THE CLONING AND PREL IM INARY CHARACTERIZATION OF A C18 ∶0Δ9 DESATURASE GENE FROM MARINE M ICROALGAE PAVLOVA VI RID IS

YUAN Dong-Juan; ZHOU Ke-Yuan; KANG Jing-Xuan; JIANG Li-Ming

Volume 33 Issue 4

Aug. 2014

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ACTA HYDROBIOLOGICA SINICA > 2009 > 33(4): 732-739.

YUAN Dong-Juan, ZHOU Ke-Yuan, KANG Jing-Xuan, JIANG Li-Ming. THE CLONING AND PREL IM INARY CHARACTERIZATION OF A C18 ∶0Δ9 DESATURASE GENE FROM MARINE M ICROALGAE PAVLOVA VI RID IS[J]. ACTA HYDROBIOLOGICA SINICA, 2009, 33(4): 732-739.

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THE CLONING AND PREL IM INARY CHARACTERIZATION OF A C18 ∶0Δ9 DESATURASE GENE FROM MARINE M ICROALGAE PAVLOVA VI RID IS

Department of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang524023, P. R. China;
2. Department of Medicine, Massachusetts General Hospital, and the Harvard Medical School, Boston, MA02114, USA

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supported in part by grants from the Guangdong Technology Plan Program (Grant 2005B10401011)

Received Date: January 13, 2002
Rev Recd Date: April 21, 2009
Published Date: July 24, 2009

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Abstract

In higher plant, Δ9 desaturase introduces the first double bond into saturated fatty acids, resulting inthe corresponding monounsaturated fatty acids.We have cloned gene, designated as PvfadA, for C18∶0Δ9 desaturase catalytic activity from Pavlova viridis by RT-PCR, RNA ligase mediated RACE (RLM-RACE) and Overlap-PCR strategy. This desaturase, when expressed in Escherichia coli, desaturated stearic acid to yield oleic acid, but did not desaturate other fatty acids in E. coli. These results indicate that PvFadA is specific to stearic acid. The deduced amino acid sequences of PvFadA show that a putative (D /E) X2HX～100(D /E) X2 H metal binding motif, which specifically existed in acyl-ACP desaturase. Moreover, Amino acids of PvFadA are similar in part to those of acyl-ACP desaturases from higher plant, such as A rabidopsis thaliana, Oryza sativa and Glycine max, but not to those of acyl-lipid desaturases of Cyanobacteria, and acyl-CoA desaturases of higher plant. In addition, the 3D2model structure of PvFadA is composed of 11 helixes, moreover, α3, α4, α6 andα7 form a core 42helix bundle to regards as an active center in this enzyme, similar as acyl-ACP desaturase of Ricinus communis and M ycobacterium tuberculosis H37Rv.
- Stearoyl-ACP,
- Δ9 desaturase,
- RLM-RACE PCR,
- Pavlova viridis,
- GC-MS

