LABORATORY OF ENZYME TECHNOLOGY

Professor Yannis D. Clonis

Degrees:

Doctor of Philosophy (Ph.D.), University of Southampton, England, 1981.

Master of Science (M.Sc.), University of Warwick, England, 1976.

Bachelor (Biology), University of Athens, Greece, 1975.

Affiliation:

Director of the Laboratory of Enzyme Technology, Agricultural University of Athens.

Contact details:

Phone: +30. 210.5294311, fax: +30.210.5294307, e-mail: clonis@aua.gr

Address:

Laboratory of Enzyme Technology

Department of Agricultural Biotechnology,

Agricultural University of Athens, 

75 Iera Odos Street, GR-118 55 Athens, Greece.

Research Interests: 

Downstream processing. Development of purification processes for rapid production of high-value enzymes. Enzyme purification protocols based on novel affinity materials.

Affinity ligand design. Design, synthesis and study of biomimetic affinity ligands employing molecular modelling and biocomputing. Synthesis and application of novel affinity adsorbents. Computer-aided drug design for agricultural applications.

Immobilised enzymes and biosensors. Development of bioatalytic materials suitable for bioassays and construction of biosensors. Application to the determination of metabolites and agrochemicals.

Protein engineering and modification. Study and modification of the protein/enzyme structure. Creation of biocatalysts with new properties employing recombinant DNA technology. 

Other Activities:

Member of the editorial board of the scientific journals:

Applied Biochemistry and Biotechnolgy (Humana, USA),

Bioseparation (Kluvwer, NL),

Process Biochemistry (Elsevier, NL).

- Delegate of the Downstream Processing Task Group of the European Federation of

Biotechnology.

- Member of the Planning Committee of the International Society of Molecular Recognition.

- Member of the Hellenic Society of Biochemistry & Molecular Biology.

- Member of the Hellenic Society of Biotechnology.

- Member of the Hellenic Biological Society.

Publications:

Research papers

R1. Affinity Chromatography of Immobilized Nucleotides. The Synthesis, Specificity and Applications of Immobilized Inosine 5'-monophosphate (IMP). 

Y.D. CLONIS and C.R. LOWE.

Eur. J. Biochem., 110 (1980) 279-288. 

R2. Triazine Dyes: A New Class of Affinity Labels for Nucleotide-dependent Enzymes. 

Y.D. CLONIS and C.R. LOWE.

Biochem. J., 191 (1980) 247-251. 

R3. Affinity Chromatography on Immobilized Triazine Dyes. Studies on the Interaction with Multinucleotide-dependent Enzymes. 

Y.D. CLONIS and C.R. LOWE.

Biochim. Biophys. Acta, 659 (1981) 86-98. 

R4. The Interaction of Yeast Hexokinase with Procion Green H-4G. 

Y.D. CLONIS, M. GOLDFINCH and C.R. LOWE.

Biochem. J., 197 (1981) 203-211. 

R5. Affinity Chromatography on Immobilized Triazine Dyes. Post-immobilization Chemical Modification of Triazine Dyes. 

Y.D. CLONIS.

J. Chromatogr., 236 (1982) 69-80. 

R6. Affinity Chromatography on Immobilized Nucleotides. Studies on the Interaction of E. coli IMP Dehydrogenase with Immobilized Nucleotides. 

Y.D. CLONIS and M.J. GOLDFINCH.

J. Mol. Catalysis, 16 (1982) 1-9. 

R7. Process-scale High Performance Liquid Affinity Chromatography. 

Y.D. CLONIS, K. JONES and C.R. LOWE.

J. Chromatogr., 363 (1986) 31-36. 

R8. Novel Cationic Triazine Dyes in Protein Purification. 

Y.D. CLONIS, C.V. STEAD and C.R. LOWE.

Biotech. Bioengin., 30 (1987) 621-627. 

R9. High Performance Liquid Chromatography for the Purification of Restriction Endonucleases. Application to Ban II, Sac I and Sph I. 

