon source is a hexanoic acid, and the cannabinoid is a cannabigerolic acid and a cannabidiolic acid, and wherein the microalgae further comprises a fifth nucleic acid encoding a cannabidiolic-acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         31 . The method of  claim 1 , wherein the carbon source is a butyric acid, and the cannabinoid is a cannabigerovarinic acid and a cannabidivarinic acid, and wherein the microalgae further comprises a fifth nucleic acid encoding a cannabidiolic-acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         32 . The method of  claim 1 , wherein the microalgae further comprises a fifth nucleic acid encoding a Δ1-tetrahydrocannabinolic acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         33 . The method of  claim 1 , wherein the carbon source is a hexanoic acid, and the cannabinoid is a cannabigerolic acid and a tetrahydrocannabinolic acid, and wherein the microalgae further comprises a fifth nucleic acid encoding a Δ1-tetrahydrocannabinolic acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         34 . The method of  claim 1 , wherein the carbon source is a butyric acid, and the cannabinoid is a cannabigerovarinic acid and a tetrahydrocannabivarinic acid, and wherein the microalgae further comprises a fifth nucleic acid encoding a Δ1-tetrahydrocannabinolic acid synthase from  Cannabis sativa , and wherein the fifth nucleic acid is operably linked to a control region. 
     
     
         35 . The method of  claim 6 , wherein the microalgae further comprises a sixth nucleic acid encoding a cannabidiolic-acid synthase from  Cannabis sativa , and wherein the sixth nucleic acid is operably linked to a control region. 
     
     
         36 . The method of  claim 1 , wherein the carbon source is a hexanoic acid, and the cannabinoid is a cannabigerolic acid, a cannabichrominic acid and a cannabidiolic acid, and wherein the microalgae further comprises a sixth nucleic acid encoding a cannabidiolic-acid synthase from  Cannabis sativa , and wherein the sixth nucleic acid is operably linked to a control region. 
     
     
         37 . The method of  claim 1 , wherein the carbon source is a butyric acid, and the cannabinoid is a cannabigerovarinic acid, a cannabichromevarinic acid and a cannabidivarinic acid, and wherein the microalgae further comprises a sixth nucleic acid encoding a cannabidiolic-acid synthase from  Cannabis sativa , and wherein the sixth nucleic acid is operably linked to a control region. 
     
     
         38 . The method of  claim 15 , wherein the microalgae further comprises a seventh nucleic acid encoding a Δ1-tetrahydrocannabinolic acid synthase from  Cannabis sativa , and wherein the seventh nucleic acid is operably linked to a control region. 
     
     
         39 . The method of  claim 1 , wherein the carbon source is a hexanoic acid, and the cannabinoid is a cannabigerolic acid, a cannabichromenic acid, a cannabidiolic acid, and a tetrahydrocannabinolic acid, and wherein the microalgae further comprises a seventh nucleic acid encoding a Δ1-tetrahydrocannabinolic acid synthase from  Cannabis sativa , and wherein the seventh nucleic acid is operably linked to a control region. 
     
     
         40 . The method of  claim 1 , wherein the carbon source is a butyric acid, and the cannabinoid is a cannabigerovarinic acid, a cannabichromevarinic acid, a cannabidivarinic acid, and a tetrahydrocannabivarinic acid, and wherein the microalgae further comprises a seventh nucleic acid encoding a Δ1-tetrahydrocannabinolic acid synthase from  Cannabis sativa , and wherein the seventh nucleic acid is operably linked to a control region.