SCIENTIFIC AND PRODUCTION CENTER “ARMBIOTECHNOLOGY” NAS RA

soc soc soc

Research Units

 

Artur Hambardzumyan

 
Head of Laboratory
PhD in Biology
e-mail: arthambardzumyan@armbiotech.am 

 

Laboratory of Protein Technologies

 

Laboratory of Protein Technologies was founded in 2010.

 

Field of Activity

  • Molecular biology:

gene engineering, obtaining of cloning amplicons, gene expression, DNA, RNA, isolation, purification and characterization of protein and other molecules (melanin)

  • Biochemistry:

determination of physicochemical and catalytic properties of enzymes, study of mechanisms of enzymatic catalysis and statistical analysis

  • Biotechnology:

development of technologies based on strain-producers, development of technologies based on enzymes and their immobilized forms

  • Bioinformatics:

search for genes based on genome analysis, design of primers, construction of genes based on sequence data and placement in international databases, investigation of ligand-macromolecular interaction by docking analysis

  • Microbiology:

detection of new bacteria, classification of bacteria based on 16S rRNA sequencing.

  • Biophysics:

reconstruction and characterization of ionic channels in artificial bilayer lipid membranes, obtaining of a three-dimensional protein structure from homologous modeling and its application in molecular docking experiments

 

RESEARCH MAIN DIRECTIONS

  • Expression cloning of protein (enzyme) genes
  • Isolation, purification and characterization of recombinant and natural proteins (enzymes) asymmetric enzymatic synthesis of amino acids
  • Microbiological synthesis of amino acids
  • Bacterial melanin synthesis
  • Obtaining of bacterial insecticidal preparations    

 

MAIN INVESTIGATION RESULTS

  • Obtaining of recombinant strain-producers of L-arginine (E. coli - up to 35 g / l and Br. Flavum - up to 31 g / l) and study of physiological and biochemical properties of these strain-producers
  • Obtaining of strain-producers of bacterial, water-soluble, biologically active melanin (particularly plant growth stimulators)
  • Obtaining of strain-producers of L- and D-aminoacylases
  • Isolation and characterization of a number of bacterial amino acid transaminases (alanine, aspartate, aromatic)
  • Clarification of a transport nature of overproduction of certain amino acid strain-producers (L-alanine, L-proline)
  • Development of technologies for production of a number of enzymes: glucose oxidase, peroxidase, glutamate dehydrogenase, etc.
  • Characterization of potential ligands for a number of target proteins (EGFR, MAPK14, TLR4, etc.) by molecular docking    

 

MAIN OBJECTIVES AND TASKS

It is envisaged to obtain BAS and their strain-producers by genetic-selection, gene engineering, biochemical engineering and immunological methods and to study them by biochemical and immunological methods.

In particular, it is envisaged to create the objects of the following classes:

  • high purity enzymes and proteins (natural and recombinant) for analytical and practical purposes
  • various immobilized enzymes and biocatalysts with improved characteristics (activity, stability, possible activity in anhydrous or low water mixtures, etc.) for using in different types of biotransformation processes
  • monoclonal and polyclonal antibodies
  • vaccines of different generations (protein constructs and constructs of various recombinant DNAs carrying antigen epitope genes)
  • recombinant strains producing amino acids, peptides, proteins (enzymes) and vaccines    

 