V. BOURIOTIS, A. ZAFEIROPOULOS and Y.D. CLONIS.

Analyt. Biochem., 160 (1987) 127-134. 

R10. Dye-ligand Chromatography for the Resolution and Purification of Restriction Endonucleases. 

G. VLATAKIS, G. SKARPELIS, I. STRATIDAKI, V. BOURIOTIS and Y.D. CLONIS.

Applied Biochem. and Biotech., 15 (1987) 201-212. 

R11. Matrix Evaluation for Preparative High Performance Affinity Chromatography. 

Y.D. CLONIS

J. Chromatogr., 407 (1987) 179-188. 

R12. Preparative Affinity Precipitation of L-Lactate Dehydrogenase. 

J. PEARSON, Y.D. CLONIS and C.R. LOWE.

J. Biotechnol., 11 (1989) 267-274. 

R13. Purification of the Glycoprotein Allergen Ag7 from Mugwort Pollen by Concanavalin A Affinity Chromatography. 

B. NILSEN, B. PAULSEN, Y.D. CLONIS, K.S. MELLBYE.

J. Biotechnol., 16 (1990) 305-316. 

R14. Monosized Adsorbents for High performance Affinity Chromatography. Application to the Purification of Calf Intestinal Alkaline Phosphatase and Human Urine Urokinase. 

Y.D. CLONIS and C.R. LOWE.

J. Chromatogr., 540 (1991) 103-111. 

R15. Downstream Processing of Diagnostic Enzymes. Optimised Protocols for the Simultaneous Separation and Purification of Lactate Dehydrogenase and Pyruvate Kinase from Rabbit Muscle. 

G. TSAMADIS, N. PAPAGEORGAKOPOULOU and Y.D. CLONIS.

Bioprocess Engin., 7 (1992) 213-218. 

R16. Simultaneous Separation and Purification of Pyruvate Kinase and Lactate Dehydrogenase by Dye-ligand Chromatography. 

T. MAKRIYANNIS and Y.D. CLONIS.

Process Biochem., 28 (1993) 179-185. 

R17. The Interaction of Candida boidinii Formate Dehydrogenase with a New Family of Chimeric Biomimetic Dye-Ligands. 

N. LABROU and Y.D. CLONIS

Arch. Biochem. Biophys. 316 (1995) 169-178. 

R18. Biomimetic Dye-Ligands for Oxalate-Recognizing Enzymes. Studies with Oxalate Oxidase and Oxalate Decarboxylase. 

N. LABROU and Y.D. CLONIS

J. Biotechnol., 40 (1995) 59-70. 

R19. Oxaloacetate Decarboxylase: on the Mode of Interaction with Substrate-Mimetic Affinity Ligands. 

N. LABROU and Y.D. CLONIS

Arch. Biochem. Biophys. 321 (1995) 61-70. 

R20. Biomimetic-dye Affinity Adsorbents for Enzyme Purification. Application to the One-step Purification of Candida boidinii Formate Dehydrogenae. 

N.E. LABROU, A. KARAGOUNI and Y.D. CLONIS

Biotech. Bioengin., 48 (1995) 278-288. 

R21. Biomimetic-dye affinity chromatography for the Purification of Bovine Heart Lactate Dehydrogenase 

N.E. LABROU and Y.D. CLONIS

J. Chromatogr., 718 (1995) 35-44. 

R22. Biomimetic-dye Affinity Chromatography for the Purification of Mitochondrial L-Malate Dehydrogenase from Bovine Heart. 

N.E. LABROU and Y.D. CLONIS

J. Biotechnol., 45 (1996) 185-194. 

R23. Dye Affinity Labelling of Bovine Heart Mitochondrial Malate Dehydrogenase and Study of the NADH-binding Site. 

N.E. LABROU, E. ELIOPOULOS and Y.D. CLONIS

Biochem. J., 315 (1996) 687-693. 

