Dr. Paula Adriana Pitashny

Dr. Paula Adriana Pitashny
Principal Investigator
Principal Investigator
Senior Lecturer
PhD, 1998, Technion, Israel Institute of Technology
Medicinal Cannabis
Phone
972-54-6642014
Research interests:

Our research group is interested in the investigation of the potential medicinal activities of the different cannabis plant species in pre-clinical studies. We are investigating the possibility that cannabis species and their active compounds will be able to ameliorate the neurological disorders and symptoms of the Rett Syndrome using Zebra Fish Larvae model. Cannabis extracts are being investigated for pain relieve, appetite, anxiety and sleep disorders. The effects of cannabis on the different symptoms are investigated at the molecular level by RT-PCR. In addition, we also investigate the effects of cannabis on inflammatory diseases as well as cancer, on cells and animal models. All the results obtained by pre-clinical studies are correlated with chemical profiles of the cannabis species. 

White Rot Fungi, Peroxidases, Melanin Degradation, Medicinal Cannabis, Zebra Fish Model

2004- Present. Senior Lecturer at Tel Hai College, Upper Galilee, Israel.

 

Administrative positions in academic institutions

2018-Present Head of Food Science Department, Faculty of science and Technology at Tel Hai College, Upper Galilee, Israel.

2007-2013 Head of Food Science Department, Faculty of science and Technology at Tel Hai College, Upper Galilee, Israel.

 

  1. Positions in non academic & research Institutions

2018-Present   Coordinator of Medicinal Cannabis Center at Migal-Galilee Research Institute. Kiriat Shmona, Israel.

2002-Present  Chief Technology Officer (CTO) and R&D Director at Rakuto Biotechnologies Ltd.-Syneron-Candela Ltd, Yokneam, Israel.

2000-2013     Head of Environmental Biotechnology Lab at Migal-Galilee Research Institute. Kiriat Shmona, Israel.

 

  1. Membership in professional organizations  

         2009 The American Society of Microbiology

 

  1. Research Grants

2003   Israel Science Foundation, The Academy of Sciences and Humanities (for 3 years).

2001  MOP Ezori Grant awarded by the Ministry of Culture, Sciences and Sport, Israel Humanities (for 3 years).

2000    The Guastella fellowship, for the academic years 2000-2002 was awarded by the Sacta-Rashi Foundation, Jerusalem, Israel.

 

  1. Teaching at Academic institutions

2009-2014 Lecturer

Department of Biotechnology, Faculty of Science and Technology (M.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: Bioreactors.

 

2007-2012 Lecturer

Department of Biotechnology, Faculty of Science and Technology (B.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: Advanced Clinical Molecular Biology.

 

2006-Present Lecturer

Department of Nutritional Sciences and Department of Food Technology, Faculty of Science and Technology (B.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: Advanced Seminar.

 

1999-Present Lecturer

Department of Biotechnology, Department of Environmental Sciences, Department of Nutritional Sciences, Department of Food Sciences and Department of Zoo-technology, Faculty of Science and Technology (B.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: General and Inorganic Chemistry.

 

1998-2014 Lecturer

Department of Biotechnology, Department of Nutritional Sciences, Department of Food Sciences, Faculty of Science and Technology (B.Sc.). Tel Hai College, Upper Galilee, Israel.

Course: Antioxidants in Medicine and Food.

CV
CURRICULUM VITAE

Paula Adriana Pitashny (Belinky)

 

  1. Personal Details:

Date of birth: 9/8/1962

Country of birth: Argentina

Israelian Identity No.: 01317959-3

Argentinian D.N.I.: 14830853

Nationality: Argentina, Israel

Family status: Divorced + 2

Permanent Address: Ha Narkis 25, Metulla, 10292, Israel

Phone numbers: Home- +972-4-6339109; Mobile- +972-54-6642014

E-mail address: paula@migal.org.il

 

  1. Higher Education

1981-1985 B.Sc. Studies in Chemistry, Faculty of Natural Sciences, Hebrew University of Jerusalem, Israel.

