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Identification of amino acids related to catalytic function of Sulfolobus solfataricus P1 carboxylesterase by site-directed mutagenesis and molecular modeling
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  • Journal title : BMB Reports
  • Volume 49, Issue 6,  2016, pp.349-354
  • Publisher : Korean Society for Biochemistry and Molecular Biology
  • DOI : 10.5483/BMBRep.2016.49.6.077
 Title & Authors
Identification of amino acids related to catalytic function of Sulfolobus solfataricus P1 carboxylesterase by site-directed mutagenesis and molecular modeling
Choi, Yun-Ho; Lee, Ye-Na; Park, Young-Jun; Yoon, Sung-Jin; Lee, Hee-Bong;
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 Abstract
The archaeon Sulfolobus solfataricus P1 carboxylesterase is a thermostable enzyme with a molecular mass of 33.5 kDa belonging to the mammalian hormone-sensitive lipase (HSL) family. In our previous study, we purified the enzyme and suggested the expected amino acids related to its catalysis by chemical modification and a sequence homology search. For further validating these amino acids in this study, we modified them using site-directed mutagenesis and examined the activity of the mutant enzymes using spectrophotometric analysis and then estimated by homology modeling and fluorescence analysis. As a result, it was identified that Ser151, Asp244, and His274 consist of a catalytic triad, and Gly80, Gly81, and Ala152 compose an oxyanion hole of the enzyme. In addition, it was also determined that the cysteine residues are located near the active site or at the positions inducing any conformational changes of the enzyme by their replacement with serine residues.
 Keywords
Carboxylesterase;Molecular modeling;Site-directed mutagenesis;Sulfolobus solfataricus P1;Thermostability;
 Language
English
 Cited by
 References
1.
Okuda H (1990) Esterases; in A study of enzymes, Kuby SA (eds.), 563-577, Vol. 2, CRC press, Boca Raton, FL, USA

2.
Jaeger KE, Dijkstra BW and Reetz MT (1999) Bacterial biocatalysts: molecular biology, threedimensional structures, and biotechnological applications of lipases. Annu Rev Microbiol 53, 315-351 crossref(new window)

3.
Arpigny JL and Jaeger KE (1999) Bacterial lipolytic enzymes: classification and properties. Biochem J 343, 177-183 crossref(new window)

4.
Rao L, Xue Y, Zhou C et al (2011) A thermostable esterase from Thermoanaerobacter tengcongensis opening up a new family of bacterial lipolytic enzymes. Biochim Biophys Acta 1814, 1695-1702 crossref(new window)

5.
Holm C, Osterlund T, Laurell H and Contreras JA (2000) Molecular mechanisms regulating hormone-sensitive lipase and lipolysis. Annu Rev Nutr 20, 365-393 crossref(new window)

6.
Holmquist M (2000) Alpha/beta-hydrolase fold enzymes: structures, functions and mechanisms. Curr Protein Pept Sci 1, 209-235 crossref(new window)

7.
Schrag JD and Cygler M (1997) Lipases and alpha/beta hydrolase fold. Methods Enzymol 284, 85-107 crossref(new window)

8.
De Simone G, Menchise V, Manco G et al (2001) The crystal structure of a hyperthermophilic carboxylesterase from the archaeon Archaeoglobus fulgidus. J Mol Biol 314, 507-518 crossref(new window)

9.
Byun JS, Rhee JK, Kim ND et al (2007) Crystal structure of hyperthermophilic esterase EstE1 and the relationship between its dimerization and thermostability properties. BMC Struct Biol 7:47, 1-11 crossref(new window)

10.
Palm GJ, Fernández-Álvaro E, Bogdanović X et al (2011) The crystal structure of an esterase from the hyperthermophilic microorganism Pyrobaculum calidifontis VA1 explains its enantioselectivity. Appl Microbiol Biotechnol 91, 1061-1072 crossref(new window)

11.
Angkawidjaja C, Koga Y, Takano K and Kanaya S (2012) Structure and stability of a thermostable carboxylesterase from the thermoacidophilic archaeon Sulfolobus tokodaii. FEBS J 279, 3071-3084 crossref(new window)

12.
Panda T and Gowrishankar BS (2005) Production and applications of esterases. Appl Microbiol Biotechnol 67, 160-169 crossref(new window)

13.
Brock TD, Brock KM, Belly RT and Weiss RL (1972) Sulfolobus: a new genus of sulfur oxidizing bacteria living at low pH and high temperature. Arch Microbiol 84, 54-68

14.
Park Y, Choi SY and Lee HB (2006) A carboxylesterase from the thermoacidophilic archaeon Sulfolobus solfataricus P1; purification, characterization, and expression. Biochim Biophys Acta 1760, 820-828 crossref(new window)

15.
Nam JK, Park YJ and Lee HB (2013) Cloning, expression, and characterization of a novel thermostable esterase from the archaeon Sulfolobus solfataricus P1. J Mol Catal B: Enzymatic 94, 95-103 crossref(new window)

16.
Park YJ, Yoon SJ and Lee HB (2008) A novel thermostable arylesterase from the archaeon Sulfolobus solfataricus P1: purification, characterization, and expression. J Bacteriol 190, 8086-8095 crossref(new window)

17.
Pathak D, Ashley G and Ollis D (1991) Thiol protease-like active site found in the enzyme dienelactone hydrolase: localization using biochemical, genetic, and structural tools. Proteins 9, 267-279 crossref(new window)

18.
Park YJ, Yoon SJ and Lee HB (2010) A novel dienelactone hydrolase from the thermoacidophilic archaeon Sulfolobus solfataricus P1: Purification, characterization, and expression. Biochim Biophys Acta 1800, 1164-1172 crossref(new window)

19.
Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, New York, USA

20.
Qin Y and Qu Y (2014) Asn124 of Cel5A from Hypocrea jecorina not only provides the N-glycosylation site but is also essential in maintaining enzymatic activity. BMB Rep 47, 256-261 crossref(new window)

21.
Lim CJ, Jeon JE, Jeong SK et al (2015) Growth hormonereleasing peptide-biotin conjugate stimulates myocytes differentiation through insulin-like growth factor-1 and collagen type I. BMB Rep 48, 501-506 crossref(new window)

22.
Roy A, Kucukural A and Zhang Y (2010) I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc 5, 725-738 crossref(new window)

23.
Zhang Y (2008) I-TASSER server for protein 3D structure prediction. BMC Bioinformatics 9:40