Araştırma Makalesi
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Optimization of recombinant novel esterase expression from extremophiles

Yıl 2018, Cilt: 12 Sayı: 3, 33 - 36, 26.12.2018

Havva Esra Tütüncü Nurgul Çelik Balci Melek Tuter Nevin Gul Karaguler

Öz

Esterases, which
are a sub-group of lipolytic enzymes, are important biocatalysts for many
industrial applications. In this study, optimization for the recombinant
expression of a novel esterase, which was previously obtained by metagenomic
approach, was studied. To optimize the expression, 0.1, 0.5 and 1 mM isopropyl
β-D-1 thiogalactopyranoside (IPTG) concentrations were tried. In addition,
induction at 25ºC for 16 hours, 30ºC for 6 hours and 37ºC for 3 hours were
tried. According to the results, induction at 30°C for 6 hours by 0.1 mM IPTG yielded
high amount of protein with maximum catalytic activity. After the gene was
successfully expressed, purification studies were conducted. The protein was
purified using His-tag method. Native and SDS-PAGE analysis showed that protein
which is present as a monomer was successfully purified. 

Anahtar Kelimeler

Protein expression esterase

Kaynakça

  • [1] Villeneuve P, Muderhwa, JM, Graille, J & Haas, MJ. 2000. Customizing lipases for biocatalysis: a survey of chemical, physical and molecular biological approaches. Journal of molecular catalysis B: enzymatic, 9(4-6), 113-148.
  • [2] Arpigny, JL & Jaeger, KE. 1999. Bacterial lipolytic enzymes: classification and properties. Biochemical journal, 343(Pt 1), 177.
  • [3] Handelsman, J, Rondon, MR, Brady, SF, Clardy, J, & Goodman, RM. 1998. Molecular biological access to the chemistry of unknown soil microbes: a new frontier for natural products. Chemistry & Biology, 5(10), R245–R249.
  • [4] Oren, A. 2008. Microbial life at high salt concentrations: phylogenetic and metabolic diversity. Saline Systems, 4(1), 2.
  • [5] Bahreini, E, Aghaiypour, K, Abbasalipourkabir, R, Goodarzi, MT, Saidijam, M & Safavieh, SS. 2014. An optimized protocol for overproduction of recombinant protein expression in Escherichia coli. Preparative Biochemistry and Biotechnology,44(5), 510-528.
  • [6] Rosano, GL, & Ceccarelli, EA. 2014. Recombinant protein expression in Escherichia coli: advances and challenges. Frontiers in microbiology, 5, 172.
  • [7] Miroux, B, & Walker, JE. 1996. Over-production of Proteins in Escherichia coli: Mutant Hosts that Allow Synthesis of some Membrane Proteins and Globular Proteins at High Levels. Journal of Molecular Biology, 260(3), 289–298.
  • [8] Graumann, K, & Premstaller, A. 2006. Manufacturing of recombinant therapeutic proteins in microbial systems. Biotechnol. J. 1, 164–186.
  • [9] Sørensen, HP & Mortensen, KK. 2005. Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli. Microbial cell factories, 4(1), 1.
  • [10] Tutuncu HE. 2017. Strategies for Isolation of Novel Enzymes Using Metagenomics Approach, PhD Thesis, YOK Thesis number: 465429.
  • [11] Bradford, MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2), 248-254.
  • [12] Winkler, UK. & Stuckmann, M. 1979. Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens. Journal of Bacteriology, 138(3), 663–670.
  • [13] Pereira, MR, Maester, TC, Mercaldi, GF, de Macedo Lemos, EG, Hyvönen, M. & Balan, A. 2017. From a metagenomic source to a high-resolution structure of a novel alkaline esterase. Applied microbiology and biotechnology, 101(12), 4935-4949.
  • [14] Zhang, Y, Hao, J, Zhang, YQ, Chen, XL, Xie, BB, Shi, M, ... & Li, PY. 2017. Identification and characterization of a novel salt-tolerant esterase from the deep-sea sediment of the South China Sea. Frontiers in microbiology, 8, 441.
  • [15] Maester, TC, Pereira, MR, Sierra, EM, Balan, A, & de Macedo Lemos, EG. 2016. Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications. Applied microbiology and biotechnology, 100(13), 5815-5827.
  • [16] Bornscheuer, UT. 2002. Microbial carboxyl esterases: Classification, properties and application in biocatalysis. FEMS Microbiology Reviews, 26(1), 73–81.

