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Vassilis Bouriotis

Vassilis Bouriotis

IMBB Group Leader (Full Professor, Univ. of Crete)
Email address
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Phone
+30-2810-394375
Fax
+30-2810-394055
Lab web page

Research in my laboratory is focused on characterization and applications of novel biocatalysts. Furthermore, using protein engineering (rational design and directed evolution) we aim to develop enzymes with improved properties for either established applications or areas of applications where enzymes have not previously been used.

Recent research in my laboratory centers on:

  •  P eptidoglycan N -acetyl g lucosamine deacetylases and O - a cetyl t ransferases from B.cereus and B.anthracis : These enzymes de- N -acetylate and O -acetylate respectively cell wall peptidoglycan to evade the lytic action of glycoside hydrolases secreted by the human immune system. We employ a combined biochemical crystallographic and genetic (knock out) analysis for these studies. These enzymes may serve as potential targets for the development of a new class of antibiotics specific for important human pathogenic bacteria.
  •  Chitin deacetylases from fungi: My group has a long tradition in the study of these enzymes. We have recently focused on the study of chitin deacetylases from the human pathogen Cryptococcus neoformans .
  •  Characterization of LmbE protein family enzymes from B. cereus.
  •  Protein engineering of cold adapted enzymes aiming to enhance their activity and/or thermal stability.

Specific applications and patents have emerged from our studies on chitin deacetylases and cold adapted enzymes.

A. Deli, D. Koutsioulis, P. Spiliotopoulou, K. Mavromatis, V. Fadouloglou, M. Kokkinidis and V. Bouriotis (2010) LmbE proteins from Bacillus cereus are de-N-acetylases with broad substrate specificity and highly conserved proteins in Bacillus anthracis. FEBS Journal 277: 2740-2753.

D. Koutsioulis, E. Wang, M. Tzanodaskalaki, D. Nikiforaki, A. Deli, G. Feller, P. Heikinheimo, and V. Bouriotis (2008) Directed evolution on the cold adapted properties of TAB5 alkaline phosphatase Protein Engineering. Design&Selection 21: 319-327.

I. G. Boneca, O. Dussurget, D. Cabanesc, M-A. Nahoric, S. Sousa, M. Lecuit, E. Psylinakis, V. Bouriotis, J-P. Hugoti, M. Giovanninik, A. Coylem, J. Bertinm, A. Namanep, J-C. Rousselle, N. Cayet, M-C. Prevost, V. Balloy, M. Chignard, D. J. Philpott, P. Cossart, and S. E. Girardin (2007) A critical role for peptidoglycan N-deacetylation in Listeria evasion from the host innate immune system PNAS 104: 997-1002.

E. Wang , D. Koutsioulis, H.-K. S. Leiros, O. A. Andersen, V. Bouriotis, E. Hough and P. Heikinheimo (2007) Crystal Structure of Alkaline Phosphatase from the Antarctic Bacterium TAB5 J. Mol. Biol. 366: 1318-1331.

E. Psylinakis, I.G. Boneca, K. Mavromatis, A. Deli, E. Hayhurst, S. J. Foster, K. M. Varum and V. Bouriotis (2005) Peptidoglycan N- Acetylglucosamine Deacetylases from Bacillus cereus, Highly Conserved Proteins in Bacillus anthracis. J.Biol.Chemistry 280: 30856-30863.

Publications from 2000 – today

• Unusual α-Carbon Hydroxylation of Proline Promotes Active-Site Maturation. 
Fadouloglou VE, Balomenou S, Aivaliotis M, Kotsifaki D, Arnaouteli S, Tomatsidou A, Efstathiou G, Kountourakis N, Miliara S, Griniezaki M, Tsalafouta A, Pergantis SA, Boneca IG, Glykos NM, Bouriotis V, Kokkinidis M.
J Am Chem Soc. 2017 Apr 19;139(15):5330-5337. doi: 10.1021/jacs.6b12209.

• Two Putative Polysaccharide Deacetylases Are Required for Osmotic Stability and Cell Shape Maintenance in Bacillus anthracis.
Arnaouteli S, Giastas P, Andreou A, Tzanodaskalaki M, Aldridge C, Tzartos SJ, Vollmer W, Eliopoulos E, Bouriotis V.
J Biol Chem. 2015 May 22;290(21):13465-78. doi: 10.1074/jbc.M115.640029.