LIST OF GRANT PROJECTS

  1. SCS MES RA-BRFBR 18BL-040 “Use of bacterial melanin as a stimulant for growth and development of rare pharmaceutical and local high ornamental plants”, 2018-2020 (Project Manager: A. Hovsepyan, PhD).
  2. ISTC #A‑2289 “Synthesis and screening of a new generation of optically active non-proteinogenic  α-amino acids, peptides and polymers containing unsaturated groups in the side chain”, 2017-2020 (Project Manager: Academician of NAS RA, Prof. A. Saghyan).
  3. 16GE-034 “Application of of hyperthermophil carbamoylase in enzymatic hydrolysis of N-carbamoyl-D-amino acids”, 2017-2019 (Project Manager: A. Hovsepyan, PhD).
  4. ANSEF  Plant-4720 “New high-efficiency insecticidal preparations based on the melaninogenic strains of Bacillus thuringiensis”, 2017 (Project Manager: S. Avetisyan, PhD).
  5. SCS MES RA-BRFBR-2016 ՀԲ16-53 “Preparation and application of biopreparations based on the Bacillus thuringiensis for promoting plant growth and protection against pests”, 2016-2018 (Project Manager: A. Hovsepyan, PhD).
  6. 16YR-2I068 “Targeted modification of recombinant D-hydroxinase of Geobacillus stearothermophilus to improve the catalytic properties of the enzyme”, 2016-2018 (Project Manager: H. Aganyants, PhD).
  7. 16YR-2I016 “Obtaining of the strain-producer of  α-amylase on the base of the gene of Bacillus amyloliquefaciens α-amylase and recombinant enzyme characterization”, 2016-2018 (Project Manager: A. Paloyan, PhD).
  8. ANSЕF biоtеch-4241 “Microbial melanin as a means of preclinical study of potential treatment for Parkinson’s disease using neurodegeneration models”, 2016 (Project Manager: A. Hovsepyan, PhD).
  9. SCS MES RA-BRFBR-2014 13РБ-063 “Stimulation of plant growth and development by the use of bacterial melanin synthesized by the insecticidal strain of Bacillus thuringiensis K1”, 2014-2016 (Project Manager: A. Hovsepyan, PhD).
  10. ANSEF No. 3560 “Bacillus thuringiensis strain melanin as a stimulant for plant growth in agricultura”, 2014 (Project Manager: S. Avetisyan, PhD).
  11. 13-2I344 “Construction of recombinant strains possessed by high activity of hydantoinase, isolation and characterization of hydantoinase”, 2013-2015 (Project Manager: A. Hovsepyan, PhD).
  12. 13-2I359 “Cloning of technologically valuable α-amylase and glucoamylase genes and characterization of relevant enzymes”, 2013-2015 (Project Manager: A. Hambardzumyan, PhD).
  13. 11-2i381 “Derivation of strains carrying recombinant plasmids encoding inducable D- and L-aminoacylase and characterization of relevant enzymes”, 2011-2013 (Project Manager: A. Hambardzumyan, PhD).
  14.  ANSEF No. PS-chemorg-2339 “Synthesis of new neuroprotective drugs on the basis of small, substituted carbocycles, preliminary assessment of physiological activities by docking analysis and electrophysiological measurements”, 2011 (Project Manager: A. Mikaelyan).
  15. ISTC #A‑1868 “Development of a new technology for the production of environmentally friendly complex biofertilizers based on nitrogen fixing bacteria and modified zeolites”,  2010-2012 (Project Manager: H. Sargsyan).
  16. ISTC #A‑1866 “Natural biological compounds (peptides) with bactericidal properties against various pathogens”, 2010-2012 (Project Manager: F. Tkhruni, PhD).
  17. ISTC #A-1677 “Synthesis of alpha-amino acids and peptides substituted in a potentially bioactive heterocycles. Study of their antibacterial, antiviral and anti-tumor properties”, 2009-2011 (Project Manager: A. Geolchanyan, PhD).

 

