Biochemical characteristics of AtFAR2, a fatty acid reductase from Arabidopsis thaliana that reduces fatty acyl-CoA and -ACP substrates into fatty alcohols

• Authors: Thuy Doan , Anders Carlsson , Sten Stymne , Per Hofvander
• Languages of publication: EN

Abstracts

EN

Fatty alcohols and derivatives are important for proper deposition of a functional pollen wall. Mutations in specific genes encoding fatty acid reductases (FAR) responsible for fatty alcohol production cause abnormal development of pollen. A disrupted AtFAR2 (MS2) gene in Arabidopsis thaliana results in pollen developing an abnormal exine layer and a reduced fertility phenotype. AtFAR2 has been shown to be targeted to chloroplasts and in a purified form to be specific for acyl-ACP substrates. Here, we present data on the in vitro and in planta characterizations of AtFAR2 from A. thaliana and show that this enzyme has the ability to use both, C16:0-ACP and C16:0-CoA, as substrates to produce C16:0-alcohol. Our results further show that AtFAR2 is highly similar in properties and substrate specificity to AtFAR6 for which in vitro data has been published, and which is also a chloroplast localized enzyme. This suggests that although AtFAR2 is the major enzyme responsible for exine layer functionality, AtFAR6 might provide functional redundancy to AtFAR2.

Keywords

EN

Arabidopsis thaliana; chloroplast; fatty acyl-CoA/ACP reductase; fatty alcohol; fatty aldehyde

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2016
• Volume: 63
• Issue: 3
• Pages: 565-570

Dates

published

2016

received

2016-01-01

revised

2016-02-04

accepted

2016-02-16

(unknown)

2016-06-08

Contributors

author

Thuy Doan

• Department of Biology, Faculty of Science, Nong Lam University, Ho Chi Minh City, Vietnam

author

Anders Carlsson

• Department of Plant Breeding, Swedish University of Agricultural Sciences, P.O. Box 101, 230 53 Alnarp, Sweden

author

Sten Stymne

• Department of Plant Breeding, Swedish University of Agricultural Sciences, P.O. Box 101, 230 53 Alnarp, Sweden

author

Per Hofvander

• Department of Plant Breeding, Swedish University of Agricultural Sciences, P.O. Box 101, 230 53 Alnarp, Sweden

