Review
BibTex RIS Cite

PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN

Year 2017, Volume: 6 Issue: 2, 103 - 113, 31.08.2017
https://doi.org/10.18036/aubtdc.307261

Abstract

Bacillus thuringiensis (Bt) is a gram positive and spore forming bacterium that shows a wide distribution in the nature.  During the sporulation phase, it produces insecticidal crystal proteins along with spores. These insecticidal proteins are called Cry and Cyt proteins and have an activity when binding to spesific receptor located on the plasma membrane of mid-gut epithelial cells of susceptible insects. This receptor mediated spesific activity makes Bt important in agricultural control of insects. Afterwards, research on non-insecticidal proteins that have a wider distribution in nature than insecticidal ones revealed a new protein class called parasporin. Parasporins are capable of discriminately killing cancer cells while does not show cytotoxic and hemolytic activity against normal cells. So far, six parasporin families have been defined and a total of 19 sub-group is known.  Because  parasporins have a similar spesific-receptor binding effect with insecticidal crystal proteins,  they had led to emergence of a new research field related to cancer treatment.  This study reviewed the discovery of parasporins, their characteristics, varieties and their mechanisms of action on cancer cells.

References

  • KAYNAKLAR [1] Ishiwata S. On a kind of severe flacherie (sotto disease). Dainihon Sanshi Kaiho 1901; 114: 1-5.
  • [2] Ben- Dov E., Zaritsky A., Dahan E., Barak Z., Sınal R., Manasherob R., Khamraev A., Troitskaya E., Dubitsky A., Berezina N., Margalith Y. Extended screening by PCR for seven cry-group genes from field collected strains of Bacillus thuringienis. Appl Environ Microbiol 1997; 63: 4883- 4890.
  • [3] Bernhard K., Jarrett P., Meadows M., Butt J., Ellis D.J., Roberts G.M., Pauli S., Rodgers P., Burges H.D. Natural Isolates Of Bacillus Thuringiensis: Worldwide Distribution, Characterization, And Activity Against Insect Pests. J Invertebr Pathol 1997; 70 (1): 59-68.
  • [4] Ichimatsu T., Mizuki E., Nishimura K., Akao T., Saitoh H., Higuchi K., Ohba M. Occurrence of Bacillus thuringiensis in fresh water of Japan. Curr Microbiol 2000; 40: 212- 217.
  • [5] Logan N. Bacillus anthracis, Bacillus cereus, and other aerobic endospore-forming bacteria. Topley & Wilson’s Microbiology & Microbial Infections (Tenth Edition). Bacteriology. Borriello SP, Murray PR and Funke G (eds.). London: Hodder Arnold; 922-952, 2005.
  • [6] Bravo A., Gill S.S., Soberon M. Bacillus thuringiensis mechanism and use in: comprehensive molecular insect science. Elsevier BV 2005; 175-206.
  • [7] Ohba M., Aizawa K. Insect toxicity of Bacillus thuringiensis isolated from soils of Japan. J Invertebr Pathol 1986; 47: 12-20.
  • [8] Lee M.K., Walters F.S., Hart H., Palekar N., Chen J.S. The mode of action of the Bacillus thuringiensis vegatative insecticidal protein vip3A differs from that of Cry1Ab δ- endotoxin. Appl Environ Microbiol 2003; 67: 5328- 5330.
  • [9] Ohba M., Wasano, N., Mizuki, E. Bacillus thuringiensis soil populations naturally occuring in the Ryuksus a subtropic region of Japan. Microbiol Res 2000; 155: 17- 22.
  • [10] Mizuki E., Ohba M., Akao T., Yamashita S., Saitoh H., Park Y.S. Unique avtivity associated with non- insectkicidal Bacillus thuringiensis parasporal inclusions: in vitro cell- killing action on human cancer cells. J Appl Microbiol 1999; 86: 477-486.
  • [11] Guerchicoff A., Delécluse A., Rubinstein C.P. The Bacillus thuringiensis cyt genes for the hemolytic endotoxins constitude a gene family. Appl Environ Microbiol 2001; 67: 1090-1096.
  • [12] Granum P.E, Lund T. Bacillus cereus and its food poisoning toxins. FEMS Microbiol Lett 1996; 14:145-149.
  • [13] Arnesen L.P.S, Fagerlund A., Granum P.E From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev 2008; 32: 579-606.
  • [14] Mizuki E., Park Y.S., Saitoh H., Yamashita S., Akao T., Higuchi K., Ohba M. Parasporin a human leukemic cell- recognizing parasporal protein of Bacillus thuringiensis. Clin Diagn Lab Immunol 2000; 7: 624- 634.
  • [15] Katayama H., Yokota H., Akao T., Nakamura O., Ohba M., Mekada E., Mizuki E.. Parasporin-1 a novel cytotoxic protein to human cells from non-insecticidal parasporal inclusions of Bacillus thuringiensis. J Biochem 2005; 137: 17-25.
  • [16] Akiba T., Okumura S. Parasporins 1 and 2: their structure and activity. J of intervebr pathol 2016; 142: 44-49.
  • [17] Ohba M., Mizuki E., Uemori A. Parasporin, a new group of anticancer protein group from Bacillus thuringiensis. Anticancer Res 2009; 29: 427- 434.