R24. Molecular Modelling for the Design of Chimeric Biomimetic Dye-ligands and their Interaction with Bovine Heart Mitochondrial Malate Dehydrogenase. 

N.E. LABROU, E. ELIOPOULOS and Y.D. CLONIS

Biochem. J., 315 (1996) 695-703. 

R25. Simultaneous Purification of L-Malate Dehydrogenase and L-Lactate Dehydrogenase from Bovine Heart by Biomimetic-dye Affinity Chromatography. 

N.E. LABROU and Y.D. CLONIS.

Bioprocess Engin., 16 (1997) 157-161. 

R26. Design and Study of Peptide-ligand Affinity Adsorbents. Application to the Case of Trypsin Purification from Bovine Pancreas. 

T. MAKRIYANNIS and Y.D. CLONIS

Biotech. Bioengin., 53 (1997) 49-57. 

R27. L-Malate Dehydrogenase from Pseudomonas studzeri: Purification and Characterization. 

N.E. LABROU and Y.D. CLONIS

Arch. Biochem. Biophys. 337 (1997) 103-114. 

R28. Oxalate Oxidase from Barley Roots: Purification to Homogeneity and Study of some Molecular, Catalytic and Binding Properties. 

V. KOTSIRA and Y.D. CLONIS

Arch. Biochem. Biophys. 340 (1997) 239-249. 

R29. Purification of Alcohol Dehydrogenase from Four Genotypes of the Olive Fruit Fly Bactrocera (Dacus) oleae.

V.E. MAZI, N. COSMIDIS, Y.D. CLONIS and M. LOUKAS

Biotechnol. Progr., 14 (1998) 294-299. 

R30. Biochemical Differences Between Products of the ADH Locus in Olive Fruit Fly (Bactrocera oleae).

V.E. MAZI, N. COSMIDIS, Μ. LOUKAS, Y.D. CLONIS and E. ZOUROS. 

Biochem. Genet. 36 (1998) 259-269.

R31. Chemical Μodification of Βarley Root Οxalate Οxidase Shows the Presence of a Lysine, a Carboxylate, and Disulfides, Essential for Enzyme Activity.

V. KOTSIRA and Y.D. CLONIS

Arch. Biochem. Biophys. 356 (1998) 117-126. 

R32. Colorimetric Assay for Lecithin Using Two Co-immobilized Enzymes and an Indicator Dye Conjugate. 

V. KOTSIRA and Y.D. CLONIS

J. Agric. Food Chem. 46 (1998) 3389-3394. 

R33. Molecular Modeling for the Design of a Biomimetic Chimeric Ligand. Application to the Purification of Bovine Heart L-Lactate Dedydrogenase.

N.E. LABROU, E. ELIOPOULOS and Y.D. CLONIS

Biotech. Bioengin., 63 (1999) 322-332. 

R34. Oxaloacetate Decarboxylase from Pseudomonas studzeri: Purification and Characterization.

N.E. LABROU and Y.D. CLONIS

Arch. Biochem. Biophys., 365 (1999) 17-24.

R35. Growth of Candida boidinii in a Methanol-Limited Continuous Culture and the formation of Methanol-Degrading Enzymes.

G. AGGELIS, S. FAKAS, S. MELISSIS and Y.D. CLONIS

J. Biotechnol., 72 (1999) 127-139.

R36. Characterization of the NAD Binding Site of Candida boidinii Formate Dehydrogenase by Affinity Labeling and Site Directed Mutagenesis.

N.E. LABROU, D.J. RIGDEN and Y.D. CLONIS

Eur. J. Biochem., 267 (2000) 6657-6664.

R37. Structural and functional role of the glutathione binding residues in maize glutathione S-transferase.
N.E. LABROU, L.V. MELLO and Y.D. CLONIS
Biochem. J., (2001) 358, 101-10.