 

    1. M.Sc. Department of Biochemistry and Human Nutrition, Faculty of Agriculture, The Hebrew University of Jerusalem, Israel. Supervisors: Prof. Ilan Het, Prof. Ytzhak Hadar and Prof. Dan Lebanon. Thesis: Biosynthesis of the flavor compound 1-octen-3-ol by the edible mushroom Pleurotus pulmonarius in submerged fermentation.

 

    1.  Ph.D. Department of Lipids, Faculty of Medicine, Technion, Israel Institute of Technology. Supervisors: Prof. Michael Aviram and Prof. Jacob Vaya. Thesis: Effect of antioxidant constituents from the roots of Glycyrrhiza glabra  (licorice) on the oxidation of low density lipoprotein (LDL).

 

    1. Post Doc. Department of Immunology, Migal, Galilee Research Institute, Israel. RNA-trans splicing. Prof. Gideon Gross.
  1. Appointments at academic institutions

2014-2015 Sabbatical year in South America                     

2004- Present. Senior Lecturer at Tel Hai College, Upper Galilee, Israel.

 

  1. Administrative positions in academic institutions

2018-Present Head of Food Science Department, Faculty of science and Technology at Tel Hai College, Upper Galilee, Israel.

2007-2013 Head of Food Science Department, Faculty of science and Technology at Tel Hai College, Upper Galilee, Israel.

 

  1. Positions in non academic & research Institutions

2018-Present   Coordinator of Medicinal Cannabis Center at Migal-Galilee Research Institute. Kiriat Shmona, Israel.

2002-Present  Chief Technology Officer (CTO) and R&D Director at Rakuto Biotechnologies Ltd.-Syneron-Candela Ltd, Yokneam, Israel.

2000-2013     Head of Environmental Biotechnology Lab at Migal-Galilee Research Institute. Kiriat Shmona, Israel.

 

  1. Membership in professional organizations  

         2009 The American Society of Microbiology

 

  1. Research Grants

2003   Israel Science Foundation, The Academy of Sciences and Humanities (for 3 years).

2001  MOP Ezori Grant awarded by the Ministry of Culture, Sciences and Sport, Israel Humanities (for 3 years).

2000    The Guastella fellowship, for the academic years 2000-2002 was awarded by the Sacta-Rashi Foundation, Jerusalem, Israel.

 

  1. Teaching at Academic institutions

2009-2014 Lecturer

Department of Biotechnology, Faculty of Science and Technology (M.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: Bioreactors.

 

2007-2012 Lecturer

Department of Biotechnology, Faculty of Science and Technology (B.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: Advanced Clinical Molecular Biology.

 

2006-Present Lecturer

Department of Nutritional Sciences and Department of Food Technology, Faculty of Science and Technology (B.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: Advanced Seminar.

 

1999-Present Lecturer

Department of Biotechnology, Department of Environmental Sciences, Department of Nutritional Sciences, Department of Food Sciences and Department of Zoo-technology, Faculty of Science and Technology (B.Sc.), Tel Hai College, Upper Galilee, Israel.

Course: General and Inorganic Chemistry.

 

1998-2014 Lecturer

Department of Biotechnology, Department of Nutritional Sciences, Department of Food Sciences, Faculty of Science and Technology (B.Sc.). Tel Hai College, Upper Galilee, Israel.

Course: Antioxidants in Medicine and Food.

 

1993-1998 Teaching Assistant.

Department of Biotechnology and Environmental Sciences, Faculty of Science and Technology (B.Sc.) and Faculty of Agriculture of the Hebrew University of Jerusalem, Tel Hai College, Upper Galilee, Israel.

Course: Organic Chemistry.

 

1989-1993  Teaching Assistant.

Tel Hai College, Upper Galilee, Israel.

Course: Laboratory of Microbiology.

  1. Supervision of Master’s and doctoral degree students

2012-2015 Mayan Sair. Influence of nutrients in the production of linin peroxidase by Phanerochaete chrysosporium. Master Degree at Tel Hai College.

2011-2013 Aya Sitruk. Sources of hydrogen peroxide in Phanerochaete chrysosporium. Master Degree at Tel Hai College.

2009-2011 Shaul Barkaly. Homologous expression of lignin peroxidase in yeasts. Master Degree at Tel Hai College.