  • [18] Comitte of Parasporin Classification and nomenclature. [http://parasporin.fitc.pref.fukuoka.jp/Accesed] (14 May 2015).
  • [19] El-hag H.A.A., Safhi M.A. Antimalignancy activity of Bacillus thuringiensis serovar dakota (H15) in vivo. J Med Sci 2011; 6: 06–16.
  • [20] Gonzalez E., Granados J.C., Short J.D., Ammons D.R., Rampersad J. Parasporins from a Caribbean island: evidence for a globally dispersed Bacillus thuringiensis strain. Curr Microbiol 2011; 62: 1643–1648.
  • [21] Poornima K., Saranya V., Abirami P., Binuramesh C., Suguna P., Selveganayagam P., Shenbagarathai R. Phenotypic and genotypic characterization of B.t.LDC- 391 strain that produce cytocidal proteins against human cancer cells. Bioinformation 2012; 8: 461–465.
  • [22] Ammons D.R., Short J.D., Bailey J., Hinojosa G., Tavarez L., Salazar M., Rampersad J.N. Anti-cancer Parasporin Toxins are Associated with Different Environments: Discovery of Two Novel Parasporin 5-like Genes. Curr Microbiol 2015; 72(2): 184-189.
  • [23] Inouye K., Okumura S., Mizuki E. Parasporin-4, a novel cancer cell-killing protein produced by Bacillus thuringiensis. Food Sci Biotechnol 2008; 17: 219–227.
  • [24] Yamashita S., Katayama H., Saitoh H., Akao T., Park Y.S., Mizuki E., Ohba M., Ito A. Typical three domain Cry proteins of Bacillus thuringiensis strain A1462 exhibit cytocidal activity on limited human cancer cells. J Biochem 2005; 138: 663- 672.
  • [25] Nagamatsu Y., Okamura S., Saitouh H., Akao Y., Mizuki E. Tree Cry toxins in two types from Bacillus thuringiensis strain M019 preferentially kill human hepatocyte cancer and uterus cervix cancer cells. Biosci Biotechnol Biochem 2010; 74 (3): 494- 498.
  • [26] Kitada S., Abe Y., Shimada H., Kusaka Y., Matsuo Y., Katayama H., Okumura S., Akao T., Mizuki E., Kuge O., Sasaguri Y., Ohba M., Ito A. Cytocidal actions of parasporin-2 an antitumor crystal toxin from Bacillus thuringiensis. J Biol Chem 2006; 281: 26350- 26360.
  • [27] Okumura S, Saitoh H., Ishikawa T., Inouye K., Mizuki E. Mode of action of parasporin-4, a cytocidal protein from Bacillus thuringiensis. BBA - Biomembranes 2011; 6: 1476–1482.
  • [28] Ekino K., Okumura S., Ishikawa T., Kitada S., Saitoh H., Akao T., Oka T., Nomura Y., Ohba M., Shin T., Mizuki E. Cloning and characterization of a unique cytotoxic protein parasporin-5 produced by Bacillus thuringiensis A1100 strain. Toxins 2014; 6(6): 1882-1895.
  • [29] Katayama H., Kusaka Y., Yokota H., Akao T., Kojima M., Nakamura O., Mekada E., Mizuki E. Parasporin-1 a novel cytotoxic protein from Bacillus thuringiensis, induces Ca++ influx and a sustained elevation of the cytoplasmic Ca++ concentration in toxin- sensitive cells. J Biol Chem 2007; 282: 7742- 7752.
  • [30] Katayama H., Kusaka Y., Mizuki E. Parasproin-1 receptor and use thereof. European Patent 2273266, filed 30 March 2009, issued 12 January 2011.
  • [31] Ito A., Sasaguri Y., Kitada S., Kusaka Y., Kuwano K., Masutomi K., Mizuki E., Akao T., Ohba M. A Bacillus thuringiensis crystal protein with selective cytocidal action to human cells. J Biol Chem 2004; 279: 21282- 21286.
  • [32] Brown K.L., Whiteley H.R. Molecular characterization of two novel crystal protein genes from Bacillus thuringiensis subsp. thompsoni. J Bacteriol 1992; 174:549- 557.
  • [33] Brasseur K., Auger P., Asselin E., Parent S., Côté J.C, Sirois M. Parasporin-2 from a New Bacillus thuringiensis 4R2 Strain Induces Caspases Activation and Apoptosis in Human Cancer Cells. PLoS One 2015; 10(8): e0135106.
  • [34] Kitada S., Abe Y., Maeda T., Shimada H. Parasporin-2 requires GPI-anchored proteins for the efficient cytocidal action to human hepatoma cells. Toxicol 2010; 264: 80–88.
  • [35] Shimada H., Kitada S. Mega assemblages of oligomeric aerolysin-like toxins stabilized by toxin-associating membrane proteins. J. Biochem 2011: 149; 103–115.
  • [36] Okumura S., Akao T., Higuchi K., Saitoh H., Mizuki E., Ohba M., Inouye K. Bacillus thuringiensis serovar shandongiensis strain 89-T-34-22 produces multiple cytotoxic proteins with similar molecular masses against human cancer cells. Lett Appl Microbiol 2004; 39:89-92
  • [37] Okassov A. Parasporins as a new anticancer agents: a review. J Buon 2015; 20(1): 5-16.
  • [38] Krishnan V. Investigation of parasporins, the cytotoxic proteins from the bacterium Bacillus thuringiensis. Thesis: Department of Biochemistry, School of Life Sciences, University of Sussex, Sussex, 2013.
  • [39] Poornima K., Selvanayagam P., Shenbagarathai R. Identification of native Bacillus thuringiensis strain from South India having specific cytocidal activity against cancer cells. J Appl Microbiol 2010; 109:348-354.