• N-acetylglucosamine deacetylases modulate the anchoring of the gamma-glutamyl capsule to the cell wall of Bacillus anthracis.
Candela T, Balomenou S, Aucher W, Bouriotis V, Simore JP, Fouet A, Boneca IG.
Microb Drug Resist. 2014 Jun;20(3):222-30. doi: 10.1089/mdr.2014.0063.

• Structure determination of BA0150, a putative polysaccharide deacetylase from Bacillus anthracis.
Strunk RJ, Piemonte KM, Petersen NM, Koutsioulis D, Bouriotis V, Perry K, Cole KE.
Acta Crystallogr F Struct Biol Commun. 2014 Feb;70(Pt 2):156-9. doi: 10.1107/S2053230X13034262.

• Structure determination through homology modelling and torsion-angle simulated annealing: application to a polysaccharide deacetylase from Bacillus cereus.
Fadouloglou VE, Kapanidou M, Agiomirgianaki A, Arnaouteli S, Bouriotis V, Glykos NM, Kokkinidis M.
Acta Crystallogr D Biol Crystallogr. 2013 Feb;69(Pt 2):276-83. doi: 10.1107/S0907444912045829.

• Distinct functions of polysaccharide deacetylases in cell shape, neutral polysaccharide synthesis and virulence of Bacillus anthracis.
Balomenou S, Fouet A, Tzanodaskalaki M, Couture-Tosi E, Bouriotis V, Boneca IG
Mol Microbiol. 2013 Feb;87(4):867-83. doi: 10.1111/mmi.12137.

•  LmbE proteins from Bacillus cereus are de- N -acetylases with broad substrate specificity and are highly similar to proteins in Bacillus anthracis.
Deli A, Koutsioulis D, Fadouloglou VE, Spiliotopoulou P, Balomenou S, Arnaouteli S, Tzanodaskalaki M, Mavromatis K, Kokkinidis M, Bouriotis V.
FEBS J . 2010 Jul;277(13):2740-53.

•  Coordination sphere of the third metal site is essential to the activity and metal selectivity of alkaline phosphatases.
Koutsioulis D, Lyskowski A, Mäki S, Guthrie E, Feller G, Bouriotis V, Heikinheimo P.
Protein Sci . 2010 Jan;19(1):75-84.

•  Directed evolution on the cold adapted properties of TAB5 alkaline phosphatase.
Koutsioulis D, Wang E, Tzanodaskalaki M, Nikiforaki D, Deli A, Feller G, Heikinheimo P, Bouriotis V.
Protein Eng Des Sel . 2008 May;21(5):319-27.

•  Purification, crystallization and preliminary X-ray analysis of the peptidoglycan N-acetylglucosamine deacetylase BC1960 from Bacillus cereus in the presence of its substrate (GlcNAc)6.
Tsalafouta A, Psylinakis E, Kapetaniou EG, Kotsifaki D, Deli A, Roidis A, Bouriotis V, Kokkinidis M.
Acta Crystallogr Sect F Struct Biol Cryst Commun . 2008 Mar 1;64(Pt 3):203-5.

•  Crystal structure of the BcZBP, a zinc-binding protein from Bacillus cereus .
Fadouloglou VE, Deli A, Glykos NM, Psylinakis E, Bouriotis V, Kokkinidis M.
FEBS J . 2007 Jun;274(12):3044-54. Epub 2007 May 14.

•  A critical role for peptidoglycan N-deacetylation in Listeria evasion from the host innate immune system.
Boneca IG, Dussurget O, Cabanes D, Nahori MA, Sousa S, Lecuit M, Psylinakis E, Bouriotis V, Hugot JP, Giovannini M, Coyle A, Bertin J, Namane A, Rousselle JC, Cayet N, Prévost MC, Balloy V, Chignard M, Philpott DJ, Cossart P, Girardin SE.
Proc Natl Acad Sci U S A . 2007 Jan 16;104(3):997-1002.

•  Crystal structure of alkaline phosphatase from the Antarctic bacterium TAB5.
Wang E, Koutsioulis D, Leiros HK, Andersen OA, Bouriotis V, Hough E, Heikinheimo P.
J Mol Biol . 2007 Mar 2;366(4):1318-31. Epub 2006 Dec 2.

•  Purification, crystallization and preliminary X-ray analysis of the BseCI DNA methyltransferase from Bacillus stearothermophilus in complex with its cognate DNA.
Kapetaniou EG, Kotsifaki D, Providaki M, Rina M, Bouriotis V, Kokkinidis M.
Acta Crystallogr Sect F Struct Biol Cryst Commun . 2007 Jan 1;63(Pt 1):12-4.