2007-2011  Meytal Nuriel. Characterization of manganese superoxide dismutase lacking mutants of Phanerochaete chrysosporium. Master Degree at Faculty of Agriculture, Food and Environment, Hebrew University in Jerusalem, Hebrew University in Jerusalem in cooperation with Prof. Yitzhak Hadar.

2005-2011 Avi Matityahu. Role of reactive oxygen species in the induction of lignin peroxidase expression in the white-rot fungus Phanerochaete chrysosporium. Doctor Degree at Faculty of Agriculture, Food and Environment, Hebrew University in Jerusalem, in cooperation with Prof. Yitzhak Hadar.

2004-2007 Shaul Cohen. Oxidation of catechols by lignin peroxidase. Master Degree at Faculty of Agriculture, Food and Environment, Hebrew University in Jerusalem, in cooperation with Prof. Ytzhak Hadar.

2000-2002 Nufar Flikshtein. The role of oxygen in the induction of synthesis of lignin peroxidase (LIP) in white rot fungi Phanerochaete chrysosporium. Master Degree at Technion, Israel Institute of Technology in cooperation with Prof. Shimon Gepstein and Prof. Carlos Dosoretz.

 

  1. List of Publications
  1. Belinky, P. A.; Masaphy, S.; Lebanon, D.; Hadar, Y.; Dosoretz, C. G. Effect of medium composition on 1-octen-3-ol formation in submerged cultures of Pleurotus pulmonarius. Appl. Microbiol. Biotechnol. 40:633-640; 1993.
  2.  Vaya, J.; Belinky, P. A.; Aviram, M. Antioxidants constituents from licorice roots: isolation, structure elucidation and antioxidative capacity towards LDL oxidation. Free Radic. Biol. Med. 23:302-313; 1997.
  3. Fuhrman, B.; Buch, S.; Vaya, J.; Belinky, P. A.; Coleman, R.; Hayek, T.; Aviram, M. Licorice extract and its major polyphenol glabridin protect low-density lipoprotein against lipid peroxidation: in vitro an ex-vivo studies in humans and in the atherosclerotic apolipoprotein E deficient mice. Am.  J. Clin. Nutr. 66:267-275; 1997.
  4. Hayek, T.; Fuhrman, B.; Vaya, J.; Rosenblat, M.; Belinky, P. A.; Coleman, R.; Elis, A.; Aviram, M. Reduced progression of atherosclerosis in apolipoprotein E deficient mice following consumption of red wine, or its polyphenols quercetin or catechin, is associated with reduced susceptibility of LDL to oxidation and aggregation. Arterioscler. Thromb. Vasc. Biol. 17:2744-2752; 1997.
  5.  Belinky, P. A.; Aviram, M.; Fuhrman, B.; Rosenblat, M.; Vaya, J. The antioxidative effects of the isoflavan glabridin on endogenous constituents of LDL during its oxidation. Atherosclerosis 137:49-61; 1998.
  6. Belinky, P. A.; Aviram, M.; Mahmood, S.; Vaya, J. Structural aspects of the inhibitory effect of glabridin on LDL oxidation. Free Radic. Biol. Med. 24:1419-1429; 1998.
  7. Rosenblat, M.; Belinky, P. A.; Vaya, J.; Levy, R.; Merchav, S.; Aviram, M. Macrophage enrichment with the isoflavan glabridin inhibits NADPH oxidase-induced cell-mediated oxidation of low density lipoprotein (LDL): a possible role for protein kinase C. J. Biol. Chem. 274:13790-13799; 1999.
  8. Fuhrman, B., Vaya, J., Belinky, P. A., Aviram, M. The isoflavan glabridin inhibits LDL oxidation : structure and mechanistic aspects. (Natural antioxidants and anticarcinogens in nutrition, health and disease) Spec. Publ.-R. Soc. Chem. 240:161-165; 1999.  
  9. Ward, G.; Belinky, P. A.; Hadar, Y.; Bilkis, I.; Dosoretz, C. G. The influence of non-phenolic mediators and phenolic co-substrates on the oxidation of 4-bromophenol by lignin peroxidase. Enzyme Microb. Technol. 30:490-498; 2002.