DOĞAL BİR ANTİ-KANSER PROTEİN OLAN PARASPORİNLER

Year 2017, Volume: 6 Issue: 2, 103 - 113, 31.08.2017
https://doi.org/10.18036/aubtdc.307261

Abstract

Bacillus thuringiensis doğada
geniş yayılım gösteren, gram pozitif ve spor oluşturan bir bakteridir.
Sporulasyon evresinde, sporlarla birlikte birçok böcek çeşidine karşı
insektisit etki gösteren kristal proteinleri de üretir. İnsektisidal kristal
proteinler Cry ve Cyt proteinler olup; aktivitelerini duyarlı böceklerin orta
barsak membranlarında bulunan reseptöre bağlanarak gösterirler. Bu reseptör
bağlantılı özgül aktivitelerinden dolayı zirai mücadelede önemlidirler. Daha
sonraları, insektisit olanlara göre doğada daha geniş dağılım gösteren
insektisit olmayan kristal proteinler üzerindeki çalışmalar yeni bir protein
sınıfı olan parasporinleri ortaya çıkarmıştır. Parasporinler ayrıcalıklı olarak
kanser hücrelerine karşı sitotoksik etki gösterirken normal hücrelere etki
etmez ve hemolitik etki göstermez. Şimdiye kadar altı parasporin ailesi
tanımlanmış olup toplamda 19 alt grup bilinmektedir. Parasporinler de
insektisidal kristal proteinlere benzer şekilde reseptör bağlantılı etki
gösterdiğinden kanser tedavisiyle ilgili çalışmalarda yeni bir araştırma
alanının ortaya çıkmasını sağlamıştır. Bu derlemede; parasporinlerin keşfi,
özellikleri, çeşitleri ve etki mekanizmaları hakkında yapılmış olan çalışmalar özetle
anlatılmıştır.

