Research

The general topics of interest are:

  • Identification of novel enzymes and development of enzymatic cascades.
  • Optimization of biocatalysts via protein engineering.
  • Utilization of tailor-made nanomaterials and exploitation of their intrinsic properties during the enzymatic process.
  • Study of the enzyme function-structure relationship.
  • Process development.

Our running projects are focused on:

  • substrate scope and promiscuity of natural product (S)-adenosyl-L-methionine (SAM) dependent methyl transferases.
  • establishment of an efficient regeneration cascade of SAM cofactor.
  • enzymatic production of furane-based polymers.
  • exploring the substrate scope and enantioselectivity mechanism of cutinases.
  • valorization of olive mill wastewaters by microbial and enzymatic treatment for the production and isolation of bioactive compounds.

The Enzyme Technology laboratory was founded in February 2018 at the Department of Chemistry, University of Crete, Greece. It supports both educational and research activities of the Department and is in direct connection with the two other laboratories of Biochemistry Devision. Our laboratories have the infrastructure for Biosafety Level 1 (BSL1) experiments, fully equipped for the recombinant expression of enzymes and enzyme characterization (laminar flow bench, autoclave, thermocyclers, chambers for agarose gel electrophoresis and polyacrylamide gel electrophoresis, shakers, incubators, thermoshakers, french press, sonicator, protein purification system etc). Concerning analytics, we have 2 microtiter plate readers, 1 spectrophotometer, while we have access to gas chromatography with FID detector, HPLC with UV detector, and the MS and NMR facilities of the Department.

Group members

Lyn Lisette Kailing

PhD student (jointed with Uni Kassel)
kailing@uni-kassel.de
+49 (0)561 804 4232

Ioannis Metaxas

PhD student
giannis.metaxas93@gmail.com
+30 2810 54-5143

Jonas Peterle

PhD student (jointed with Uni Kassel)
jpeterle@uni-kassel.de
+49 (0)561 804 4232

Qing-Yun Tang

PhD student (jointed with Uni Greifswald)
qingyun.tang@stud.uni-greifswald.de
+49 (0)3834 420 4419

Panagiotis Kelefiotis

MSc student
chemp991@edu.chemistry.uoc.gr
+30 2810 54-5143

Thalia Sakoleva

MSc student
chemp1001@edu.chemistry.uoc.gr
+30 2810 54-5143

Athanasia Dimitrakouli

BSc student
chem2209@edu.chemistry.uoc.gr
+30 2810 54-5143

Michael Petrou

BSc student
chem2307@edu.chemistry.uoc.gr
+30 2810 54-5143

Eirini Prinea

BSc student
chem1967@edu.chemistry.uoc.gr
+30 2810 54-5143

Recent Publications

More Publications

Author metrics (source Scopus, 19 September 2019)

  • 35 publications in peer-review international journals
  • 7 chapters in international books (after peer-review)
  • 914 Citations
  • h-factor: 16

. Specific Residues Expand the Substrate Scope and Enhance the Regioselectivity of a Plant O???Methyltransferase. ChemCatChem, 11(14):3227-3233 | Young Researchers Series, 2019.

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. The challenge of using isopropylamine as amine donor in transaminase catalysed reactions. Organic and Biomolecular Chemistry, 17: 1634-1642, 2019.

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. Nanostructured modified ultrananocrystalline diamond surfaces as immobilization support for lipases. Diamond & Related Materials, 90: 32-39, 2018.

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. Cloning, expression and characterization of cold active esterase (EstN7) from Bacillus cohnii strain N1: A novel member of family IV. International Journal of Biological Macromolecules, 120: 1247-1255, 2018.

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. Revealing cutinases' capabilities as enantioselective catalysts. ACS Catalysis, 8: 7944-7951, 2018.

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. Isopropylamine as amine donor in transaminase-catalyzed reactions: Better acceptance through reaction and enzyme engineering. ChemCatChem, 10(18): 3943-3949, 2018.

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. S-Adenosyl-L-Homocysteine hydrolase inhibition by a synthetic nicotinamide cofactor biomimetic. Frontiers in Microbiology, 9: 505, 2018.

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. Chapter 5: Computational techniques for efficient biocatalysis. in Modern Biocatalysis: Advances Towards Synthetic Biological Systems (Eds. G. Williams & M. Hall), RSC Catalysis Series, pp. 119-152, ISBN: 978-1-78262-726-5, 2018.

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. Chapter 48: Identification and evolution of biocatalysts of interest. in Advanced Nanotechnologies for Detection and Defence Against CBRN Agents (Eds. P. Petkov et al.), NATO Science for Peace and Security Series B: Physics and Biophysics, Springer Netherlands, pp. TBA, ISBN: 978-94-024-1297-0, 2018.

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. A coupled photometric assay for characterization of S-adenosyl-L-homocysteine hydrolases in the physiological hydrolytic direction. New Biotechnology, 39: 11-17, 2017.

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Teaching

XHM060: Enzyme Technology

Elective course - Winter Semester BSc Chemistry - 6 ECTS
Lecture: Monday 9-11, Hall: A3
Seminar: Tuesday 13-15, Hall: A3

Contents of lectures:

  • Introduction to enzymology (definition, onomatology, building blocks, motifs, activity, stability etc)
  • Catalytic mechanisms and co-factors
  • Factors that affect enzyme stability
  • Enzyme kinetics and regulation of enzyme activity (inhibition/activation)
  • Biocatalyst types and formulation
  • Enzyme production and downstream processing
  • Biocatalyst immobilization
  • Bioprocesses and bioreactor types
  • Biocatalysis in non-conventional media
  • Biocatalytic strategies
  • Enzymatic cascades
  • Biocatalytic applications - white biotechnology

Contents of seminar:

  • Methods of cloning
  • Genome editing methods
  • Expression systems
  • Methods of cell lysis and sterilization
  • Methods of protein purification
  • Protein Engineering Principles
  • Analytics for determination of enzymatic activity
  • Methods of structure determination

??????89: Protein Engineering

Elective class - Spring Semester MSc Chemistry - 10 ECTS
Lecture: TBA, Hall: TBA
Seminar: TBA, Hall: TBA

Contents:

  • Principles of enzymatic catalysis and analytical chemistry of enzymatic reactions
  • Sources of novel enzymes and innovative enzymatic reactions
  • Goals of protein engineering (activity, selectivity, stability, compatibility etc)
  • Directed evolution vs. rational design
  • Catalytic promiscuity and mechanisms
  • Error-prone PCR (principles and parameters)
  • High-throughput analytics
  • Computational protein engineering
  • Methods for rapid directed evolution
  • Role of B-factor in enzyme stabilization
  • Cyclic permutation of proteins
  • Chimeric proteins production
  • Protein engineering with phage display
  • Change of substrate scope and stereoselectivity of lipases and esterases
  • Change of selectivity towards the used co-factor

Seminar:

  • Presentation of selected articles on protein engineering.

Contact