Cloning and Characterization of a Constitutive Lysophosphatidic Acid Acyltransferase 2 (LPAT2) Gene from Tropaeolum majus L~!2009-07-22~!2009-09-25~!2010-03-12~!

Abstract

The cloning and characterization of a lyso-phosphatidic acid acyltransferase (LPAT2; EC 2.3.1.51) from Tropaeolum majus from a 20,000 EST collection is described. The 1358 bp TmLPAT2 gene encodes a 42.6 kD polypeptide; the primary sequence has a membrane bound O-acyltransferase, MBOAT, Box I motif, and Boxes II, III and IV typical of LPATs from various species. Unlike many LPAT2s, the gene was constitutively expressed in all tissues. The TmLPAT2 functionality was confirmed by expression in a yeast LPAT deletion (SLC1-) mutant. The TmLPAT2 could use a range of acyl-CoAs as acyl donor, including 22:1-CoA and 20:1-CoA and either 18:1-LPA or 22:1-LPA as acyl acceptor. This new LPAT2 could enable the production of Brassica seed oils with enhanced levels of very long-chain fatty acids.

Increase in lysophosphatidate acyltransferase activity in oilseed rape (Brassica napus) increases seed triacylglycerol content despite its low intrinsic flux control coefficient

“…) are increased in LPAAT‐OE. We do not believe that the changes found in DAG, PC and TAG were due to the substrate preference of the nasturtium enzyme per se , as this enzyme uses palmitoyl‐CoA and oleoyl‐CoA equally, as judged by specificity measurements (Taylor et al ., ) and the oilseed rape LPAAT was also shown to use these substrates at equivalent rates in a selectivity assay (Berneth & Frentzen, ).…”

“…The construct used to produce overexpressing lines of B. napus was derived from a nasturtium (Tropaeolum majus) LPAAT gene New Phytologist (Taylor et al, 2010) whose activity was confirmed by complementation of a LPAAT deletion (SLC1 À ) mutant and has been shown to utilise 16:0-CoA and 18:1-CoA with about equal efficiency (Taylor et al, 2010). The details of the construct are shown in Fig.…”

Brassica carinata – a new molecular farming platform for delivering bio‐industrial oil feedstocks: case studies of genetic modifications to improve very long‐chain fatty acid and oil content in seeds

“…We have generated a collection of 20 000 ESTs from a library of subtracted developing nasturtium embryo cDNAs, which we will continue to ‘mine’ to isolate key genes to effect trierucin synthesis in B. carinata. To this end, we recently cloned and characterized an LPAT2 from nasturtium ( TmLPAT2 ) that will catalyze the synthesis of dierucoyl PA in the Kennedy pathway 107. Current experiments underway will re‐transform the Crambe KCS + RNAi FAD2 dual transgene B. carinata line with the TmLPAT2 to try to produce B. carinata prototypes with 70% erucic acid or better.…”

A genome-wide analysis of the lysophosphatidate acyltransferase (LPAAT) gene family in cotton: organization, expression, sequence variation, and association with seed oil content and fiber quality

“…Also, the overexpression of LPAATs increased seed oil content, thus reinforcing the utility of LPAAT genes as valuable biotechnological tools. Consequently, LPAAT homologs have been cloned from poached eggplant ( Limnanthes douglasii ) [ 22 ], A. thaliana [ 23 ], rapeseed [ 24 ], coconut [ 19 ], corn ( Zea mays ) [ 25 ], Indian cress ( Tropaeolum majus ) [ 26 ], Java olives ( Sterculia foetida ) [ 27 ], and castor [ 20 ]. The overexpression of a LPAAT gene from the SLCl-1 mutant yeast stain in A. thaliana and B. napus increased the incorporation of long-chain fatty acids into the sn -2 position of TAGs, leading to an increase in seed oil content by 8–48% [ 28 ].…”