  10.  Belinky, P. A.; Goldberg, D.; Krinfeld, B.; Burger, M.; Rothschild, N.; Cogan, U.; Dosoretz, C. G. Manganese-containing superoxide dismutase from the white-rot fungus Phanerochaete chrysosporium: its function, expression and gene structure. Enzyme Microb. Technol. 31:754-764; 2002.
  11.   Belinky, P. A.; Flikshtein, N.; Lechenko, S.; Gepstein, S.; Dosoretz, C. G. Reactive oxygen species and the induction of lignin peroxidase in Phanerochaete chrysosporium. Appl. Environ. Microbiol. 69:6500-6506; 2003.
  12.  Belinky, P. A; and Dosoretz, C. G. ROS induction of LIP gene expression. Free Rad. Res. Proceedings of the Meeting of the Society for Free Radicals-SFRR Europe. Free radicals and oxidative stress: chemistry, biochemistry and pathophysiological implications. 31-34; 2003.
  13.  Schlesinger, J.; Arama, D.; Noy, H.; Dagash, M; Belinky, P. A.; Gross, G.  In-cell joining of antibody variable region genes and generation of single chain Fv transcripts via targeted RNA trans-splicing. J. Immunol. Meth. 282:175-86; 2003.
  14. Belinky , P. A.; Flicktein, N; Dosoretz, C. G. Induction of lignin peroxidase via reactive oxygen species in manganese-deficient cultures of Phanerochaete chrysosporium. Enzyme Microbiol. Technol. 39:222-228; 2006.
  15. Wymelenberg, A. V.; Minges, P.; Sabat, G.; Martinez, D.; Aerts, A.; Salamov, A.; Grigoriev, I.; Shapiro,H.; Nik Putman, N.; Belinky, P. A.; Dosoretz, C. G.; Gaskell, J.; Phil Kersten, P.; Cullen, D. Computational analysis of the Phanerochaete chrysosporium v2.0 genome database and mass spectrometry identification of peptides in ligninolytic cultures reveal complex mixtures of secreted proteins. Fungal Gen. and Biol. 43:343-356; 2006.
  16. Matityahu, A.; Hadar, Y.; Dosoretz, C. G; Belinky, P. A. Gene Silencing by RNA Interference in the White-Rot Fungus Phanerochaete chrysosporium. Appl. Environ. Microbiol. 74:5359-5365; 2008.
  17. Cohen, S.; Belinky, P. A.; Hadar, Y.; Dosoretz, C. G. Characterization of catechol derivative removal by lignin peroxidase (LIP) in aqueous mixture. Bioresource Technol. 100:2247-2253; 2009.
  18. Matityahu, A, Hadar, Y, Belinky, P.A. Involvement of Reactive Oxygen Species and Protein Kinase C in Lignin Peroxidase Expression in Oxygenated Cultures of the White Rot Fungus Phanerochaete chrysosporium. Enzyme Microbiol. Technol. 47:59-63; 2011.
  19. Matityahu, A, Hadar, Y, Sitruk, A., Belinky, P. A. Factors affecting the Induction of Lignin Peroxidase in Manganese Deficient Cultures of the White Rot Fungus Phanerochaete chrysosporium. Adv. Microbiol. 5:83-92; 2015.

 

 

Articles in preparation:

Sitruk, A., Matityahu, A., Belinky, P. A. Sources for production of hydrogen peroxide in Phanerochaete chrysosporium. In preparation.

Sitruk, A., Matityahu, A., Belinky, P. A. Cultures conditions affecting lignin peroxidase by the white rot fungus Phanerochaete chrysosporium. In preparation.

Barkly, S., Matityahu, A., Belinky, P. A. Homologous expression of lignin peroxidase in Picchia pastoris. In preparation.

Nuriel, M., Matityahu, A., Belinky, P. A. Characterization of MnSOD lacking mutants of Phanerochaete chrysosporium in relation to the production of lignin peroxidase. In preparation.

 

  1. Patents

1. Belinky, P. A.; Lasser, H; Dosoretz, C. G. Methods of producing lignin peroxidase and its use in skin and hair lightening. Patent number: US 7,422,734 B2. 2008.

2. Belinky, P. A.; Karmon, Y.; Krinfeld, B.; Lasser, H. Enzymatic system-containing cosmetic compositions. 56558 PCT: 58615| US. Provisional Application No. 61/827,685 filed 27-May-2013.  

