Compositions and methods for the production of compounds
AssigneeGinkgo Bioworks, Inc.
InventorsDaniel C. GRAY, Enhu LI, Brian R. BOWMAN, Gregory L. VERDINE, Keith ROBISON, Marc CHEVRETTE, Dan UDWARY, Pam Shouping WANG, Anna LI, Jay P. MORGENSTERN
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Abstract
The present disclosure provides proteins, nucleic acids, vectors, and host molecules useful for the production of compounds of interest, and methods for their use.
Claims51
An engineered polyketide synthase comprising one or more heterologous modules with altered enzymatic activity relative to a reference polyketide, wherein the engineered polyketide synthase is capable of producing a polyketide when expressed under conditions suitable to allow expression of a compound by the engineered polyketide synthase and wherein the one or more heterologous modules do not substantially inhibit polyketide translocation during polyketide biosynthesis.
An engineered polyketide synthase comprising one or more heterologous modules with altered enzymatic activity relative to a reference polyketide, wherein the engineered polyketide synthase is capable of producing a polyketide when expressed under conditions suitable to allow expression of a compound by the engineered polyketide synthase and wherein the one or more heterologous modules comprise linking sequences which are compatible to the linking sequences of the modules adjacent thereto.
An engineered polyketide synthase comprising one or more heterologous modules with altered enzymatic activity relative to a reference polyketide, wherein the engineered polyketide synthase is capable of producing a polyketide when expressed under conditions suitable to allow expression of a compound by the engineered polyketide synthase and wherein the polyketide expression level of the engineered polyketide synthase is at least 1% of the polyketide expression level of the reference polyketide synthase.
A chimeric polyketide synthase, wherein at least one module of the chimeric polyketide synthase has been modified as compared to a polyketide synthase having the sequence of SEQ ID NO:
- 175-176.
The nucleic acid of claim 15, wherein the nucleic acid further encodes an LAL, wherein the sequence encoding the LAL is operatively linked to the sequence encoding the polyketide synthase.
A method of producing a compound, the method comprising:
- a) providing a parent polyketide synthase sequence capable of producing a compound;
- (b) determining the compatibility of at least one module of a second polyketide synthase with at least two modules of the parent polyketide synthase;
- (c) producing a nucleic acid encoding a modified polyketide synthase, wherein the modified polyketide synthase comprises at least one module of a second polyketide synthase which has been determined to be compatible with the at least two modules of the parent polyketide synthase.
A method of producing a compound, the method comprising:
- (a) providing a parent nucleic acid encoding a parent polyketide synthase;
- (b) modifying the parent nucleic acid to create a modified nucleic acid encoding a modified polyketide synthase capable of producing a compound, wherein the modification produces a modified polyketide synthase comprising at least one heterologous module.
A method of producing a compound, the method comprising:
- (a) providing a parent polynucleotide sequence capable of producing a compound;
- (b) identifying one or more heterologous modules suitable for replacement of one or more modules in the parent polynucleotide sequence;
- (c) producing a nucleic acid encoding a modified polyketide synthase, wherein the modified polyketide synthase comprises at least one heterologous module identified in step (b).
A method of producing a plurality of polynucleotides, wherein each of the plurality of polynucleotides corresponds to an engineered polyketide synthase, and wherein each of the plurality of polynucleotides comprises one or more heterologous modules with altered enzymatic activity relative to a reference polyketide, wherein the method comprises:
- (a) providing a parent polynucleotide sequence encoding a polyketide synthase;
- (b) identifying one or more modules for replacement in the parent polynucleotide sequence;
- (c) identifying two or more heterologous modules suitable for replacement for each of the modules identified in step (b);
- (d) generating a plurality of polynucleotides, wherein each of the plurality of polynucleotides corresponds to an engineered polyketide synthase, and wherein each of the plurality of polynucleotides comprises a heterologous module selected from the two or more heterologous modules identified in step (c) in replacement of each of the one or more modules to be replaced identified in step (b).
Dependencies are inferred from the claim text.