References

  • KAYNAKLAR [1] Ishiwata S. On a kind of severe flacherie (sotto disease). Dainihon Sanshi Kaiho 1901; 114: 1-5.
  • [2] Ben- Dov E., Zaritsky A., Dahan E., Barak Z., Sınal R., Manasherob R., Khamraev A., Troitskaya E., Dubitsky A., Berezina N., Margalith Y. Extended screening by PCR for seven cry-group genes from field collected strains of Bacillus thuringienis. Appl Environ Microbiol 1997; 63: 4883- 4890.
  • [3] Bernhard K., Jarrett P., Meadows M., Butt J., Ellis D.J., Roberts G.M., Pauli S., Rodgers P., Burges H.D. Natural Isolates Of Bacillus Thuringiensis: Worldwide Distribution, Characterization, And Activity Against Insect Pests. J Invertebr Pathol 1997; 70 (1): 59-68.
  • [4] Ichimatsu T., Mizuki E., Nishimura K., Akao T., Saitoh H., Higuchi K., Ohba M. Occurrence of Bacillus thuringiensis in fresh water of Japan. Curr Microbiol 2000; 40: 212- 217.
  • [5] Logan N. Bacillus anthracis, Bacillus cereus, and other aerobic endospore-forming bacteria. Topley & Wilson’s Microbiology & Microbial Infections (Tenth Edition). Bacteriology. Borriello SP, Murray PR and Funke G (eds.). London: Hodder Arnold; 922-952, 2005.
  • [6] Bravo A., Gill S.S., Soberon M. Bacillus thuringiensis mechanism and use in: comprehensive molecular insect science. Elsevier BV 2005; 175-206.
  • [7] Ohba M., Aizawa K. Insect toxicity of Bacillus thuringiensis isolated from soils of Japan. J Invertebr Pathol 1986; 47: 12-20.
  • [8] Lee M.K., Walters F.S., Hart H., Palekar N., Chen J.S. The mode of action of the Bacillus thuringiensis vegatative insecticidal protein vip3A differs from that of Cry1Ab δ- endotoxin. Appl Environ Microbiol 2003; 67: 5328- 5330.
  • [9] Ohba M., Wasano, N., Mizuki, E. Bacillus thuringiensis soil populations naturally occuring in the Ryuksus a subtropic region of Japan. Microbiol Res 2000; 155: 17- 22.
  • [10] Mizuki E., Ohba M., Akao T., Yamashita S., Saitoh H., Park Y.S. Unique avtivity associated with non- insectkicidal Bacillus thuringiensis parasporal inclusions: in vitro cell- killing action on human cancer cells. J Appl Microbiol 1999; 86: 477-486.
  • [11] Guerchicoff A., Delécluse A., Rubinstein C.P. The Bacillus thuringiensis cyt genes for the hemolytic endotoxins constitude a gene family. Appl Environ Microbiol 2001; 67: 1090-1096.
  • [12] Granum P.E, Lund T. Bacillus cereus and its food poisoning toxins. FEMS Microbiol Lett 1996; 14:145-149.
  • [13] Arnesen L.P.S, Fagerlund A., Granum P.E From soil to gut: Bacillus cereus and its food poisoning toxins. FEMS Microbiol Rev 2008; 32: 579-606.
  • [14] Mizuki E., Park Y.S., Saitoh H., Yamashita S., Akao T., Higuchi K., Ohba M. Parasporin a human leukemic cell- recognizing parasporal protein of Bacillus thuringiensis. Clin Diagn Lab Immunol 2000; 7: 624- 634.
  • [15] Katayama H., Yokota H., Akao T., Nakamura O., Ohba M., Mekada E., Mizuki E.. Parasporin-1 a novel cytotoxic protein to human cells from non-insecticidal parasporal inclusions of Bacillus thuringiensis. J Biochem 2005; 137: 17-25.
  • [16] Akiba T., Okumura S. Parasporins 1 and 2: their structure and activity. J of intervebr pathol 2016; 142: 44-49.
  • [17] Ohba M., Mizuki E., Uemori A. Parasporin, a new group of anticancer protein group from Bacillus thuringiensis. Anticancer Res 2009; 29: 427- 434.
  • [18] Comitte of Parasporin Classification and nomenclature. [http://parasporin.fitc.pref.fukuoka.jp/Accesed] (14 May 2015).
  • [19] El-hag H.A.A., Safhi M.A. Antimalignancy activity of Bacillus thuringiensis serovar dakota (H15) in vivo. J Med Sci 2011; 6: 06–16.
  • [20] Gonzalez E., Granados J.C., Short J.D., Ammons D.R., Rampersad J. Parasporins from a Caribbean island: evidence for a globally dispersed Bacillus thuringiensis strain. Curr Microbiol 2011; 62: 1643–1648.
  • [21] Poornima K., Saranya V., Abirami P., Binuramesh C., Suguna P., Selveganayagam P., Shenbagarathai R. Phenotypic and genotypic characterization of B.t.LDC- 391 strain that produce cytocidal proteins against human cancer cells. Bioinformation 2012; 8: 461–465.