 

  1. Ph.D./ Master Dissertation

M.Sc. Thesis

Belinky, P. A. “Biosynthesis of the flavor compound 1-octen-3-ol by the edible mushroom Pleurotus pulmonarius in submerged fermentation”. Department of Biochemistry and Human Nutrition, Faculty of Agriculture, Rehovot, The Hebrew University of Jerusalem. (M.Sc. research thesis [Hebrew], submitted on 1991).

 

Ph.D. Thesis

Belinky, P. A. “Effect of antioxidant constituents from the roots of Glycyrrhiza glabra  (licorice) on the oxidation of low density lipoprotein (LDL)”. Department of Lipids, Faculty of Medicine, Technion, Israel Institute of Technology. (Ph.D. research thesis [English], submitted on 1998).

  

  1. Participation in Scientific Conferences

2020    The international Medical Cannabis Conference, Lisbon, Portugal. Pitashny, P.A.; Harari, R.; Jakson, K. Classification of medical cannabis strains for treatment of pain by zebrafish model. Poster presentation.

 

2019    First Congress of the Israel Association of Cannabis Medicine, Tel Aviv.

2019  The second International Annual Congress of Medicinal Cannabis Controversies. Barcelona, Spain

2012 Gordon Research Conferences. Cellular & Molecular Fungal Biology. Holderness, New Hamshire, USA. Matityahu, A.; Belinky, P. A. Involvement of protein kinase C in lignin peroxidase expression in oxygenated cultures of the white rot fungus Phanerochaete chrysosporium. Poster presentation.

2008 9th European Conference on Fungal Genetics University of Edinburgh, Scotland. Matityahu, A.; Hadar, Y.; Belinky, P. A. Gene Silencing by RNA Interference in the White Rot Fungus Phanerochaete chrysosporium. Poster presentation.

          2008 The 5th Congress of the Federation of Israel Societies for Experimental Biology, Eilat, Israel. Matityahu, A.; Belinky, P. A. The role of ROS and MnSOD in the induction of Lignin Peroxidase in Phanerochaete chrysosporium. Poster presentation.

2007  23rd Annual Meeting of the Israel Society for oxygen and free radical research. Tel Aviv University, Israel. Matityahu, A.; Belinky, P. A. Induction of Lignin Peroxidase in Phanerochaete chrysosporium. Poster presentation.

2007 23rd Annual Meeting of the Israel Society for oxygen and free radical research. Tel Aviv University, Israel. Cohen, S.; Belinky, P.A. Oxidation of Catechols by Lignin Peroxidase. Poster presentation.

2006 22nd Annual Meeting of the Israel Society for oxygen and free radical research. Tel Aviv University, Israel. Matityahu, A.; Belinky, P.A. Reactive Oxygen Species and Induction of Lignin Peroxidase in Phanerochaete chrysosporium. Poster presentation.

2005 The 4th Congress of the Federation of Israel Societies for Experimental Biology, Eilat, Israel. Matitiahu, A.; Belinky, P.A. Reactive Oxygen Species and Induction of  Lignin Peroxidase in Phanerochaete chrysosporium. Poster presentation. Cohen, S.; Belinky, P.A. Oxidation of Catechol by lignin peroxidase. Poster presentation.

  1. Society for free radical research SFRR-Europe meeting 2003: “Free radicals and Oxidative Stress: Chemistry Biochemistry and Pathophysiological implications”, Ioannina, Greece. Belinky, P.A.; N. Flikshtein, N.;  Lechenko, S.; Gepstein, S and Dosoretz, C.G. Involvement of Reactive Oxygen Species in the induction of lignin peroxidase gene expression in liquid cultures of Phanerochaete chrysosporium. Lecture.

2002 Annual meeting of the Israel Society of Microbiology, Hebrew University, Jerusalem, Israel. Flikshtein, N.; Belinky, P.A;  Lechenko, S.; Gepstein, S and Dosoretz, C.G. Effect of molecular oxygen in liquid cultures of Phanerochaete chrysosporium. Poster presentation.

