Acyl-lipid thioesterase1–4 from Arabidopsis thaliana form a novel family of fatty acyl–acyl carrier protein thioesterases with divergent expression patterns and substrate specificities

• Published in: Plant molecular biology Vol. 84; no. 4-5; pp. 549 - 563
• Main Authors: Pulsifer, Ian P, Lowe, Christine, Narayaran, Swara A, Busuttil, Alia S, Vishwanath, Sollapura J, Domergue, Frédéric, Rowland, Owen
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01.03.2014; Springer Netherlands; Springer Nature B.V
• Subjects: Acyl Carrier Protein - metabolism; Amino Acid Sequence; amino acid sequences; Amino acids; anthers; Arabidopsis; Arabidopsis - enzymology; Arabidopsis - genetics; Arabidopsis Proteins - genetics; Arabidopsis Proteins - metabolism; Arabidopsis thaliana; Biochemistry; Biomedical and Life Sciences; corolla; E coli; Escherichia coli; Fatty acids; Fatty Acids - metabolism; free fatty acids; gene expression; genes; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; hydrolysis; In Situ Hybridization; Isoenzymes - genetics; Isoenzymes - metabolism; Life Sciences; Liliopsida; Lipids; Metabolites; Microscopy, Confocal; Molecular biology; Molecular Sequence Data; Multigene Family; Mutation; oxidation; Plant biology; Plant Pathology; Plant Sciences; Plant tissues; Plants, Genetically Modified; plastids; Plastids - metabolism; Proteins; Reverse Transcriptase Polymerase Chain Reaction; Sequence Homology, Amino Acid; Substrate Specificity; Substrates; Thiolester Hydrolases - genetics; Thiolester Hydrolases - metabolism; Transcriptome; Thioesterase; Methylketone; Plastid; Fatty acid; Acyl carrier protein
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1007/s11103-013-0151-z

Hydrolysis of fatty acyl thioester bonds by thioesterases to produce free fatty acids is important for dictating the diversity of lipid metabolites produced in plants. We have characterized a four-member family of fatty acyl thioesterases from Arabidopsis thaliana, which we have called acyl-lipid thioesterase1 (ALT1), ALT2, ALT3, and ALT4. The ALTs belong to the Hotdog fold superfamily of thioesterases. ALT-like genes are present in diverse plant taxa, including dicots, monocots, lycophytes, and microalgae. The four Arabidopsis ALT genes were found to have distinct gene expression profiles with respect to each other. ALT1 was expressed specifically in stem epidermal cells and flower petals. ALT2 was expressed specifically in root endodermal and peridermal cells as well as in stem lateral organ boundary cells. ALT3 was ubiquitously expressed in aerial and root tissues and at much higher levels than the other ALTs. ALT4 expression was restricted to anthers. All four proteins were localized in plastids via an N-terminal targeting sequence of about 48 amino acids. When expressed in Escherichia coli, the ALT proteins used endogenous fatty acyl–acyl carrier protein substrates to generate fatty acids that varied in chain length (C6–C18), degree of saturation (saturated and monounsaturated), and oxidation state (fully reduced and β-ketofatty acids). Despite their high amino acid sequence identities, each enzyme produced a different profile of lipids in E. coli. The biological roles of these proteins are unknown, but they potentially generate volatile lipid metabolites that have previously not been reported in Arabidopsis.