  • [22] Ammons D.R., Short J.D., Bailey J., Hinojosa G., Tavarez L., Salazar M., Rampersad J.N. Anti-cancer Parasporin Toxins are Associated with Different Environments: Discovery of Two Novel Parasporin 5-like Genes. Curr Microbiol 2015; 72(2): 184-189.
  • [23] Inouye K., Okumura S., Mizuki E. Parasporin-4, a novel cancer cell-killing protein produced by Bacillus thuringiensis. Food Sci Biotechnol 2008; 17: 219–227.
  • [24] Yamashita S., Katayama H., Saitoh H., Akao T., Park Y.S., Mizuki E., Ohba M., Ito A. Typical three domain Cry proteins of Bacillus thuringiensis strain A1462 exhibit cytocidal activity on limited human cancer cells. J Biochem 2005; 138: 663- 672.
  • [25] Nagamatsu Y., Okamura S., Saitouh H., Akao Y., Mizuki E. Tree Cry toxins in two types from Bacillus thuringiensis strain M019 preferentially kill human hepatocyte cancer and uterus cervix cancer cells. Biosci Biotechnol Biochem 2010; 74 (3): 494- 498.
  • [26] Kitada S., Abe Y., Shimada H., Kusaka Y., Matsuo Y., Katayama H., Okumura S., Akao T., Mizuki E., Kuge O., Sasaguri Y., Ohba M., Ito A. Cytocidal actions of parasporin-2 an antitumor crystal toxin from Bacillus thuringiensis. J Biol Chem 2006; 281: 26350- 26360.
  • [27] Okumura S, Saitoh H., Ishikawa T., Inouye K., Mizuki E. Mode of action of parasporin-4, a cytocidal protein from Bacillus thuringiensis. BBA - Biomembranes 2011; 6: 1476–1482.
  • [28] Ekino K., Okumura S., Ishikawa T., Kitada S., Saitoh H., Akao T., Oka T., Nomura Y., Ohba M., Shin T., Mizuki E. Cloning and characterization of a unique cytotoxic protein parasporin-5 produced by Bacillus thuringiensis A1100 strain. Toxins 2014; 6(6): 1882-1895.
  • [29] Katayama H., Kusaka Y., Yokota H., Akao T., Kojima M., Nakamura O., Mekada E., Mizuki E. Parasporin-1 a novel cytotoxic protein from Bacillus thuringiensis, induces Ca++ influx and a sustained elevation of the cytoplasmic Ca++ concentration in toxin- sensitive cells. J Biol Chem 2007; 282: 7742- 7752.
  • [30] Katayama H., Kusaka Y., Mizuki E. Parasproin-1 receptor and use thereof. European Patent 2273266, filed 30 March 2009, issued 12 January 2011.
  • [31] Ito A., Sasaguri Y., Kitada S., Kusaka Y., Kuwano K., Masutomi K., Mizuki E., Akao T., Ohba M. A Bacillus thuringiensis crystal protein with selective cytocidal action to human cells. J Biol Chem 2004; 279: 21282- 21286.
  • [32] Brown K.L., Whiteley H.R. Molecular characterization of two novel crystal protein genes from Bacillus thuringiensis subsp. thompsoni. J Bacteriol 1992; 174:549- 557.
  • [33] Brasseur K., Auger P., Asselin E., Parent S., Côté J.C, Sirois M. Parasporin-2 from a New Bacillus thuringiensis 4R2 Strain Induces Caspases Activation and Apoptosis in Human Cancer Cells. PLoS One 2015; 10(8): e0135106.
  • [34] Kitada S., Abe Y., Maeda T., Shimada H. Parasporin-2 requires GPI-anchored proteins for the efficient cytocidal action to human hepatoma cells. Toxicol 2010; 264: 80–88.
  • [35] Shimada H., Kitada S. Mega assemblages of oligomeric aerolysin-like toxins stabilized by toxin-associating membrane proteins. J. Biochem 2011: 149; 103–115.
  • [36] Okumura S., Akao T., Higuchi K., Saitoh H., Mizuki E., Ohba M., Inouye K. Bacillus thuringiensis serovar shandongiensis strain 89-T-34-22 produces multiple cytotoxic proteins with similar molecular masses against human cancer cells. Lett Appl Microbiol 2004; 39:89-92
  • [37] Okassov A. Parasporins as a new anticancer agents: a review. J Buon 2015; 20(1): 5-16.
  • [38] Krishnan V. Investigation of parasporins, the cytotoxic proteins from the bacterium Bacillus thuringiensis. Thesis: Department of Biochemistry, School of Life Sciences, University of Sussex, Sussex, 2013.
  • [39] Poornima K., Selvanayagam P., Shenbagarathai R. Identification of native Bacillus thuringiensis strain from South India having specific cytocidal activity against cancer cells. J Appl Microbiol 2010; 109:348-354.
There are 39 citations in total.