KEY PUBLICATIONS

Scientific articles

  1. Iradyan M., Iradyan N., Hulin Ph., Hambardzumyan A., Gyulkhandanyan A., Alves de Sousa R., Hessani A., Roussakis Ch., Bollot G., Bauvais C. and Sakanyan V. Targeting Degradation of EGFR through the Allosteric Site Leads to Cancer Cell Detachment-Promoted Death Cancers2019, 11, p. 1094. doi:10.3390/cancers11081094.
  2. Hambardzumyan A.A. Statistical analyses of enzyme kinetics: inhibition // Biolog. Journal of Armenia, 2018, 70(1) pp. 35-44.
  3. Aghajanyan A.E., Vardanyan A.A., Hovsepyan A.S., Hambardzumyan A.A., Filipenia V., Saghyan A.S. Development of technology for obtaining water-soluble bacterial melanin and determination of some of pigment properties // BioTechnologia: Journal of Biotechnology, Computational Biology and Bionanotechnology, 2017, 98(4) pp. 315-322. doi.org/10.5114/bta.2017.72292.
  4. Hambardzumyan A.A. Statistical analyses of enzyme kinetics: simple Michaelis-Menten and bi-bi ping-pong // Biolog. Journal of Armenia, 2017, 69(2) pp. 6-12.
  5. Hambardzumyan A.A., Mkhitaryan A.V., Paloyan A.M., and Dadayan S.A. Covalent Immobilization of D-Aminoacylase of Strain Rhodococcus armeniensis AM6.1 and the Characteristics of the Biocatalyst // Applied Biochemistry and Microbiology, 2017, 53(1) pp. 20–24 (Published in Russian in Prikladnaya Biokhimiya i Mikrobiologiya, 2017, Vol. 53, No. 1, pp. 26–30). DOI: 10.1134/S0003683817010057
  6. Hambardzumyan A.A., Mkhitaryan A.V., Paloyan A.M., Dadayan S.A., and Saghyan A.S. Catalytic Properties of Aminoacylase of Strain Rhodococcus Armeniensis AM6.1 // Applied Biochemistry and Microbiology, 2016, 52(3) pp. 250–255 (Original Russian Text: Hambardzumyan A.A., Mkhitaryan A.V., Paloyan A.M., Dadayan S.A., Saghyan A.S. published in Prikladnaya Biokhimiya i Mikrobiologiya, 2016, 52(3) pp. 272–278). DOI: 10.1134/S0003683816030029.
  7. Petrosyan T.R., Ter-Markosyan A.S., Hovsepyan A.S. Detection of Ca2+-dependent acid phosphatase activity identifies neuronal integrity in damaged rat central nervous system after application of bacterial melanin // Neural Regen. Res., 201611(7) pp. 11471152 DOI:10.4103/1673-5374.187055:
  8. Sakanyan V., Hulin P., Alves de Sousa R., Silva V. A. O., Hambardzumyan A., Nedellec S., Tomasoni C., Logé C., Pineau Ch., Roussakis C., Fleury F. & Artaud I. Activation of EGFR by small compounds through coupling the generation of hydrogen peroxide to stable dimerization of Cu/Zn SOD1 // Scientific Reports (Nature Publishing Group), ISSN 2045-2322 (online), 17 February 2016, 6, 21088. doi: 10.1038/srep21088, p. 1-14.
  9. Petrosyan T.R., Gevorgyan O.V. Hovsepyan A.S. and Ter-Markosyan A.S. Effects of bacterial melanin on neuronal activity in the rat sensorimotor cortex // Neurophysiology, 2015, 47(6) pp. 448453. DOI: 10.1007/s11062-016-9554-1.
  10. Gevorkyan O.V., Meliksetyan I.B., Petrosyan T.R., Hovsepyan A.S. Bacterial melanin promotes recovery after sciatic nerve injury in rats // Neural Regen. Res., 2015, 10(1) pp. 124127. DOI:10.4103/1673-5374.150719.
  11. Petrosyan T.R., Gevorkyan O.V., Hovsepyan A.S. Effects of bacterial melanin on movement, posture, and skilled balancing deficits after unilateral destruction of substantia nigra pars compacta in rats // J. Mot. Behav. 2014, 46(1) pp. 6772. DOI: 10.1080/00222895.2013.865588.
  12. Petrosyan T.R., Gevorkyan O.V., Chavushyan V.A., Meliksetyan I.B., Hovsepyan A.S., Manvelyan L.R. Effects of bacterial melanin on motor recovery and regeneration after unilateral destruction of substantia nigra pars compacta in rats // Neuropeptides, 2014, 48(1) pp. 3746. DOI: 10.1016/j.npep.2013.10.001.
  13. Paloyan A.M., Stepanyan L.A., Dadayan S.A., Hambardzumyan A.A., Halebyan Gh.P. and Saghiyan A.S. Catalytic properties of enzymes from Erwinia carotovora involved in transamination of phenylpyruvate // Applied Biochemistry and Microbiology, 2013, 49(2) pp. 106–112 (Original Russian Text: Paloyan A.M., Stepanyan L.A., Dadayan S.A., Hambardzumyan A.A., Halebyan Gh.P., Saghiyan A.S. published in Prikladnaya Biokhimiya I Mikrobiologiya, 2013, 49(2) pp. 129–135). DOI: 10.1134/S0003683813020129.
  14. Paloyan A.M., Hambardzumyan A.A. and Halebyan Gh.P. Isolation, purification, and characterization of phenylpyruvate transaminating enzymes of Erwinia carotovora // Biochemistry (Moscow), 2012, 77(1), pp. 98-104 (Original Russian Text: Paloyan A.M., Hambardzumyan A.A., Halebyan Gh.P. published in Biokhimiya, 2012, 77(1), pp. 123-129). DOI: 10.1134/S0006297912010129.
  15. Petrosyan T.R., Gevorkyan O.V., Meliksetyan I.B., Hovsepyan A.S., Manvelyan L.R. Neuroprotective action of bacterial melanin in rats after corticospinal tract lesions // Pathophysiology, 2012, 19(2) pp. 7180. DOI:10.1016/j.pathophys.2011.12.003.
  16. Aghajanyan A.A., Asaturyan R.A., Hambardzumyan A.A., Sargsyan L.B., Hovsepyan A.S., Vardanyan A.H. and Saghyan A.S. Obtaining of water soluble microbial melanin and study of its some properties // Applied Biochemistry And Microbiology, 2011, 47(5), стр. 500–506 (Original Russian Text: Aghajanyan A.A., Asaturyan R.A., Hambardzumyan A.A., Sargsyan L.B., Hovsepyan A.S., Vardanyan A.H., Saghyan A.S. published in Prikladnaya Biokhimiya i Mikrobiologiya, 2011, 47(5), pp. 551–557). DOI: 10.1134/S0003683811050024.