References

• Aarts MG, Hodge R, Kalantidis K, Florack D, Wilson ZA, Mulligan BJ, Stiekema WJ, Scott R, Pereira A (1997) The Arabidopsis MALE STERILITY 2 protein shares similarity with reductases in elongation/condensation complexes. The Plant Journal 12: 615-623. doi: 10.1046/j.1365-313X.1997.00615.x.
• Bligh E, Dyer W (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911-917. doi: 10.1139/o59-099.
• Chacon MG, Fournier AE, Tran F, Dittrich-Domergue F, Pulsifer IP, Domergue F, Rowland O (2013) Identification of amino acids conferring chain length substrate specificities on fatty alcohol-forming reductases FAR5 and FAR8 from Arabidopsis thaliana. J Biol Chem 288: 30345-30355. doi: 10.1074/jbc.M113.499715.
• Chen W, Yu XH, Zhang K, Shi J, Schreiber L, Shanklin J, Zhang D (2011) Male Sterile2 encodes a plastid-localized fatty acyl carrier protein reductase required for pollen exine development in Arabidopsis thaliana. Plant Physiol 157: 842-853. doi: 10.1104/pp.111.181693.
• Doan TT, Carlsson AS, Hamberg M, Bülow L, Stymne S, Olsson P (2009) Functional expression of five Arabidopsis fatty acyl-CoA reductase genes in Escherichia coli. J Plant Physiol 166: 787-796. doi: 10.1016/j.jplph.2008.10.003.
• Doan TT, Domergue F, Fournier AE, Vishwanath SJ, Rowland O, Moreau P, Wood CC, Carlsson AS, Hamberg M, Hofvander P (2012) Biochemical characterization of a chloroplast localized fatty acid reductase from Arabidopsis thaliana. Biochim Biophys Acta 1821: 1244-1255. doi: 10.1016/j.bbalip.2011.10.019.
• Dobritsa AA, Shrestha J, Morant M, Pinot F, Matsuno M, Swanson R, Moller BL, Preuss D (2009) CYP704B1 is a long-chain fatty acid {omega}-hydroxylase essential for sporopollenin synthesis in pollen of Arabidopsis. Plant Physiol 151: 574-589. doi: 10.1104/pp.109.144469.
• Domergue F, Vishwanath SJ, Joubes J, Ono J, Lee JA, Bourdon M, Alhattab R, Lowe C, Pascal S, Lessire R, Rowland O (2010) Three Arabidopsis fatty acyl-coenzyme A reductases FAR1 FAR4 and FAR5 generate primary fatty alcohols associated with suberin deposition. Plant Physiol 153: 1539-1554. doi: 10.1104/pp.110.158238.
• Kolattukudy PE (1970) Reduction of fatty acids to alcohols by cell-free preparations of Euglena gracilis. Biochemistry 9: 1095-1102. doi: 10.1021/bi00807a007.
• Koncz C, Schell J (1986) The promoter of T L-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium binary vector. Mol General Genet MGG 204: 383-396. doi: 10.1007/BF00331014.
• Lueking DR, Goldfine H (1975) The involvement of guanosine 5-diphosphate-3-diphosphate in the regulation of phospholipid biosynthesis in Escherichia coli. Lack of ppGpp inhibition of acyltransfer from acyl-ACP to sn-glycerol 3-phosphate. J Biol Chem 250: 4911. doi: 250/13/4911.long.
• Metz JG, Pollard MR, Anderson L, Hayes TR, Lassner MW (2000) Purification of a jojoba embryo fatty acyl-coenzyme A reductase and expression of its cDNA in high erucic acid rapeseed. Plant Physiol 122: 635-644. doi: 10.1104/pp.122.3.635.
• Nallamsetty S, Austin BP, Penrose KJ, Waugh DS (2005) Gateway vectors for the production of combinatorially-tagged His6-MBP fusion proteins in the cytoplasm and periplasm of Escherichia coli. Protein Sci 14: 2964-2971.
• Pollard M, Beisson F, Li Y, Ohlrogge JB (2008) Building lipid barriers: biosynthesis of cutin and suberin. Trends Plant Sci 13: 236-246. doi: 10.1016/j.tplants.2008.03.003.
• Post-Beittenmiller D (1996) Biochemistry and molecular biology of wax production in plants. Annu Rev Plant Physiol Plant Mol Biol 47: 405-430. doi: 10.1146/annurev.arplant.47.1.405.
• Rowland O, Domergue F (2012) Plant fatty acyl reductases: Enzymes generating fatty alcohols for protective layers with potential for industrial applications. Plant Sci 193-194: 28-38. doi: 10.1016/j.plantsci.2012.05.002.
• Rowland O, Zheng H, Hepworth SR, Lam P, Jetter R, Kunst L (2006) CER4 encodes an alcohol-forming fatty acyl-coenzyme A reductase involved in cuticular wax production in Arabidopsis. Plant Physiol 142: 866-877. doi: 10.1104/pp.106.086785.
• Shi J, Tan H, Yu XH, Liu Y, Liang W, Ranathunge K, Franke RB, Schreiber L, Wang Y, Kai G, Shanklin J, Ma H, Zhang D (2011) Defective pollen wall is required for anther and microspore development in rice and encodes a fatty acyl carrier protein reductase. The Plant Cell 23: 2225-2246. doi: 10.1105/tpc.111.087528.
• Vioque J, Kolattukudy PE (1997) Resolution and purification of an aldehyde-generating and an alcohol-generating fatty acyl-CoA reductase from pea leaves (Pisum sativum L.). Arch Biochem Biophys 340: 64-72. doi: 10.1006/abbi.1997.9932.
• Wang A, Xia Q, Xie W, Dumonceaux T, Zou J, Datla R, Selvaraj G (2002) Male gametophyte development in bread wheat (Triticum aestivum L.): molecular cellular and biochemical analyses of a sporophytic contribution to pollen wall ontogeny. The Plant Journal 30: 613-623. doi: 10.1046/j.1365-313X.2002.01313.x.
• Wood CC, Petrie JR, Shrestha P, Mansour MP, Nichols PD, Green AG, Singh SP (2009) A leaf-based assay using interchangeable design principles to rapidly assemble multistep recombinant pathways. Plant Biotechnol J 7: 914-924. doi: 10.1111/j.1467-7652.2009.00453.x.

Identifiers

DOI

10.18388/abp.2016_1245