Cloning and Bioinformatics Analysis of VdAS1 Gene in Vaccinium dunalianum (Ericaceae)

doi:10.7677/ynzwyj201514057

Abstract

Abstract:

Arbutin was widely used in cosmetics as a whitening agent because of blocking the synthesis of melanin as the inhibitor of tyrosinase. Arbutin synthase (AS) as one member of UDP glycosyltransferases (UGTs) which belongs to the glycosyltransferase (GTs) family is a ratelimiting enzyme in arbutin synthesis from hydroquinone. To reveal the biological function and provide scientific basis of application on plant genetic engineering for Vaccinium dunalianum arbutin synthase (VdAS), a gene VdAS1 encoding VdAS1 was isolated by RTPCR combined with RACEPCR from Vdunalianum. The fulllength cDNA clone comprised 1577 nucleotides consisting of a 17nucleotide 5′untranslated region, an open reading frame of 1428 nucleotides, and a 132nucleotide 3′untranslated region. The open reading frame encoded a putative VdAS1 comprising 475 amino acid residues with molecular weight of 5222kD. and isoelectric point of 574. The total numbers of negative charged residues (Asp+Glu) and positive charged residues (Arg+Lys) in VdAS1 were 53 and 45, respectively. VdAS1 shares 74% similarity with UGT in wolfberry (Lycium barbarum). It was predictive VdAS1 had no signal peptide and transmembrane structure. The main parts of predicted secondary structures of VdAS1 were ahelices and random coils. The study results established the foundation for the heterologous expression and functional analysis of VdAS1.

Key words: Vaccinium dunalianum, Arbutin synthase, Gene cloning, Bioinformatics analysis

CLC Number:

Q 75

DU Wei, DING Yong, ZHU Dong-Yang, YIN Ji-Ting, LIU Xiao-Zhu, ZHAO Ping. Cloning and Bioinformatics Analysis of VdAS1 Gene in Vaccinium dunalianum (Ericaceae)[J]. Plant Diversity, 2015, 37(01): 71-77.

References

Akiu S, Suzuki Y, Asahara T et al., 1991. Inhibitory effect of arbutin on melanogenesisbiochemical study using cultured B16 melanoma cells［J］. Nihon Hifuka Gakkai Zasshi, 101 (6): 609—613
Arend J, Warzecha H, Stckigt J, 2000. Hydroquinone: Oglucosyltransferase from cultivated Rauvolfia cells: enrichment and partial amino acid sequences［J］. Phytochemistry, 53 (2): 187—193
Arend J, Warzecha H, Hefner T et al., 2001. Utilizing genetically engineered bacteria to produce plantspecific glucosides［J］. Biotechnology and Bioengineering, 76 (2): 126—131
Barvkar VT, Pardeshi VC, Kale SM et al., 2012. Phylogenomic analysis of UDP glycosyltransferase1 multigene family in Linum usitatissimum identified genes with varied expression patterns［J］. BMC Genomics, 13: 175—188
Baumann L, WooleryLloyd H, Friedman A, 2009. “Natural” ingredientsin cosmetic dermatology［J］. Journal of Drugs in Dermatology, 8 (6 suppl): s5—9
Breton C, FournelGigleux SF, Palcic MM, 2012. Recent structures, evolution and mechanisms of glycosyltransferases［J］. Current Opinion in Structural Biology, 22 (5): 540—549
Cao PJ, Bartley LE, Jung KH et al., 2008. Construction of a rice glycosyltransferase phylogenomic database and identication of ricediverged glycosyltransferases［J］. Molecular Plant, 1 (5): 858—877
Hefner T, Arend J, Warzecha H et al., 2002. Arbutin synthase, a novel member of the NRD1β glycosyltransferase family, is a unique multifunctional enzyme converting various natural products and xenobiotics［J］. Bioorganic & Medicinal Chemistry, 10 (6): 1731—1741
Hefner T, Stckigt J, 2003. Probing suggested catalytic domains of glycosyltransferases by sitedirected mutagenesis［J］. European Journal of Biochemistry, 270 (3): 533—538
Kapitonov D, Yu RK, 1999. Conserved domains of glycosyltransferases［J］. Glycobiology, 9 (10): 961—978
Kumar S, Tamura K, Nei M, 1994. MEGA: Molecular evolutionary genetics analysis software for microcomputers［J］. Computer Applications in the Biosciences, 10 (2): 189—191
Li Y, Baldauf S, Lim EK et al., 2001. Phylogenetic analysis of the UDPglycosyltransferase multigene family of Arabidopsis thaliana［J］. Journal of Biological Chemistry, 276 (6): 4338—4343
Lutterbach R, Stckigt J, 1992. Highyield formation of arbutin from hydroquinone by cellsuspension cultures of Rauwolfia serpentina［J］. Helvetica Chimica Acta, 75 (6): 2009—2011
Maeda K, Fukuda M, 1996. Arbutin: mechanism of its depigmenting action in human melanocyte culture［J］. Jouranl Pharmacology and Experiment Therapeutics, 276 (2): 765—769Paquette S, Moller BL, Bak S, 2003. On the origin of family 1 plant glycosyltransferases［J］. Phytochemistry, 62 (3): 399—413
Wang YF (王亚芳), Zhou YH (周宇辉), Zhang JJ (张建军), 2003. Pharmacodynamics of arbutin on relieving cough, dispelling phlegm and preventing asthma［J］. Chinese Traditional and Herbal Drugs (中草药), 34 (8): 739—741
Wei Y, Zhang ZJ, Zhang YT et al., 2007. Simple LC method chemiluminescence detection for simultaneous determination of arbutin and Lascorbic acid in whitening cosmetics［J］. Chromatographia, 65 (7): 443—446
Wu FR (吴方睿), 2013. Study on the expression, purification, crystallization, characterization of complex threedimensional structure and substrate extension of two important enzymes from Rauvolfia serpentina［D］. Hangzhou : Zhejiang University, 1—174
YonekuraSakakibara K, Hanada K, 2011. An evolutionary view of functional diversity in family 1 glycosyltransferases［J］. The Plant Journal, 66 (1): 182—193
Zhao P, Tanaka T, Hirabayashi K et al., 2008. Caffeoyl arbutin and related compounds from the buds of Vaccinium dunalianum［J］. Phytochemistry, 69 (18): 3087—3094
Zhu DY (朱东阳), Yin JT (尹继庭), Ding Y (丁勇) et al., 2013. The comparison on total RNA isolation from Vaccinium dunalianum leaf buds［J］. Journal of West China Forestry Science (西部林业科学), 42 (1): 81—85

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