2002 The 3rd Congress of the Federation of Israel Societies for Experimental Biology, Eilat, Israel. Belinky, P.A.; N. Flikshtein, N.; Lechenko, S.; Gepstein, S and Dosoretz, C.G. Effect of molecular oxygen in liquid cultures of Phanerochaete chrysosporium. Poster presentation.

1998 Meeting of the Society for Research, Prevention and Treatment of Atherosclerosis, Ashkelon, Israel. Mahmood, S.; Belinky, P. A.; Tamir, Snait; Vaya, J. Quantum Chemical Computation of glabridin in relation to its inhibitory effect on LDL oxidation. Poster presentation.

1998 The 70th meeting of the European Atherosclerosis Society, Geneve, Switzerland. Belinky, P. A.; Aviram, M.; Mahmood, S.; Vaya, J. Structural aspects of the inhibitory effect of glabridin on LDL oxidation.  Poster presentation.

1998 The first Regional Meeting on Medical Sciences: The Role of Free Radicals in Health and Disease, Jerusalem and Amman, Jordan. Belinky, P. A.; Aviram, M.; Mahmood, S.; Vaya, J. Structural aspects of the inhibitory effect of glabridin on LDL oxidation. Poster presentation.

1998 The 4th Annual Meeting of the Society for Research, Prevention and Treatment of Atherosclerosis, Eilat, Israel. Belinky, P. A.; Aviram, M.; Fuhrman, B.; Rosenblat, M.; Vaya, J. The antioxidative effects of the isoflavan glabridin on endogenous constituents of LDL during its oxidation. Poster presentation.

1997 The Annual Meeting of the Israeli Society of Free Radicals, Bar Ilan University, Israel. Belinky, P. A.; Aviram, M.; Fuhrman, B.; Rosenblat, M.; Vaya, J. The antioxidative effects of the isoflavan glabridin on endogenous constituents of LDL during its oxidation. Lecture.

 

  1. Research

1) Lignin peroxidase expression in the white rot fungus Phanerochaete chrysosporium

The most intensively studied white rot fungus, Phanerochaete chrysosporium, secretes an array of peroxidases which are able to extensively degrade lignin, the most aboundant aromatic polymer. Two major families of hydrogen peroxide (H2O2)-requiring extracellular heme-peroxidases designated lignin peroxidase (LIP) and manganese-dependent peroxidase (MNP) are secreted. The non-specific nature and exceptional oxidation potential of the lignin peroxidases has attracted considerable interest in organopollutants degradation and fiber bleaching. The regulation of the gene families encoding extracellular peroxidases is poorly understood, but it’s clear that oxygen stress is a key factor. LIP expression requires nutrient or carbon starvation, signaled by a rise in intracellular cAMP concentrations and concomitant pulses of pure oxygen gas in the headspace. The effect of such elevated oxygen concentrations on LIP expression can be reproduced by depleting Mn2+ ions from cultures, in the presence of atmospheric air. Development of a putative oxidative stress is detected in LIP-producing cultures of P. chrysosporium, either oxygenated or Mn2+-deficient. In oxygenated cultures, increased expression of MnSOD was the major response of the antioxidant system. In contrast, in Mn2+-deficient cultures, negligible activity of MnSOD was detected.

Addition of a OH scavenger, dimethylsulfoxide, to the cultures completely abolished LIP transcription (both mRNA and heme-protein were undetectable), indicating that these ROS, coupled to high levels of cAMP, are indeed involved in the induction of LIP expression. Since the difference between both types of LIP-producing cultures consist in activation or lack of MnSOD activity, it is possible that ROS production in general and OH generation in particular occurs by different mechanisms. The long-term objective of this research is to study the role of ROS in the regulation or induction of LIP expression in the white-rot fungus P. chrysosporium as well as the sources, ways and mechanisms of their generation.

The construction of conditional MnSOD(-) mutants will enable us to ascertain the role or influence of MnSOD, Mn2+ ions as well as other antioxidant enzymes in the production of the relevant ROS necessary for LIP induction. The results of this research are expected to advance the understanding of the pathways, sources and nature of ROS as second messengers in LIP expression.

Work program:

1- Effect of reactive oxygen expression (ROS) on lignin peroxidase expression in the white rot fungus P. chrysososporium.

