LABORATORY OF ENZYME TECHNOLOGY


ΕΡΓΑΣΤΗΡΙΟ ΕNΖΥΜΙΚHΣ ΤΕΧΝΟΛΟΓΙΑΣ

 

PUBLICATIONS - ΔΗΜΟΣΙΕΥΣΕΙΣ

 

Α. Downstream Processing, Design and Application of Biomimetic Ligands.

 

Α. Κατιούσα Επεξεργασία, Σχεδιασμός και Εφαρμογή Βιομιμητικών Μορίων.

 

 

1. Affinity Methods in Bioseparation.

Y.D. CLONIS

Proceedings of the 6th European Congress on Biotechnology, Florence, Italy, June v.13-17 (1993) pp. 527-533.

 

2. 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.

 

3. The Affinity Technology in Downstream Processing.

N.E. LABROU and Y.D. CLONIS

J. Biotechnol., 36 (1994) 95-119.

 

4. Τhe Interaction of Candida boidinii Formate Dehydrogenase with a New Family of chimeric Biomimetic Dye-Ligands.

N.E. LABROU and Y.D. CLONIS

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

 

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

N.E. LABROU and Y.D. CLONIS

J. Biotechnol., 40 (1995) 59-70.

 

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

N.E. LABROU and Y.D. CLONIS

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

 

7. Biomimetic-dye Affinity Chromatography for the Purification of Bovine Heart Lactate Dehydrogenase

N.E. LABROU and Y.D. CLONIS

J. Chromatogr. A, 718 (1995) 35-44.

 

8. 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

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

 

9. 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.

 

10. 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.

 

11. Peptide Affinity Ligands in Protein Purification Technology.

Y.D. CLONIS

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

 

12. 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.

 

13. 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.

 

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

N.E. LABROU and Y.D. CLONIS

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

 

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

T. MAKRIYANNIS and Y.D. CLONIS

Biotechnol. Bioengin., 53 (1997) 49-57.

 

16. 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.

 

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

N.E. LABROU and Y.D. CLONIS

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

 

18. 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

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

 

19. Affinity Separation: Dye-Ligands.

Y.D. CLONIS

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

 

20. Biomimetic Dyes as Affinity Chromatography Tools in Enzyme Purification.

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

J. Chromatogr. A, 891 (2000) 33-44.

 

21. Improved Purification of Candida boidinii Formate Dehydrogenase.

N.E. LABROU

Bioseparation, 9 (2000) 99-104

 

22. Dye-ligand affinity chromatography for protein separation and purification.

N.E. LABROU

Methods in Molecular Biology:Affinity Chromatography: Methods and Protocols’. (J.M. Walker, ed.), Humana Press, USA, 2000, v.147, pp. 129-139.

 

23.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. A, 917 (2001) 29-43.

 

24. Dye-ligand affinity adsorbents for enzyme purification.

N.E. LABROU

Appl. Biochem. Biotechnol., Part B : Mol. Biotechnol., 20 (2002) 77-84

 

25. 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, 2002, Chap. 8, pp. 235-251.

 

26. 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.

 

27. Affinity chromatography.

N.E. LABROU

Methods for Affinity-Based Separations Of Enzymes and Proteins, (M.N., Gupta, ed.), Birkhϊuser Verlag AG, Switzerland, 2002, 16-28.

 

28.  Dye-ligand affinity adsorbents for enzyme purification.
N.E. LABROU

Mol. Biotechnol., 20 (2002) 77-84.

 

29. Dye-ligand and biomimetic affinity chromatography.

N.E. Labrou, K. Mazitsos and Y. Clonis

In: D.S. Hage (Ed.), Handbook of Affinity Chromatography. Marcel Dekker, Inc, New York, (in press).

 

30. 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.

 

31. Design and selection of ligands for affinity chromatography.
N.E. LABROU

J. Chromatogr. B, 790 (2003) 67-78.

32. 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.

33. 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.