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[1]	Cahoon E B, Ohlrogge J B. Metabolic evidence for the involvement of a [delta] 4-palmitoyl-acyl carrier protein desaturase in petroselinic acid synthesis in coriander endosperm and transgenic tobacco cells [J]. Plant Physiol.1994, 104 (3) : 827-837
[2]	TocherD R, LeaverM J, Hodgson P A. Recent advances in the biochemistry and molecular biology of fatty acyl desaturases [J]. Prog Lipid Res, 1998, 37: 73-117
[3]	MocheM, Shanklin J, GhoshalA, et al. Azide and acetate complexes plus two iron-depleted crystal structures of the diiron enzyme delta 9 stearoyl-acyl carrier protein desaturase1 Implications for oxygen activation and catalytic intermediates [J]. J B iol Chem., 2003, 278 (27) : 25072-25080
[4]	DyerD H, Lyle K S, Rayment I, et al. X-ray structure of putative acyl-ACP desaturase DesA2 from Mycobacterium tuberculosis H37Rv [J]. Protein Sci1, 2005, 14 (6) : 1508-1517
[5]	Sakamoto T, Wada H, Nishida I, et al.Delta 9 Acyl-lipid desaturases of cyanobacteria1Molecular cloning and substrate specificities in terms of fatty acids, snpositions, and polar head groups [J].J B iol Chem., 1994, 269 (41) : 25576-25580
[6]	Domergue F, Spiekermann P, Lerchl J, et al. New insight intoPhaeodactylum tricornutum fatty acid metabolism. Cloning and functional characterization of plastidial and microsomal delta122 fatty acid desaturases [J]. Plant Physiol. 2003, 131 (4) :1648-1660
[7]	Magnuson K, Jackowski S, Rock C O, et al. Regulation of fatty acid biosynthesis in Escherichia coli [J].M icrobiol Rev., 1993, 57 (3) : 522-542
[8]	Ohlrogge J, Browse J. Lipid biosynthesis [J]. Plant Cell,1995, 7 (7) : 957-970
[9]	Cahoon E B, Mills L A, Shanklin J1 Modification of the fatty acid composition of Escherichia coli by coexpression of a plant acyl-acyl carrier protein desaturase and ferredoxin [J]. Journal of Bacteriology, 1996, 178 (3) : 936-939
[10]	Serrano-Vega M J, Venegas-Calerón M, Garcēs R, et al. Cloning and expression of fatty acids biosynthesis key enzymes from sunflower (Helianthus annuus L.) in Escherichia coli [J].Journal of Chromatography B, 2003, 786 (1-2) : 221-228
[11]	Whittle E, Cahoon EB, Subrahmanyam S, et al.A multifunctional acyl-acyl carrier protein desaturase from Hedera helix L. (English Ivy) can synthesize 16-and 18-carbon monoene and diene products [J].J B iol Chem, 2005, 280 (31) : 28169-28176
[12]	KaterM M, Koningstein GM, Nijkamp H J, et al. The use of a hybrid genetic system to study the functional relationship between prokaryotic and plant multi-enzyme fatty acid synthetase complexes [J]. PlantM ol B iol., 1994, 25 (5) : 771-790
[13]	Sambrook J, Russell D W. Molecular Cloning: A Laboratory Manual, 3rd ed [M]. New York: Cold Spring Harbor Press1 2001
[14]	KangJ X, WangJ.A simplifiedmethod for analysisofpolyunsaturated fatty acids [J]. BMC B iochem, 2005, 6: 5
[15]	Thompson J D, Gibson TJ, Plewniak F, et al. The CLUSTAL_X windows interface: flexible strategies formultiple sequence alignment aided by quality analysis tools [J]. Nucleic Acids Res, 1997, 25: 4876-4882
[16]	Lindqvist Y, HuangW, Schneider G, et al. Crystal structure of delta 9 stearoyl-acyl carrier protein desaturase from castor seed and its relationship to other di2iron proteins [J]. EMBO J,1996, 15: 4081-4092
[17]	Bryson K, McGuffin L J, Marsden R L, et al. Protein structure prediction servers atUniversity College London [J]. Nucl. Acids Res, 2005, 33: W36-38
[18]	McGuffin L J, Bryson K, JonesD T. The PSIPRED protein structure prediction server [J].B ioinformatics, 2000, 16: 404-405
[19]	JonesD T. Protein secondary structure prediction based on position-specific scoring matrices [J]. J. M ol. B iol, 1999, 292:195-202
[20]	Jones D T. GenTHREADER: an efficient and reliable protein fold recognition method for genomic sequences [J]. J.M ol. B iol, 1999, 287: 797-815
[21]	McGuffin L J, Jones D T. Improvement of the GenTHREADER method for genomic fold recognition [J].B ioinformatics, 2003, 19: 874-881
[22]	Deleage G., Clerc F F, RouxB, et al. ANTHEPROT: a package for protein sequence analysis using a microcomputer [J].Comput Appl B iosci, 1988, 4 (3) : 351-363
[23]	Deleage G, Combet C, Blanchet C, et al. ANTHEPROT: an integrated protein sequence analysis softwarewith client/server capabilities [J]. Computers in B iology and Medicine, 2001, 31: 259-267
[24]	Shanklin J, Somerville C1 Stearoyl2Acyl2Carrier2Protein desaturase from higher plants is structurally unrelated to the animal and fungal homologs [J]. Proc.Natl.Acad. Sci.USA, 1991, 88: 2510-2514
[25]	FoxB G, Shanklin J, Somerville C, et al. Stearoyl-Acyl carrier proteinΔ9 desaturase from ricinus communis is a Diiron-Oxo protein [J]. Proc. Natl1 Acad. Sci. USA, 1993, 90: 2486-2490
[26]	Cahoon E B, Lindqvist Y, Schneider G, et al. Redesign of soluble fatty acid desaturases from plants for altered substrate specificity and double bond position [J].. Proc1 Natl1 Acad1 Sci. USA, 1997, 94: 4872-4877
[27]	EdwardW, Shanklin J. EngineeringΔ9216∶02Acyl Carrier Protein (ACP) desaturase specificity based on combinatorial saturation mutagenesis and logical redesign of the castorΔ9218∶02ACP desaturase [J]. J B iol Chem, 2001, 276: 21500-21505
[28]	Tong L, Shu2Ming P, Wu2Yuan D, et al. Characterization of a new stearoyl-acyl carrier protein desaturase gene from Jatropha curcas [J]. B iotechnology Letter, 2006, 28 (9) : 657-662
[29]	DyerD H, Lyle K S, Rayment I, et al. X-ray structure of pupative ayl-ACP desaturase DesA2 from Mycobacterium tuberculosis H37Rv [J]. Protein Sci., 2005, 14 (6) : 1508-1517

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THE CLONING AND PREL IM INARY CHARACTERIZATION OF A C18 ∶0Δ9 DESATURASE GENE FROM MARINE M ICROALGAE PAVLOVA VI RID IS

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