R38. The conserved Asn49 in maize glutathione S-transferase modulates substrate binding, catalysis and intrasubunit communication.
N.E. LABROU, L.V. MELLO and Y.D. CLONIS
Eur. J. Biochem., (2001) 268, 3950-7.

R39. A New Family of Glutathionyl-Biomimetic Ligands for Affinity Chromatography of Glutathione-Recognozing Enzymes

S. MELISSIS, D.J. RIGDEN and Y.D. CLONIS

J. Chromatogr., (2001) 917, 29-43.

E40. A portable fiber-optic pesticide bionsensor based on immobilized cholinesterase and sol-gel entrapped bromocresol purple for in-field use.
V.G. ANDREOU and Y.D. CLONIS
Biosensors & Bioelectronics, 17 (2001) 61-69.

R41. Novel fiber-optic biosensor based on immobilized glutathione S-transferase and sol-gel entrapped bromocresol green for the determination of atrazine.
V.G. ANDREOU and Y.D. CLONIS
Analyt. Chim. Acta, 460 (2002) 151-161.

R42. Galactosyl-biomimetic dye-ligands for the purification of Dactylium dendroides galactose oxidase.

C.F. MAZITSOS, D.J. RIGDEN, P.G. TSOUNGAS and Y.D. CLONIS.

J. Chromatogr. A, 954 (2002) 137-150.

R43. Galactosyl-mimodye ligands for Pseudomonas fluorescens β-galactose dehydrogenase: design, synthesis and evaluation.
C.F MAZITSOS, D.J. RIGDEN, P.G. TSOUNGAS and Y.D. CLONIS
Eur. J. Biochem. 269 (2002) 5391-5405.

R44. Designed chimaeric galactosyl-mimodye ligands for the purification of Pseudomonas fluorescens β-galactose dehydrogenase.
C.F MAZITSOS, D.J. RIGDEN and Y.D. CLONIS
J. Chromatog. A, 1029 (2004) 103-112.

R45. Engineering the pH-dependence of kinetic parameters of maize glutathione S-transferase I by site-directed mutagenesis.
N.E. LABROU, D.J. RIGDEN and Y.D. CLONIS
Biomolecular Engineering, 21 (2004) 61-66.

R46. Interaction of L-glutamate oxidase with triazine dyes: selection of ligands for affinity chromatography.
N.E. KATSOS, N.E. LABROU and Y.D. CLONIS
J. Chromatogr. B, 807 (2004) 277-285.

Reviews & chapters in books

1. Some Preparative and Analytical Applications of Triazine Dyes. 

C.R. LOWE, Y.D. CLONIS, M.J. GOLDFINCH, D.A.P. SMALL and T. ATKINSON.
Analyt. Chem. Symp. Series, 9 (1982) 389-398.

2. General-ligand Affinity Chromatography. 

Y.D. CLONIS.
Chimika Chronika, 11 (1982) 87-110.

3. Affinity Chromatography. 

C.R. LOWE and Y.D. CLONIS.
Bioactive Polymeric Systems, 1985, pp. 203-222, Plenum, New York, USA.

4. Bioactive Polymers: Immobilized Enzymes and Cells. 

Y.D. CLONIS.
Chimika Chronica, 51 (1986) 374-378.

5. The Design and Applications of Biomimetic Dyes in Biotechnology. 

C.R. LOWE, S. BURTON, J. PEARSON, Y.D. CLONIS and C.V. STEAD.
J. Chromatogr., 376 (1986) 121-130.

6. Dye-ligand Chromatography. 

Y.D. CLONIS.
Reactive Dyes in Protein and Enzyme Technology, 1987, pp. 33-49, Macmillan, Basingstoke, U.K.

7. High Performance Dye-ligand Chromatography. 

Y.D. CLONIS and D.A.P. SMALL.
Reactive Dyes in Protein and Enzyme Technology, 1987, pp. 87-100, Macmillan, Basingstoke, U.K.

8. Large-scale Affinity Chromatography. 

Y.D. CLONIS.
Bio/Technology, 5 (1987) 1290-1293.

