全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

Enzymatic Properties and Mutational Studies of Chalcone Synthase from Physcomitrella patens

DOI: 10.3390/ijms13089673

Keywords: chalcone synthase, site-directed mutagenesis, active site, by-products

Full-Text   Cite this paper   Add to My Lib

Abstract:

PpCHS is a member of the type III polyketide synthase family and catalyses the synthesis of the flavonoid precursor naringenin chalcone from p-coumaroyl-CoA. Recent research reports the production of pyrone derivatives using either hexanoyl-CoA or butyryl-CoA as starter molecule. The Cys-His-Asn catalytic triad found in other plant chalcone synthase predicted polypeptides is conserved in PpCHS. Site directed mutagenesis involving these amino acids residing in the active-site cavity revealed that the cavity volume of the active-site plays a significant role in the selection of starter molecules as well as product formation. Substitutions of Cys 170 with Arg and Ser amino acids decreased the ability of the PpCHS to utilize hexanoyl-CoA as a starter molecule, which directly effected the production of pyrone derivatives (products). These substitutions are believed to have a restricted number of elongations of the growing polypeptide chain due to the smaller cavity volume of the mutant’s active site.

References

[1]  Jiang, C.; Schrommer, C.K.; Kim, S.Y.; Suh, D.Y. Cloning and characterization of chalcone synthase fron the moss, Physcomitrella patens. Phytochemistry 2006, 67, 2531–2540.
[2]  Harashima, S.; Takano, H.; Ono, K.; Takio, S. Chalcone synthase-like gene in liverwort, Marchantia palacea var diptera. Plant Cell Reprod 2004, 23, 167–173.
[3]  Pang, Y.; Shen, G.; Wu, W.; Liu, X.; Lin, J.; Tan, F.; Sun, X.; Tang, K. Characterization and expression of chalcone syntahse gene from Gikgo biloba. Plant Sci 2005, 168, 1525–1531.
[4]  Schaefer, D.G.; Zryd, J.-P. The moss Physcomitrella patens, Now and Then. Plant Physiol 2001, 127, 1430–1438.
[5]  Wolf, L.; Rizzini, L.; Stracke, R.; Ulm, R.; Rensing, S.A. The molecular and physiological responses of Physcomitrella patens to Ultraviolet-B-radiation. Plant Physiol 2010, 153, 1123–1134.
[6]  Koduri, P.K.; Gordon, G.S.; Barker, E.I.; Colpitts, C.C.; Ashton, N.W.; Suh, D.Y. Genome-wide analysis of the chalcone synthase superfamily genes of Physcomitrella patens. Plant Mol. Biol 2009, 72, 247–263.
[7]  Jez, J.M.; Noel, J.P. Mechanism of Chalcone Synthase. pKa of the catalytic cysteine and the role of the conserved histidine in a plant polyketide synthase. J. Biol. Chem 2000, 275, 39640–39646.
[8]  Morita, H.; Noguchi, H.; Schroder, J.; Abe, I. Novel polyketide synthesized with a higher plant stilbene synthase. European Journal of Biochemistry 2001, 268, 3759–3766.
[9]  Ferrer, J.L.; Bowman, M.E.; Dixon, R.A.; Noel, J.P. Structure of chalcone synthase and the molecular basis of plant polyketide biosynthesis. Nat. Struct. Biol 1999, 6, 775–784.
[10]  Zakaria, I.I.; Rahman, R.N.Z.; Salleh, A.B.; Mahiran, B. Bacteriocin-release protein mediated secretory expression of recombinant chalcone synthase in Escherichia coli. Appl. Biochem. Biotechnol 2011, 165, 737–747.
[11]  Zuurbier, K.W.M.; Leser, J.; Berger, T.; Hofte, A.J.P.; Schroder, G.; Verpoorte, R.; Schroder, J. 4-hyroxy-2-pyrone formation by chalcone synthase and stilbene synthase with nonphysiological substrates. Phytochemisty 1998, 49, 1945–1951.
[12]  Schuz, R.; Heller, W.; Hahlbrock, K. Substrate specificity of chalcone synthase from Petroselinum hortense. J. Biol. Chem 1983, 258, 6730–6734.
[13]  Funa, N.; Ohnishi, Y.; Ebizuka, Y.; Horinouchi, S. Alteration of reaction and substrate specificity of a bacterial type III polyketide synthase by site-directed mutagenesis. Biochem. J 2002, 367, 781–789.
[14]  Raharjo, T.J.; Chang, W.T.; Choi, Y.H.; Peltenburg-Looman, A.M.G.; Verpoorte, R. Olivetol as product of a polyketide synthase in Cannabis Sativa L. Plant Sci 2004, 166, 381–385.
[15]  Suh, D.-Y.; Kagami, J.; Fukuma, K.; Sankawa, U. Evidence for catalytic Cysteine-Histidine dyad in chalcone synthase. Biochem. Biophys. Res. Commun 2000, 275, 725–730.
[16]  Suh, D.-Y.; Fukuma, K.; Kagami, J.; Yamazaki, Y.; Shibuya, M.; Ebizuka, Y.; Sankawa, U. Identification of amino acid residues important in the cyclization reactions of chalcone and stilbene synthases. Biochem. J 2000, 350, 229–235.
[17]  Mallika, V.; Sivakumar, K.C.; Soniya, E.V. Implifications and physicochemical analyses of selected proteins of type III polyketide synthase family. Evol. Bioinforma 2011, 7, 41–53.
[18]  Lanz, T.; Tropf, S.; Marner, F.J.; Schroder, J.; Schroder, G. The role of cysteines in polyketide synthases. Site-directed mutagenesis of resveratrol and chalcone synthases, two key enzymes in different plant-specific pathways. J. Biol. Chem 1991, 266, 9971–9976.
[19]  Fukuma, K.; Neuls, E.D.; Ryberg, J.M.; Suh, D.-Y.; Sankawa, U. Mutational analysis of conserved outer sphere arginine residue of chalcone synthase. J. Biochem 2007, 142, 731–739.
[20]  MODELER Accelrys, San Diego, Available online: http://www.accelrys.com , accessed on 1 May 2012.
[21]  Sali, A.; Blundell, TL. Comparative protein modeling by satisfaction of spatial restraints. J. Mol. Biol. 1993, 234, 779–815.
[22]  , version 3.5.4; European Bioinformatics Institute: Heidelberg, Germany, 1994. Available online: http://www.biochem.ucl.ac.uk/~roman/procheck.html , accesed on 21 September 2011.
[23]  Errat, version 2.0; UCLA-DOE: Los Angeles, CA, USA, 2012. Available online: http://www.doe-mbi.ucla.edu/errat_server.html , accessed on 21 September 2011.
[24]  Zheng, Z.; Zuo, Z.; Liu, Z.; Tsai, K.; Liu, A.; Zou, G. Construction of a 3D model nattokinase, a novel fibrinolytic enzyme from Bacillus natto. A novel nucleophilic catalytic mechanism for nattokinase. J. Mol. Graphics Modell. 2005 2005, 23, 373–380.
[25]  Guerois, R.; Nielsen, J.E.; Serrano, L. Predicting changes in the stability of proteins and protein complexes: A study of more than 1000 mutations. J. Mol. Biol 2002, 320, 369–87.
[26]  Colovos, C.; Yeates, T.O. Verification of protein structures: Patterns of non-bonded atomic interaction. Protein Sci 1992, 2, 1511–1519.
[27]  YASARA, Biosciences GmbH, Available online: http://www.yasara.org , accessed on 1 May 2012.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133