 

Patents

  1. Paloyan A., Dyukova K., Papoyan A., Aghajanyan A., Hambardzumyan A., Halebyan Gh. Method of obtaining L-phenylalanine // RA Patent No. 2479 A, App. No.  AM20100138, C12P 13/00, 25.01.2011.
  2. Vardanyan A., Aghajanyan A.,  Hovsepyan A., Saghyan A. Microbiological method for obtaining water-soluble melanin // RA Patent No. 3328 A, 2019.

Abstracts Presented at Conferences

1.     Koloyan H.O., Avetisyan S.V., Paronyan M.H., Aganyants H.A., Hovsepyan A.S. Creation of new recombinant strain-producers of l-arginine using shuttle expression vector pEC-XK99E // Book of Abstracts of International Conference “Microbes: Biology & Application, October 9-11, 2019, Yerevan, Armenia, p. 49. ISBN 978-9939-1-1010-3.

2.     Avetisyan S.V., Tadevosyan P.Ye., Koloyan H.O., Paronyan M.H., Hovsepyan A.S. Insecticidal preparations and plant growth stimulator based on melaninogenic Bacillus thuringiensis strains // Book of Abstracts of International Conference “Microbes: Biology & Application, October 9-11, 2019, Yerevan, Armenia, p. 42. ISBN 978-9939-1-1010-3.

3.     Paronyan M., Avetisyan S., Koloyan H., Hovsepyan A., Gyulkhandanyan G. Photodynamic inactivation of methicillin-resistant Staphylococcus aureus and methicillin-sensitive Staphylococcus aureus // Book of Abstracts of International Conference “Microbes: Biology & Application, October 9-11, 2019, Yerevan, Armenia, p. 24. ISBN 978-9939-1-1010-3.

4.     Yepremyan H.S. Approaches to studying the activity and stability of intracellular recombinant aminoacylase of Escherichia coli // Book of Abstracts of International Conference “Microbes: Biology & Application”, October 9-11, 2019, Yerevan, Armenia, p. 14 ISBN 978-9939-1-1010-3.

5.     Hambardzumyan A.A. Application of aminotransferases in asymmetric syntheses of amino acids, Plenary Lecture // Book of Abstracts of International Conference “Microbes: Biology & Application”, October 9-11, 2019, Yerevan, Armenia, p. 27-28. ISBN 978-9939-1-1010-3.

6.     Ghazaryan V.G., Mkhitaryan A.V., Paloyan A.A., Hambardzumyan A.A., Saghyan A.S. Possibility of using thermophilic L-aminoacylases of bacilli origin in the asymmetric syntheses of amino acids // Book of Abstracts of International Conference “Microbes: Biology & Application”, October 9-11, 2019, Yerevan, Armenia, pp. 56-57. ISBN 978-9939-1-1010-3.

7.     Mkhitaryan A.V., Paloyan A.A., Ghazaryan V.G., Hambardzumyan A.A. New Aminoacylase strain from Rhodococcus genus // Book of Abstracts of International Conference “Microbes: Biology & Application”, October 9-11, 2019, Yerevan, Armenia, pp. 58-59. ISBN 978-9939-1-1010-3.

8.     Paloyan A., Melkonyan L., Mkhitaryan A., Dyukova K., Hambardzumyan A. New a-amylase with the potential for commercialization // Book of Abstracts of International Conference “Microbes: Biology & Application”, October 9-11, 2019, Yerevan, Armenia, p. 60. ISBN 978-9939-1-1010-3. (Poster presentation)

9.     Dyukova K.G., Izmailyan M.S., Hambardzumyan A.A. Biosynthesis of extracellular glucose oxidase by Penicillium chrysogenum // Book of Abstracts of International Conference “Microbes: Biology & Application”, October 9-11, 2019, Yerevan, Armenia, pp. 54-55. ISBN 978-9939-1-1010-3.

10.   Aganyants H., Toplaghaltsyan A., Hovhannisyan Ye., Hovsepyan A., Sakanyan V. Influence of phe65ala point mutation on substrate specificity of Geobacillus stearothermophilus d-hydantoinase // Book of Abstracts of International Conference “Microbes: Biology & Application, October 9-11, 2019, Yerevan, Armenia, p. 50. ISBN 978-9939-1-1010-3.