34. Affinity chromatography matures as bioinformatic and combinatorial tools develop. 

Y.D. CLONIS

J. Chromatogr. A, 1101 (2006) 1-24.

 

35. Lock-and-key motif as a concept for designing affinity adsorbents for protein purification. 

D. PLATIS, C.A. SOTRIFFER, Y.D. CLONIS, AND N.E. LABROU

J. Chromatogr. A, 1128 (2006) 138-151

 

36. Nucleotide-mimetic synthetic ligands for DNA-recognizing enzymes. One-step purification of Pfu DNA polymerase. 
S.
Melissis, N.E. Labrou, Y.D. Clonis

J. Chromatogr. A , 1122 (2006) pp. 63-75

 

37. Development of an aqueous two-phase partitioning system for fractionating therapeutic proteins from tobacco extract 

D. PLATIS, N.E. LABROU

J.  Chromatogr. A, 1128 (2006) 114-124.

 

38. Development of recombinant protein-based influenza vaccine. Expression and affinity purification of H1N1 influenza virus neuraminidase. 

T. DALAKOURAS, B.J. SMITH, D. PLATIS, M.M.J. COX, N.E. LABROU

J. Chromatogr. A, 1136 (2006) 48-56

 

39. One-step purification of Taq DNA polymerase using nucleotide-mimetic affinity chromatography.

S. Melissis, N.E. Labrou, Y.D. Clonis

Biotechnol. J., 2 (2007) 121-132.

 

40. Affinity chromatography for the purification of therapeutic proteins from transgenic maize using immobilized histamine 

D. PLATIS, N.E. LABROU

J. Separation Sci., 31 (2008) 636-645.

 

41. Cost-effective production of a vaginal protein microbicide to prevent HIV transmission.

K. Ramessar, T. Rademacher, M. Sack, J. Stadlmann, D. Platis, G. Stiegler, N.E. Labrou, F. Altmann, J. Ma, E. Stöger, T. Capell, P. Christou

Proc Natl Acad Sci U S A. 105 (2008) 3727-32

 

42. New downstream processing strategy for the purification of monoclonal antibodies from transgenic tobacco plants.

D. Platis, J. Drossard, R. Fischer, J.K. Ma, N.E. Labrou

J Chromatogr A, 1211 (2008) 80-89.

 

43. Application of a PEG/salt aqueous two-phase partition system for the recovery of monoclonal antibodies from unclarified transgenic tobacco extract.

D. Platis, N.E. Labrou

Biotechnol J., 4 (2009) 1320-1327.

 

44. Combinatorial de novo design and application of a biomimetic affinity ligand for the purification of human anti-HIV mAb 4E10 from transgenic tobacco.

D. Platis, A. Maltezos, J.K. Ma, N.E. Labrou

J. Mol. Recognit., in press

 

45. Purification of Moloney Murine Leukemia virus reverse transcriptase lacking RNAse activity (M-MLVH -RT) on a 9-aminoethyladenine-[1,6-diamine-hexane]-triazine, selected from a combinatorial library of dNTP-mimetic ligands.

S. Melissis, A. Papageorgiou, N.E. Labrou, Y.D. Clonis

J. Chromatogr. Sci., in press.

 

---

 

Β. Enzyme Engineering.

 

Β. Πρωτεϊνική Μηχανική

 

 

1. Dye Affinity Labeling 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.

 

2. Chemical Modification of Barley Root Oxalate Oxidase 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.

 

3. 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.

 

4. 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.

 

5. Afinity Labeling of Oxaloacetate Decarboxylase by Novel Dichlorotriazine Linked α-Ketoacids.

N.E. LABROU

Protein J., 18 (1999) 729-733.

 

6. Dye Affinity Labelling of Yeast Alcohol Dehydrogenase.

N.E. LABROU

J. Enz. Inhibition, 15 (2000) 487-496.

 

7. 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.

 

8. Active Site Characterization of Candida boidinii Formate Dehydrogenase.

N.E. LABROU and D.J. RIGDEN

Biochem. J., 354 (2001) 455-463.

 

9. Functional and structural and role of the glutathione binding residues in maize glutathione S-transferase.
N.E. LABROU, L.V. MELLO and Y.D. CLONIS
Biochem. J., 358 (2001) 101-1
10.

 

10. 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., 268 (2001) 3950-3957.

