Production of ent-kaurene from lignocellulosic hydrolysate in Rhodosporidium toruloides

Table 1 Overview of constructs used to engineer ent-kaurene production in R. toruloides

Plasmids	Genotypes/features	ABFPUB registry ID
Construct 1	P_ANT-GfKS-HYG^R	ABFPUB_1
Construct 2	P_GAPDH-GfKS-NAT^R	ABFPUB_3
Construct 3	P_TEF1-RtERG20(F91C)-KanMX^R	ABFPUB_6
Construct 4	P_TEF1-TcGGPPS(S239C,G29D)-KanMX^R	ABFPUB_7
Construct 5	P_TEF1-GgFPS(F112A)-KanMX^R	ABFPUB_8
Construct 6	P_TEF1-RtBTS1-KanMX^R	ABFPUB_9
Construct 7	P_ANT-GfKS-P_TEF1-RtERG20(F91C)-NAT^R	ABFPUB_10
Construct 8	P_ANT-GfKS-P_TEF1-TcGGPPS(S239C, G295D)-NAT^R	ABFPUB_11
Construct 9	P_ANT-GfKS-P_TEF1-GgFPS(F112A)-NAT^R	ABFPUB_12
Construct 10	P_ANT-GfKS-P_TEF1-RtBTS1-NAT^R	ABFPUB_13
Strains	IFO0880 (WT)	ABFPUB_14
	IFO0880/P_GAPDH-GfKS-NAT^R	ABFPUB_15–17
	IFO0880/P_ANT-GfKS-HYG^R	ABFPUB_18, 30, 34
	IFO0880/P_GAPDH-GfKS-NAT^R/P_TEF1-RtERG20(F91C)-KanMX^R	ABFPUB_47–49
	IFO0880/P_GAPDH-GfKS-NAT^R/P_TEF1-TcGGPPS(S239C,G29D)-KanMX^R	ABFPUB_19, 20, 31
	IFO0880/P_GAPDH-GfKS-NAT^R/P_TEF1-GgFPS(F112A)-KanMX^R	ABFPUB_32, 33, 35
	IFO0880/P_GAPDH-GfKS-NAT^R/P_TEF1-RtBTS1-KanMX^R	ABFPUB_38–40
	IFO0880/P_ANT-GfKS-P_TEF1-RtERG20(F91C)-NAT^R	ABFPUB_41–43
	IFO0880/P_ANT-GfKS-P_TEF1-TcGGPPS(S239C,G295D)-NAT^R	ABFPUB_21, 22, 29
	IFO0880/P_ANT-GfKS-P_TEF1-GgFPS(F112A)-NAT^R	ABFPUB_23, 26, 36
	IFO0880/P_ANT-GfKS-P_TEF1-RtBTS1-NAT^R	ABFPUB_44–46

Strains and plasmids used in this study are available upon request through the Agile BioFoundry Strain Registry (http://public-registry.agilebiofoundry.org// [37])
GAPDH glyceraldehyde 3-phosphate dehydrogenase, ANT, adenine nucleotide translocase, TEF1 translational elongation factor, GfKS, Gibberella fujikuroi kaurene synthase (NCBI Accession Number, Q9UVY5), RtERG20(F91C) mutant farnesyl pyrophosphate synthase from R. toruloides (PRQ75922.1), TcGGPPS(S239C,G295D) mutant geranylgeranyl pyrophosphate synthase from Taxus canadensis (AAD16018), GgFPS(F112A) mutant farnesyl pyrophosphate synthase from Gallus gallus (P08836.2) and RtBTS1 geranylgeranyl pyrophosphate synthase BTS1 from R. toruloides (PRQ72579)

ISSN: 1475-2859