Method for producing terpenes
AssigneeTeknologian tutkimuskeskus VTT Oy
InventorsKirsi Bromann, Mervi Toivari, Tiina Nakari-Setälä, Laura Ruohonen
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Abstract
The present invention concerns a method for producing terpenes in fungi comprising the steps of (a) providing a modified terpene biosynthetic gene cluster inside a host cell, wherein one or more of the naturally occurring genes or promoters of the cluster have been replaced, truncated or removed, (b) providing a transcription factor inside the host cell, the transcription factor activating the terpene biosynthetic gene cluster; (c) cultivating said host in conditions allowing the expression of the transcription factor activating the cluster; and optionally (d) recovering the thus produced terpene product.
Claims28
Method for producing terpenes in fungi comprising the steps of:
- providing a modified terpene biosynthetic gene cluster inside a host cell, the gene cluster having naturally occurring terpene biosynthetic genes and promoters, wherein one or more of these genes or promoters have been replaced, truncated or removed,
- providing a transcription factor inside the host cell so that the transcription factor is operably linked to one of the natural or modified promoters, the transcription factor activating the terpene biosynthetic gene cluster having terpene biosynthetic genes and regulatory regions operably linked to said genes;
- and
- cultivating said host in conditions allowing the expression of the transcription factor activating the cluster.
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A terpene biosynthetic gene cluster comprising the genes putatively encoding (a) Zn(II)2Cys6-type transcription factor (AN1599), monoterpene synthase operably linked to the regulatory regions of AN1594, HMG-CoA reductase (AN1593), GGPP-synthase (AN1592) and
- (b) optionally translation elongation factor 1-gamma (AN1595), cytochrome P450 (AN1598), short-chain dehydrogenase (AN1596), hypothetical protein with some similarity to methyltransferase (AN1597), the regulatory regions operably linked to said genes, and optionally an AAA family ATPase (AN1591) and
- (c) regulatory regions operably linked to the genes of item (a) and to the genes of item (b).
A terpene biosynthetic gene cluster comprising the genes putatively encoding
- (a) Zn(II)2Cys6-type transcription factor (AN1599), monoterpene synthase operably linked to the regulatory regions of AN1594, HMG-CoA reductase (AN1593), GPP-synthase operably linked to the regulatory regions of AN1592 and
- (b) optionally translation elongation factor 1-gamma (AN1595), cytochrome P450 (AN1598), short-chain dehydrogenase (AN1596), hypothetical protein with some similarity to methyltransferase (AN1597), the regulatory regions operably linked to said genes, and optionally an AAA family ATPase (AN1591) and
- (c) regulatory regions operably linked to the genes of item (a) and to the genes of item (b).
A terpene biosynthetic gene cluster comprising the genes putatively encoding
- (a) Zn(II)2Cys6-type transcription factor (AN1599), sesquiterpene synthase operably linked to the regulatory regions of AN1594, HMG-CoA reductase (AN1593), GGPP-synthase (AN1592) and
- (b) optionally translation elongation factor 1-gamma (AN1595), cytochrome P450 (AN1598), short-chain dehydrogenase (AN1596), hypothetical protein with some similarity to methyltransferase (AN1597), the regulatory regions operably linked to said genes, and optionally an AAA family ATPase (AN1591) and
- (c) regulatory regions operably linked to the genes of item (a) and to the genes of item (b).
A terpene biosynthetic gene cluster comprising
- (a) Zn(II)2Cys6-type transcription factor (AN1599), sesquiterpene synthase operably linked to the regulatory regions of AN1594, HMG-CoA reductase (AN1593), FPP-synthase operably linked to the regulatory regions of AN1592 and
- (b) optionally translation elongation factor 1-gamma (AN1595), cytochrome P450 (AN1598), short-chain dehydrogenase (AN1596), hypothetical protein with some similarity to methyltransferase (AN1597), the regulatory regions operably linked to said genes, and optionally an AAA family ATPase (AN1591) and
- (c) regulatory regions operably linked to the genes of item (a) and to the genes of item (b).
A terpene biosynthetic gene cluster comprising the genes putatively encoding
- (a) Zn(II)2Cys6-type transcription factor (AN1599), diterpene synthase operably linked to the regulatory regions of AN1594, HMG-CoA reductase (AN1593), GGPP-synthase (AN1592) and
- (b) optionally translation elongation factor 1-gamma (AN1595), cytochrome P450 (AN1598), short-chain dehydrogenase (AN1596), hypothetical protein with some similarity to methyltransferase (AN1597), the regulatory regions operably linked to said genes, and optionally an AAA family ATPase (AN1591) and
- (c) regulatory regions operably linked to the genes of item (a) and to the genes of item (b).
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A fungal production host, wherein a modified terpene biosynthetic gene cluster comprising the genes putatively encoding
- (a) Zn(II)2Cys6-type transcription factor (AN1599), monoterpene synthase operably linked to the regulatory regions of AN1594, HMG-CoA reductase (AN1593), GGPP-synthase (AN1592) and
- (b) optionally translation elongation factor 1-gamma (AN1595), cytochrome P450 (AN1598), short-chain dehydrogenase (AN1596), hypothetical protein with some similarity to methyltransferase (AN1597), the regulatory regions operably linked to said genes, and optionally an AAA family ATPase (AN1591) and
- (c) regulatory regions operably linked to the genes of item (a) and to the genes of item (b) includes a transcription factor having a sequence characterized by SEQ ID NO:
- 52 or a sequence showing at least 80% degree of identity to SEQ ID NO:
- 52, operably linked to a promoter.
Dependencies are inferred from the claim text.