Compositions and methods for production of myrcene
AssigneeAmyris, Inc.
InventorsAndrew Main, Grzegorz Wojciechowski, Yue Yang, Lishan Zhao
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Abstract
Provided herein are compositions and methods for producing myrcene by culturing genetically modified microbial host cells that express a myrcene synthase and optionally a geranyl pyrophosphate synthase. Also provided herein are isolated nucleic acid molecules that encode myrcene synthase variants derived from the Ocimum species myrcene synthase, which comprise one or more amino acid substitutions that improve in vivo performance of myrcene synthase in genetically modified microbial host cells. Also provided herein are isolated myrcene synthase variants that exhibit an improved activity for converting geranyl diphosphate into myrcene.
Claims24
A fermentation composition comprising:
- (a) a genetically modified microbial host cell cultured in a culture medium, wherein the microbial host cell comprises a heterologous nucleic acid molecule encoding a myrcene synthase, wherein the myrcene synthase comprises a sequence that has at least about 70% sequence identity to SEQ ID NO:
- 2, and at least one variant amino acid residue at position 27, 28, 207, 213, 222, 342, 347, 381, 382, 389, 390, 401, 404, 428, 439, 466, 482, 484, 505, 514, 517, 524, 527, 528, 543, 544, or 552, wherein the positions are numbered with reference to SEQ ID NO:
- 2;
- and
- (b) monoterpenes produced from the microbial host cell, wherein the monoterpenes comprise myrcene as a major component and one or more co-products as minor components, wherein the one or more co-products comprise α-terpinene and γ-terpinene.
A fermentation composition comprising:
- (a) a genetically modified microbial host cell cultured in a culture medium, wherein the microbial host cell comprises a heterologous nucleic acid molecule encoding a myrcene synthase, wherein the myrcene synthase comprises a sequence that has at least about 70% sequence identity to SEQ ID NO:
- 2, and wherein the myrcene synthase comprises variant amino acid residues F381L, I404V, E528D, and A544S compared to SEQ ID NO:
- 2, wherein the positions are numbered with reference to SEQ ID NO:
- 2;
- (b) monoterpenes produced from the microbial host cell, wherein the monoterpenes comprise myrcene as a major component and one or more co-products as minor components, wherein the one or more co-products comprise α-terpinene and γ-terpinene.
A fermentation composition comprising:
- (a) a genetically modified microbial host cell cultured in a culture medium, wherein the microbial host cell comprises a heterologous nucleic acid molecule encoding a myrcene synthase, wherein the myrcene synthase comprises a sequence that has at least about 70% sequence identity to SEQ ID NO:
- 2, and wherein the myrcene synthase comprises variant amino acid residues F381L, I404V, E528D, and Q552R compared to SEQ ID NO:
- 2, wherein the positions are numbered with reference to SEQ ID NO:
- 2;
- (b) monoterpenes produced from the microbial host cell, wherein the monoterpenes comprise myrcene as a major component and one or more co-products as minor components, wherein the one or more co-products comprise α-terpinene and γ-terpinene.
Dependencies are inferred from the claim text.