Details

Journal Section Review
Authors

Müjgan Kesik Oktay

Hatice Güneş

Publication Date August 31, 2017
Published in Issue Year 2017 Volume: 6 Issue: 2

Cite

APA Kesik Oktay, M., & Güneş, H. (2017). PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology, 6(2), 103-113. https://doi.org/10.18036/aubtdc.307261
AMA Kesik Oktay M, Güneş H. PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology. August 2017;6(2):103-113. doi:10.18036/aubtdc.307261
Chicago Kesik Oktay, Müjgan, and Hatice Güneş. “PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN”. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology 6, no. 2 (August 2017): 103-13. https://doi.org/10.18036/aubtdc.307261.
EndNote Kesik Oktay M, Güneş H (August 1, 2017) PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology 6 2 103–113.
IEEE M. Kesik Oktay and H. Güneş, “PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN”, Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology, vol. 6, no. 2, pp. 103–113, 2017, doi: 10.18036/aubtdc.307261.
ISNAD Kesik Oktay, Müjgan - Güneş, Hatice. “PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN”. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology 6/2 (August 2017), 103-113. https://doi.org/10.18036/aubtdc.307261.
JAMA Kesik Oktay M, Güneş H. PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology. 2017;6:103–113.
MLA Kesik Oktay, Müjgan and Hatice Güneş. “PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN”. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology, vol. 6, no. 2, 2017, pp. 103-1, doi:10.18036/aubtdc.307261.
Vancouver Kesik Oktay M, Güneş H. PARASPORINS AS A NATURAL ANTI-CANCER PROTEIN. Anadolu University Journal of Science and Technology C - Life Sciences and Biotechnology. 2017;6(2):103-1.