 

11.Glutathione S-transferase: de novo design of glutathionyl-analogues and combinatorial design of novel enzyme reactions.

N.E. Labrou, Axarli, I., Melissis, S., Kotzia, G., Katsos, N. & Y.D. Clonis.

3rd Forum on Bioactive Peptides. Ed.: P. Kordopatis, Typorama, Patra, 2003, 249-258.

 

12. The structure-function relationship in the clostripain family of peptidases. 

N.E. Labrou, D.J. Rigden

Eur.J. Biochem., 271 (2004) 983-992.

 

13. 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
Biomol. Engine., 21 (2004) 61-66.

 

14. S-(2,3-dichlorotriazinyl)glutathione: A new affinity label for probing the structure and function of glutathione transferases
G.A.
Kotzia, N.E. Labrou

Eur. J. Biochem., 271 (2004) 3503-3511

 

15. Characterization of the ligandin site of maize glutathione S-transferase I. 

I.A. Axarli, D.J. Rigden, N.E. Labrou

Biochem. J., 382 (2004) 885-893

 

16. Engineering the xenobiotic substrate specificity of maize glutathione S-transferase I.
N.E.
Labrou, G.A. Kotzia, G.A., Y.D. Clonis

Protein Engine. Des. Select., 17 (2004) 741-748

 

17. Directed enzyme evolution: bridging the gap between natural enzymes and commercial applications.  
N.E. LABROU

Biomol. Engine., 22 (2005), pp. vii-ix

 

18. Engineering the substrate specificity of cytochrome P450 CYP102A2 by directed evolution: production of an efficient enzyme for biotransformation of fine chemicals.

I. AXARLI, A. PRIGIPAKI, N.E. LABROU

Biomol. Engin., 22 (2005) 81-88.

 

19. Kinetic analysis of maize glutathione S-transferase I catalysing the detoxification from chloroacetanilide herbicides

N.E. Labrou, M. Karavangeli, A. Tsaftaris, A., Y.D. Clonis,

Planta, 222 (2005) 91-97.

 

20. Development of transgenic tobacco plants overexpressing maize glutathione S-transferase I for chloroacetanilide herbicides phytoremediation

M. Karavangeli, N.E. Labrou, Y.D. Clonis, A. Tsaftaris

Biomol. Engine., 22 (2005) 121-128.

 

21. Cloning, expression and characterisation of Erwinia carotovora L-asparaginase.

G.A. Kotzia, N.E. Labrou

J. Biotechnol., 119 (2005) 309-323.

 

22. Structure of a calcium-deficient form of influenza virus neuraminidase: Implications for substrate binding.

B.J. Smith, T. Huyton, R.P. Joosten, J.L. McKimm-Breschkin, J.-G. Zhang, C.S. Luo, M.-Z. Lou, (...), T.P.J. Garrett

Acta Crystallogr. (Section D: Biol. Crystallogr.), 62 (2006) 947-952.

 

23. Structure-guided design of a novel class of benzyl-sulfonate inhibitors for influenza virus neuraminidase.

D. Platis, B.J. Smith, T. Huyton, N.E. Labrou

Biochem. J., 399 (2006) 215-223.

 

24. L-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization.

G.A. Kotzia, N.E. Labrou

J. Biotechnol., 127 (2007) 657-669.

 

25. Tailoring structure-function properties of L-asparaginase: engineering resistance to trypsin cleavage.

G.A. Kotzia, K. Lappa, N.E. Labrou

Biochem. J., 404 (2007) 337-343.

 

26. A dehydrochlorinase-based pH change assay for determination of DDT is sprayed surfaces.

E. Morou, H.M. Ismail, A.J. Dowd, J. Hemingway, N.E. Labrou, M. Paine, J. Vontas

Anal Biochem., 378 (2008) 60-64

 

27. Structure-guided alteration of coenzyme specificity of formate dehydrogenase by saturation mutagenesis to enable efficient utilization of NADP+ 
A.
Andreadeli, D. Platis, V. Tishkov, V. Popov, N.E. Labrou

FEBS Journal, 275 (2008) 3859-3869.