It was found that hydroxyl radical is the principal ROS for enhancement of lignin peroxidase expression. The sources of this radical in different lignin peroxidase producing cultures are under investigation.

2- Role of manganese superoxide dismutase in lignin peroxidase expression.

Mutants lacking manganese superoxide dismutase were prepared by RNA silencing. Those mutants will be characterized and they will be used for understanding the role of manganese ions and manganese superoxide dismutase in lignin peroxidase expression.

3- Signal transduction pathway and lignin peroxidase expression.

The relationship between protein kinase C and lignin peroxidase will be investigated.

4- Improvement of lignin peroxidase production by P. chrysosoporium.

Optimization of lignin peroxidase production by changing different growth parameters and medium composition will be performed.

 

2) Homologous expression of lignin peroxidase in yeasts.

Potential applicability of LIP depends on the ability to produce high quantities of the enzyme by efficient growth and purification technologies.

Lignin peroxidase production by submerged fermentation of P. chrysosporium is hampered by several factors, such as expression under nutrient limitation together with oxidative stress, and in the sensitivity of this basidiomycete fungus to high shear forces in a fermenter. Moreover, the purification stage of the enzyme from the growth liquid medium of P. chrysosporium is tedious and expensive.

Homologous expression of LIP in yeasts can be an alternative and more efficient process for LIP production, since less incubation time is needed for the organism growth and only LIP-H2 could be expressed, avoiding the purification step from other isoenzymes. 

The present study addresses the production of lignin peroxidase H2 in the yeast Pichia pastoris.

Work program

1- Cloning of the encoding region of LIP-H2 into the expression vector pPIC9 from the expression system of P. pastoris (invitrogene).

2- Transformation to P. pastoris.

3- Induction of LIP enhanced expression, collection and purification of the secreted enzyme

4- Structural and activity comparison between the enzyme expressed by P. pastoris and the enzyme produced by P. chrysosporium

3) Large scale lignin peroxidase production

Development and scale up for production of lignin peroxidase by different white rot fungi.

Scale up of downstream process.

Optimization of spore production at large quantities.

 

4) Development of formulations containing lignin peroxidase for skin whitening

Test of compatibility of the different formula components with the enzyme.

Determination of optimal melanin oxidation by lignin peroxidase formulation.

 

5) Effect of Cannabis extracts on Zebra Fish behavior.

 

  1. Referees  

Prof. Carlos G. Dosoretz. Technion, Israel Institute of Technology. Civil Environmental Engineering Faculty. Tel.: +972-4-8294962, Fax.: +972-4-8228898, e-Mail: carlosd@tx.technion.ac.il

Prof. Yitzhak Hadar. Faculty of Agriculture, Food and Environment, Hebrew University of Jerusalem, Plant Pathology and Microbiology Department. Tel.: +972-8-9489935, Fax: +972-8-9468785, E-mail: hadar@agri.huji.ac.il

Prof. Michael Aviram. Technion, Israel Institute of Technology, Faculty of Medicine. Tel.: +972-4-8295278. e-Mail: aviram@tx.technion.ac.il      

             

 

 

Funding

2003   Israel Science Foundation, The Academy of Sciences and Humanities (for 3 years).

2001  MOP Ezori Grant awarded by the Ministry of Culture, Sciences and Sport, Israel Humanities (for 3 years).

2000    The Guastella fellowship, for the academic years 2000-2002 was awarded by the Sacta-Rashi Foundation, Jerusalem, Israel.

Scientific Publications

Factors affecting the Induction of Lignin Peroxidase in Manganese Deficient Cultures of the White Rot Fungus Phanerochaete chrysosporium.

Matityahu, A, Hadar, Y, Sitruk, A., Belinky, P. A.
2015

Involvement of Reactive Oxygen Species and Protein Kinase C in Lignin Peroxidase Expression in Oxygenated Cultures of the White Rot Fungus Phanerochaete chrysosporium.

Matityahu, A, Hadar, Y, Belinky, P.A.
2011

Characterization of catechol derivative removal by lignin peroxidase (LIP) in aqueous mixture.

