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Toward production of jet fuel functionality in oilseeds: identification of FatB acyl-acyl carrier protein thioesterases and evaluation of combinatorial expression strategies in Camelina seeds.

Kim HJ, Silva JE, Vu HS, Mockaitis K, Nam JW, Cahoon EB - J. Exp. Bot. (2015)

Bottom Line: Expression of CpuFatB3 and CvFatB1 resulted in Camelina oil with capric acid (10:0), and CpuFatB4 expression conferred myristic acid (14:0) production and increased 16:0.Increases in lauric acid (12:0) and 14:0, but not 10:0, in Camelina oil and at the sn-2 position of triacylglycerols resulted from inclusion of a coconut lysophosphatidic acid acyltransferase specialized for MCFAs.Camelina lines presented here provide platforms for additional metabolic engineering targeting fatty acid synthase and specialized acyltransferases for achieving oils with high levels of jet fuel-type fatty acids.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.

No MeSH data available.


Related in: MedlinePlus

Fatty acid composition of the sn-2 position of seed oil TAG. Fatty acid composition of the sn-2 position of seed oil TAG in plants expressing FatB genes alone or in combination with CnLPAT as determined by lipase digestion-based analyses. The data represent averages of four biological replicates ±SD. (A) CpFatB2 and CpFatB2 with CnLPAT. (B) UcFatB1 and UcFatB1 with CnLPAT. (C) ChFatB2 and ChFatB2 with CnLPAT.
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Figure 5: Fatty acid composition of the sn-2 position of seed oil TAG. Fatty acid composition of the sn-2 position of seed oil TAG in plants expressing FatB genes alone or in combination with CnLPAT as determined by lipase digestion-based analyses. The data represent averages of four biological replicates ±SD. (A) CpFatB2 and CpFatB2 with CnLPAT. (B) UcFatB1 and UcFatB1 with CnLPAT. (C) ChFatB2 and ChFatB2 with CnLPAT.

Mentions: To increase MCFA levels further in Camelina seeds, FatB polypeptides were co-expressed with a coconut lysophosphatidic acid acyltransferase (CnLPAT) that has been shown to insert 12:0 at the sn-2 position of the TAG glycerol backbone (Table 1), leading to 12:0 accumulation in transgenic B. napus seeds (Knutzon et al., 1999). Co-expression of either CpFatB2 or UcFatB1 with CnLPAT in Camelina resulted in further accumulation of MCFAs in the seed (Table 2). However, co-expression of CnLPAT with the C8/C10-specific FatB ChFatB2 did not increase 8:0 or 10:0 fatty acid levels in Camelina seeds relative to expression of ChFatB2 alone (Table 2). Analysis of sn-2 fatty acids of TAG confirmed the effect of CnLPAT to increase MCFAs in TAG. Myristic acid (14:0) and lauric acid (12:0) were detected in the sn-2 position of TAG in the co-expression lines of CpFatB2 with CnLPAT and UcFatB1 with CnLPAT, respectively, while these fatty acids were 5- to 10-fold lower in the absence of CnLPAT (Fig. 5A, B). Interestingly, 10:0 was not detected at the sn-2 position of TAG in Camelina seeds co-expressing ChFatB2 and CnLPAT (Fig. 5C), indicating that CnLPAT has substrate specificity for CoA esters of 12:0 preferentially and 14:0 to a lesser extent, but is not active with 10:0-CoA (Table 2).


Toward production of jet fuel functionality in oilseeds: identification of FatB acyl-acyl carrier protein thioesterases and evaluation of combinatorial expression strategies in Camelina seeds.

Kim HJ, Silva JE, Vu HS, Mockaitis K, Nam JW, Cahoon EB - J. Exp. Bot. (2015)

Fatty acid composition of the sn-2 position of seed oil TAG. Fatty acid composition of the sn-2 position of seed oil TAG in plants expressing FatB genes alone or in combination with CnLPAT as determined by lipase digestion-based analyses. The data represent averages of four biological replicates ±SD. (A) CpFatB2 and CpFatB2 with CnLPAT. (B) UcFatB1 and UcFatB1 with CnLPAT. (C) ChFatB2 and ChFatB2 with CnLPAT.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License 1 - License 2
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Figure 5: Fatty acid composition of the sn-2 position of seed oil TAG. Fatty acid composition of the sn-2 position of seed oil TAG in plants expressing FatB genes alone or in combination with CnLPAT as determined by lipase digestion-based analyses. The data represent averages of four biological replicates ±SD. (A) CpFatB2 and CpFatB2 with CnLPAT. (B) UcFatB1 and UcFatB1 with CnLPAT. (C) ChFatB2 and ChFatB2 with CnLPAT.
Mentions: To increase MCFA levels further in Camelina seeds, FatB polypeptides were co-expressed with a coconut lysophosphatidic acid acyltransferase (CnLPAT) that has been shown to insert 12:0 at the sn-2 position of the TAG glycerol backbone (Table 1), leading to 12:0 accumulation in transgenic B. napus seeds (Knutzon et al., 1999). Co-expression of either CpFatB2 or UcFatB1 with CnLPAT in Camelina resulted in further accumulation of MCFAs in the seed (Table 2). However, co-expression of CnLPAT with the C8/C10-specific FatB ChFatB2 did not increase 8:0 or 10:0 fatty acid levels in Camelina seeds relative to expression of ChFatB2 alone (Table 2). Analysis of sn-2 fatty acids of TAG confirmed the effect of CnLPAT to increase MCFAs in TAG. Myristic acid (14:0) and lauric acid (12:0) were detected in the sn-2 position of TAG in the co-expression lines of CpFatB2 with CnLPAT and UcFatB1 with CnLPAT, respectively, while these fatty acids were 5- to 10-fold lower in the absence of CnLPAT (Fig. 5A, B). Interestingly, 10:0 was not detected at the sn-2 position of TAG in Camelina seeds co-expressing ChFatB2 and CnLPAT (Fig. 5C), indicating that CnLPAT has substrate specificity for CoA esters of 12:0 preferentially and 14:0 to a lesser extent, but is not active with 10:0-CoA (Table 2).

Bottom Line: Expression of CpuFatB3 and CvFatB1 resulted in Camelina oil with capric acid (10:0), and CpuFatB4 expression conferred myristic acid (14:0) production and increased 16:0.Increases in lauric acid (12:0) and 14:0, but not 10:0, in Camelina oil and at the sn-2 position of triacylglycerols resulted from inclusion of a coconut lysophosphatidic acid acyltransferase specialized for MCFAs.Camelina lines presented here provide platforms for additional metabolic engineering targeting fatty acid synthase and specialized acyltransferases for achieving oils with high levels of jet fuel-type fatty acids.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.

No MeSH data available.


Related in: MedlinePlus