28. Chemical and genetic engineering strategies to improve the potency of pharmaceutical proteins and enzymes.

D. Platis, D., N.E. Labrou

Curr. Med. Chem., 15 (2008) 1940-1955.

 

29. Crystallographic and Functional Characterization of the Fluorodifen-inducible Glutathione Transferase from Glycine max Reveals an Active Site Topography Suited for Diphenylether Herbicides and a Novel L-site.

I. Axarli, P. Dhavala, A. Papageorgiou, N.E. Labrou

J. Mol. Biol., 385 (2009) 984-1002.

 

30. Engineering thermal stability of l-asparaginase by in vitro directed evolution.

G.A. Kotzia, N.E. Labrou

FEBS Journal, 276 (2009) 1750-1761.

 

31. Sulphonamide-based bombesin prodrug analogues for glutathione transferase, useful in targeted cancer chemotherapy.

I. Axarli, N.E. Labrou, C. Petrou, N. Rassias, P. Cordopatis, Y.D. Clonis

Eur. J. Med.  Chem., 44 (2009) 2009-2016.

 

32. Cloning and characterization of Lotus japonicus formate dehydrogenase: A possible correlation with hypoxia.

A. Andreadeli, E. Flemetakis, I. Axarli, M. Dimou, M.K. Udvardi, P. Katinakis, N.E. Labrou

Biochim. Biophys. Acta, 1794 (2009) 976-984.

 

33. Molecular and biochemical characterization of the parvulin-type PPIases in Lotus japonicus.

E.D. Kouri, N.E. Labrou, S.D. Garbis, K.I. Kalliampakou, C. Stedel, M. Dimou, M.K. Udvardi, P. KATINAKIS, E. Flemetakis

Plant Physiol., 150 (2009) 1160-1173.

 

34. Crystal structure of Glycine max glutathione transferase in complex with glutathione: investigation of the mechanism operating b the Tau class glutathione transferases.

I. Axarli, P. Dhavala, A.C. Papageorgiou, N.E. Labrou

Biochem J., 422 (2009) 247-256.

 

35. Glutathione transferases: emerging multidisciplinary tools in red and green biotechnology.

E.G. Chronopoulou, N.E. LABROU

Recent Pat Biotechnol., in press.

 

 

 

C. Immobilized Enzymes and Biosensors.

 

Γ. Ακινητοποιημένα Ένζυμα και Βιοαισθητήρες

 
 

1. 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.

2. A portable fiber-optic pesticide biosensor based on immobilized cholinesterase and sol-gel entrapped bromcresol purple for in-field use.

V.G. ANDREOU and Y.D. CLONIS
Biosens. 
Bioelectron., 17 (2001) 61-69. 

3. Φορητός ενζυμικός βιοαιθητήρας οπτικών ινών που βασίζεται σε τεχνολογία Sol-Gel για τον προσδιορισμό οργανοφωσφορικών και καρβαμιδικών φυτοφαρμάκων σε φυσικά ύδατα.
Ε. ΑΝΔΡΕΟΥ, Θ. ΝΤΟΥΡΟΥ και Ι. ΚΛΩΝΗΣ
Πρακτικά 18ου Πανελληνίου Συνεδρίου Χημείας (Ενόργανη Ανάλυση-Διαπίστευση Εργαστηρίων), 10-13 Μαρτίου 2001, Πειραιάς, Αττική. 

4. 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.

 

5. Engineering sensitive glutathione transferase for the detection of xenobiotics.

P. Kapoli, I.A. Axarli, D. Platis, M. Fragoulaki, M. Paine, J. Hemingway, J. Vontas, N.E. Labrou

Biosens. Bioelectron., 24 (2008) 498-503.

 

 

D. Books

 

Δ. Βιβλία

1. Ενζυμική Βιοτεχνολογία

Ι.Δ. ΚΛΩΝΗΣ (συγγραφέας)

Πανεπιστημιακές Εκδόσεις Κρήτης, ΙΤΕ, Ηράκλειο, Κρήτη, 1997.

 

2. Ενζυμολογία

Ι.Δ. ΚΛΩΝΗΣ (συγγραφέας)

Εκδόσεις ΕΜΒΡΥΟ, Αιγάλεω, Αττική, 2007.

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