11.   Paloyan A., Melkonyan L., Hambardzumyan A., Mkhitaryan A., Avetisova G. New a-amylase from Bacillus sp. MDC 3500 // Book of Abstracts of International Conference “Designer Biology: From proteins and cells to scaffolds & materials in Europe”, Newcastle University, UK, 31 July - 2 August, 2019, p. 47. (Poster presentation)

12.   Paloyan A., Melkonyan L., Sargsyan A., Arakelyan T., Izmailyan M., Avetisova G., Hambardzumyan A., Sagyan A. Optimization of fermentation condition for a-amylase production // Abstracts submitted to the 43rd FEBS Congress, Prague, Czech Republic, 7-12 July 2018, Poster P. 07-038-Tue, p. 177, FEBS Open Bio 8, Suppl. S1, 2018, pp. 107–496 DOI: 10.1002/2211-5463.12453 177 Є 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.

13.   Melkonyan L., Paloyan A., Hambardzumyan A., Avetisova G., Mkhitaryan A., Sargsyan A., Stepanyan S., Arabachyan S., Saghyan A. Study of some metals effect on a-amylase activity of Bacillus amyloliquefaciens MDC 1974 strain // Abstracts submitted to the 43rd FEBS Congress, Prague, Czech Republic, 7-12 July 2018, 07-042-Wed, p. 178, FEBS Open Bio 8, Suppl. S1, 2018, pp. 107–496 DOI: 10.1002/2211-5463.12453 177 Є 2018 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. (Poster Presentation)

14.   Grigoryan A., Hambardzumyan A., Mikaelyan A. and Melkonyan M. A Comparative Study of Potential Ligands to the Homology Model of Human Alpha2 Adrenoceptor // Abstracts submitted to the 4th Neurological Disorders Summit, Featured Presentation, July 23-25, 2018, Venue Four Points by Sheraton Los Angeles International Airport 9750 Airport Boulevard, Los Angeles, CA 90045, United States, pp. 17-18.

15.   Grigoryan A., Babayan B., Melkonyan M., Hambardzumyan A. and Mikaelyan A. Identification of Binding Affinities of Some Pyrethroid-Like Cyclopropane/Cyclobutane Ligands with VG SCN and NMDA Receptor // Abstracts submitted to the 4th Neurological Disorders Summit, July 23-25, 2018, Venue Four Points by Sheraton Los Angeles International Airport 9750 Airport Boulevard, Los Angeles, CA 90045, United States, pp. 75-76. (Poster Presentation)

16.   Grigoryan A., Babayan B., Avetisyan S., Sargsyan V., Hambardzumyan A., Mikaelyan A. Comparative docking analysis of Cyclopropane/Cyclobutane derivatives with human modeled and bacterial voltage gated sodium channels and NMDA receptor // Book of Abstracts of the 1st International Scientific Conference “Current State of Pharmacy and Prospects of its Development”, November 01-03, 2018, Yerevan, Armenia, pp. 30-31. (Report)

17.   Aganyanc H., Hovhannisyan Y., Toplaghaltsyan A., Hambardzumyan A., Hovsepyan A., Sakanyan V. Approaches for obtaining new highly active recombinant thermostable D-hydantoinases // Book of Abstracts of 1st International Scientific Conference “Current State of Pharmacy and Prospects of its Development”, November 01-03, 2018, Yerevan, Armenia, p. 89. (Poster Presentation)

 

Staff

Anichka

Hovsepyan

Leading Researcher,

PhD in Biology

anichkahovsepyan@armbiotech.am  

Hasmik

Yepremyan

Senior Researcher,

PhD in Chemistry

 

Haykanush

Koloyan

Senior Researcher,

PhD in Biology

ankoloyan@armbiotech.am 

Karine

Dyukova

Senior Researcher,

PhD in Chemistry

karinedyukova@armbiotech.am 

Ani

Paloyan

Senior Researcher,

PhD in Biology

 anipaloyan@armbiotech.am

Sona

Avetisyan

Senior Researcher,

PhD in biology

 sonaavetisyan@armbiotech.am

Marina

Paronyan

Researcher,

PhD in Biology

 marinaparonyan@armbiotech.am 

Anna

Mkhitaryan

Junior Researcher,

PhD in Biology

 annamkhitaryan@armbiotech.am

Marieta

Izmailyan

 

Junior Researcher

 

Narine

Mikayelyan

 

Laboratory Assistant