9. The Applications of Reactive Dyes in Protein and Enzyme Downstream Processing. 

Y.D. CLONIS.
CRC Crit. Rev. Biotechnol., 7 (1988) 263-279.

10. Affinity Chromatography. 

Y.D. CLONIS and C.R. LOWE.
Principles of Clinical Biochemistry, vol. 2, 1988, pp. 383-390, Heinemann, Oxford, U.K.

11. High Performance Affinity Chromatography. 

Y.D. CLONIS.
HPLC-A Practical Approach, 1989, pp. 157-182, IRL Press, Oxford, U.K.

12. Biomimetic Dyes in Biotechnology. 

C.R. LOWE, N. BURTON, S. DILMAGHANIAN, S. McLOUGHLIN, J. PEARSON, D. STEWART and Y.D. CLONIS.
Protein-Dye Interactions: Developments and Applications, 1989, pp. 11-20, Elsevier Science Publishers, Barking, Essex, U.K.

13. Biotechnology in Greece. 

Y.D. CLONIS.
Int. Industrial Biotechnol., 9 (1989) 20-21.

14. Process Affinity Chromatography. 

Y.D. CLONIS.
Separation Processes in Biotechnology, 1990, pp. 401-445, Marcel-Dekker, New York, USA.

15. Preparative Dye-ligand Chromatography. 

Y.D. CLONIS.
HPLC of Proteins, Peptides and Polynucleotides, 1991, pp. 453-468, VCH Publishers, New York, USA.

16. High Performance Affinity Chromatography for Protein Separation and Purification. 

Y.D. CLONIS.
Practical Protein Chromatography, Humana Press Inc., New Jersey, USA, 11 (1992) 105-123.

17.Affinity Methods in Bioseparation 

Y.D. CLONIS
Proceedings of the 6th European Congress on Biotechnology, Florence, Italy, June 13-17, 1993, pp. 527-533.

18. The Affinity Technology in Downstream Processing. 

N. LABROU and Y.D. CLONIS
J. Biotechnol., 36 (1994) 95-119. 

19. Peptide Affinity Ligands in Protein Purification Technology.

Y.D. CLONIS
Hellenic Forum on Bioactive Peptides, Crete University Press, 1997, pp. 121-128.

20. The Tachykinin Family of Peptides and Their Receptors.

N.E. LABROU and Y.D. CLONIS
Bioactive Peptides in Drug Discovery and Design: Medical Aspects (I. Matsoukas & T. Mavromoutsakos, eds.), IOS Press, Amsterdam, The Netherlands, 1999, pp. 225-233.

21. Affinity separation: dye-ligands.

Y.D. CLONIS

Encyclopaedia of Separation Science (I. Wilson, ed.), Academic Press Ltd., London, UK, 2000, pp. 259-265.

22.Biomimetic dyes as affinity chromatography tools in enzyme purification.

Y.D. CLONIS, N.E. LABROU, V.Ph. KOTSIRA, C. MAZITSOS, S. MELISSIS & G. GOGOLAS

J. Chromatogr. A, 891 (2000) 33-44.

23. Immobilised Synthetic Dyes in Affinity Chromatography.

N.E. LABROU and Y.D. CLONIS

Biochromatography - Theory and Practice (M.A. Vijayalakshmi, ed.), Taylor and Francis Publishers, London, Chap. 8, pp 235-251, 2002.

Book and Volume publications

1. Reactive Dyes in Protein and Enzyme Technology.

Y.D. CLONIS, A. ATKINSON, C.J. BRUTON and C.R. LOWE (editors).

Macmillan Press Ltd., Basingstoke, U.K., 1987.

2. Downstream Processing.

Y.D. CLONIS (editor).

J. Biotechnol., 11 (2,3), (1989) 89-289.

3. Enzyme Biotechnology (Book in Greek)

Y.D. CLONIS (author).

Crete University Press, Heraklion, Crete, Greece, 1997. 

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