Cohen, S.; Belinky, P. A.; Hadar, Y.; Dosoretz, C. G.
2009

Gene Silencing by RNA Interference in the White-Rot Fungus Phanerochaete chrysosporium.

Matityahu, A.; Hadar, Y.; Dosoretz, C. G; Belinky, P. A.
2008

Induction of lignin peroxidase via reactive oxygen species in manganese-deficient cultures of Phanerochaete chrysosporium.

Belinky , P. A.; Flicktein, N; Dosoretz, C. G.
2006

Computational analysis of the Phanerochaete chrysosporium v2.0 genome database and mass spectrometry identification of peptides in ligninolytic cultures reveal complex mixtures of secreted proteins.

Wymelenberg, A. V.; Minges, P.; Sabat, G.; Martinez, D.; Aerts, A.; Salamov, A.; Grigoriev, I.; Shapiro,H.; Nik Putman, N.; Belinky, P. A.; Dosoretz, C. G.; Gaskell, J.; Phil Kersten, P.; Cullen, D.
2006

Reactive oxygen species and the induction of lignin peroxidase in Phanerochaete chrysosporium.

Belinky, P. A.; Flikshtein, N.; Lechenko, S.; Gepstein, S.; Dosoretz, C. G.
2003

ROS induction of LIP gene expression.

Belinky, P. A; and Dosoretz, C. G.
2003

In-cell joining of antibody variable region genes and generation of single chain Fv transcripts via targeted RNA trans-splicing.

Schlesinger, J.; Arama, D.; Noy, H.; Dagash, M; Belinky, P. A.; Gross, G.
2003

The influence of non-phenolic mediators and phenolic co-substrates on the oxidation of 4-bromophenol by lignin peroxidase.

Ward, G.; Belinky, P. A.; Hadar, Y.; Bilkis, I.; Dosoretz, C. G.
2002

Manganese-containing superoxide dismutase from the white-rot fungus Phanerochaete chrysosporium: its function, expression and gene structure.

Belinky, P. A.; Goldberg, D.; Krinfeld, B.; Burger, M.; Rothschild, N.; Cogan, U.; Dosoretz, C. G.
2002

Macrophage enrichment with the isoflavan glabridin inhibits NADPH oxidase-induced cell-mediated oxidation of low density lipoprotein (LDL): a possible role for protein kinase C.

Rosenblat, M.; Belinky, P. A.; Vaya, J.; Levy, R.; Merchav, S.; Aviram, M.
1999

The isoflavan glabridin inhibits LDL oxidation : structure and mechanistic aspects. (Natural antioxidants and anticarcinogens in nutrition, health and disease)

Fuhrman, B., Vaya, J., Belinky, P. A., Aviram, M.
1999

The antioxidative effects of the isoflavan glabridin on endogenous constituents of LDL during its oxidation.

Belinky, P. A.; Aviram, M.; Fuhrman, B.; Rosenblat, M.; Vaya, J.
1998

Vaya, J. Structural aspects of the inhibitory effect of glabridin on LDL oxidation.

Belinky, P. A.; Aviram, M.; Mahmood, S.; Vaya, J.
1998

Licorice extract and its major polyphenol glabridin protect low-density lipoprotein against lipid peroxidation: in vitro an ex-vivo studies in humans and in the atherosclerotic apolipoprotein E deficient mice.

uhrman, B.; Buch, S.; Vaya, J.; Belinky, P. A.; Coleman, R.; Hayek, T.; Aviram, M.
1997

Reduced progression of atherosclerosis in apolipoprotein E deficient mice following consumption of red wine, or its polyphenols quercetin or catechin, is associated with reduced susceptibility of LDL to oxidation and aggregation.

Hayek, T.; Fuhrman, B.; Vaya, J.; Rosenblat, M.; Belinky, P. A.; Coleman, R.; Elis, A.; Aviram, M.
1997

Antioxidants constituents from licorice roots: isolation, structure elucidation and antioxidative capacity towards LDL oxidation.

Vaya, J.; Belinky, P. A.; Aviram, M.
1997

Effect of medium composition on 1-octen-3-ol formation in submerged cultures of Pleurotus pulmonarius.

Belinky, P. A.; Masaphy, S.; Lebanon, D.; Hadar, Y.; Dosoretz, C. G.
1994