•  Purification, crystallization and preliminary characterization of a putative LmbE-like deacetylase from Bacillus cereus .
Fadouloglou VE, Kotsifaki D, Gazi AD, Fellas G, Meramveliotaki C, Deli A, Psylinakis E, Bouriotis V, Kokkinidis M.
Acta Crystallogr Sect F Struct Biol Cryst Commun . 2006 Mar 1;62(Pt 3):261-4.

•  Crystallization and preliminary X-ray diffraction studies of an alcohol dehydrogenase from the Antarctic psychrophile Moraxella sp. TAE123.
Papanikolau Y, Tsigos I, Papadovasilaki M, Bouriotis V, Petratos K.
Acta Crystallogr Sect F Struct Biol Cryst Commun . 2005 Feb 1;61(Pt 2):246-8.

•  Peptidoglycan N-acetylglucosamine deacetylases from Bacillus cereus , highly conserved proteins in Bacillus anthracis .
Psylinakis E, Boneca IG, Mavromatis K, Deli A, Hayhurst E, Foster SJ, Vårum KM, Bouriotis V.
J Biol Chem . 2005 Sep 2;280(35):30856-63.

•  Evidence for increased local flexibility in psychrophilic alcohol dehydrogenase relative to its thermophilic homologue.
Liang ZX, Tsigos I, Lee T, Bouriotis V, Resing KA, Ahn NG, Klinman JP.
Biochemistry . 2004 Nov 23;43(46):14676-83.

•  Impact of protein flexibility on hydride-transfer parameters in thermophilic and psychrophilic alcohol dehydrogenases.
Liang ZX, Tsigos I, Bouriotis V, Klinman JP.
J Am Chem Soc . 2004 Aug 11;126(31):9500-1. Erratum in: J Am Chem Soc. 2006 Mar 15;128(10):3467.

•  Cold adaptation of a psychrophilic chitinase: a mutagenesis study.
Mavromatis K, Feller G, Kokkinidis M, Bouriotis V.
Protein Eng . 2003 Jul;16(7):497-503.

•  Mode of action and antifungal properties of two cold-adapted chitinases.
Mavromatis K, Lorito M, Woo SL, Bouriotis V.
Extremophiles. 2003 Oct;7(5):385-90.

•  Carbohydrate esterase family 4 enzymes: substrate specificity.
Caufrier F, Martinou A, Dupont C, Bouriotis V.
Carbohydr Res . 2003 Mar 28;338(7):687-92.

•  Exploring the role of a glycine cluster in cold adaptation of an alkaline phosphatase.
Mavromatis K, Tsigos I, Tzanodaskalaki M, Kokkinidis M, Bouriotis V.
Eur J Biochem . 2002 May;269(9):2330-5.

•  Expression, purification, and characterization of a cobalt-activated chitin deacetylase (Cda2p) from Saccharomyces cerevisiae .
Martinou A, Koutsioulis D, Bouriotis V.
Protein Expr Purif . 2002 Feb;24(1):111-6.

•  Engineering the properties of a cold active enzyme through rational redesign of the active site.
Tsigos I, Mavromatis K, Tzanodaskalaki M, Pozidis C, Kokkinidis M, Bouriotis V.
Eur J Biochem . 2001 Oct;268(19):5074-80.

•  Modular structure, local flexibility and cold-activity of a novel chitobiase from a psychrophilic Antarctic bacterium.
Lonhienne T, Zoidakis J, Vorgias CE, Feller G, Gerday C, Bouriotis V.
J Mol Biol . 2001 Jul 6;310(2):291-7.

•  Enzyme activity determination on macromolecular substrates by isothermal titration calorimetry: application to mesophilic and psychrophilic chitinases.
Lonhienne T, Baise E, Feller G, Bouriotis V, Gerday C.
Biochim Biophys Acta . 2001 Feb 9;1545(1-2):349-56.

•  Cloning, sequences, and characterization of two chitinase genes from the Antarctic Arthrobacter sp. strain TAD20: isolation and partial characterization of the enzymes.
Lonhienne T, Mavromatis K, Vorgias CE, Buchon L, Gerday C, Bouriotis V.
J Bacteriol . 2001 Mar;183(5):1773-9.

•  Chitin deacetylases: new, versatile tools in biotechnology.
Tsigos I, Martinou A, Kafetzopoulos D, Bouriotis V.
Trends Biotechnol . 2000 Jul;18(7):305-12. Review.

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