Yıl 2018, Cilt: 12 Sayı: 3, 33 - 36, 26.12.2018

Havva Esra Tütüncü Nurgul Çelik Balci Melek Tuter Nevin Gul Karaguler

Öz

Kaynakça

  • [1] Villeneuve P, Muderhwa, JM, Graille, J & Haas, MJ. 2000. Customizing lipases for biocatalysis: a survey of chemical, physical and molecular biological approaches. Journal of molecular catalysis B: enzymatic, 9(4-6), 113-148.
  • [2] Arpigny, JL & Jaeger, KE. 1999. Bacterial lipolytic enzymes: classification and properties. Biochemical journal, 343(Pt 1), 177.
  • [3] Handelsman, J, Rondon, MR, Brady, SF, Clardy, J, & Goodman, RM. 1998. Molecular biological access to the chemistry of unknown soil microbes: a new frontier for natural products. Chemistry & Biology, 5(10), R245–R249.
  • [4] Oren, A. 2008. Microbial life at high salt concentrations: phylogenetic and metabolic diversity. Saline Systems, 4(1), 2.
  • [5] Bahreini, E, Aghaiypour, K, Abbasalipourkabir, R, Goodarzi, MT, Saidijam, M & Safavieh, SS. 2014. An optimized protocol for overproduction of recombinant protein expression in Escherichia coli. Preparative Biochemistry and Biotechnology,44(5), 510-528.
  • [6] Rosano, GL, & Ceccarelli, EA. 2014. Recombinant protein expression in Escherichia coli: advances and challenges. Frontiers in microbiology, 5, 172.
  • [7] Miroux, B, & Walker, JE. 1996. Over-production of Proteins in Escherichia coli: Mutant Hosts that Allow Synthesis of some Membrane Proteins and Globular Proteins at High Levels. Journal of Molecular Biology, 260(3), 289–298.
  • [8] Graumann, K, & Premstaller, A. 2006. Manufacturing of recombinant therapeutic proteins in microbial systems. Biotechnol. J. 1, 164–186.
  • [9] Sørensen, HP & Mortensen, KK. 2005. Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli. Microbial cell factories, 4(1), 1.
  • [10] Tutuncu HE. 2017. Strategies for Isolation of Novel Enzymes Using Metagenomics Approach, PhD Thesis, YOK Thesis number: 465429.
  • [11] Bradford, MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2), 248-254.
  • [12] Winkler, UK. & Stuckmann, M. 1979. Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens. Journal of Bacteriology, 138(3), 663–670.
  • [13] Pereira, MR, Maester, TC, Mercaldi, GF, de Macedo Lemos, EG, Hyvönen, M. & Balan, A. 2017. From a metagenomic source to a high-resolution structure of a novel alkaline esterase. Applied microbiology and biotechnology, 101(12), 4935-4949.
  • [14] Zhang, Y, Hao, J, Zhang, YQ, Chen, XL, Xie, BB, Shi, M, ... & Li, PY. 2017. Identification and characterization of a novel salt-tolerant esterase from the deep-sea sediment of the South China Sea. Frontiers in microbiology, 8, 441.
  • [15] Maester, TC, Pereira, MR, Sierra, EM, Balan, A, & de Macedo Lemos, EG. 2016. Characterization of EST3: a metagenome-derived esterase with suitable properties for biotechnological applications. Applied microbiology and biotechnology, 100(13), 5815-5827.
  • [16] Bornscheuer, UT. 2002. Microbial carboxyl esterases: Classification, properties and application in biocatalysis. FEMS Microbiology Reviews, 26(1), 73–81.
Toplam 16 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Bölüm Araştırma Makalesi
Yazarlar

Havva Esra Tütüncü

Nurgul Çelik Balci

Melek Tuter

Nevin Gul Karaguler Bu kişi benim

Yayımlanma Tarihi 26 Aralık 2018
Yayımlandığı Sayı Yıl 2018 Cilt: 12 Sayı: 3

Kaynak Göster

APA Tütüncü, H. E., Çelik Balci, N., Tuter, M., Gul Karaguler, N. (2018). Optimization of recombinant novel esterase expression from extremophiles. Journal of Applied Biological Sciences, 12(3), 33-36.
AMA Tütüncü HE, Çelik Balci N, Tuter M, Gul Karaguler N. Optimization of recombinant novel esterase expression from extremophiles. J.appl.biol.sci. Aralık 2018;12(3):33-36.
Chicago Tütüncü, Havva Esra, Nurgul Çelik Balci, Melek Tuter, ve Nevin Gul Karaguler. “Optimization of Recombinant Novel Esterase Expression from Extremophiles”. Journal of Applied Biological Sciences 12, sy. 3 (Aralık 2018): 33-36.
EndNote Tütüncü HE, Çelik Balci N, Tuter M, Gul Karaguler N (01 Aralık 2018) Optimization of recombinant novel esterase expression from extremophiles. Journal of Applied Biological Sciences 12 3 33–36.
IEEE H. E. Tütüncü, N. Çelik Balci, M. Tuter, ve N. Gul Karaguler, “Optimization of recombinant novel esterase expression from extremophiles”, J.appl.biol.sci., c. 12, sy. 3, ss. 33–36, 2018.
ISNAD Tütüncü, Havva Esra vd. “Optimization of Recombinant Novel Esterase Expression from Extremophiles”. Journal of Applied Biological Sciences 12/3 (Aralık 2018), 33-36.
JAMA Tütüncü HE, Çelik Balci N, Tuter M, Gul Karaguler N. Optimization of recombinant novel esterase expression from extremophiles. J.appl.biol.sci. 2018;12:33–36.
MLA Tütüncü, Havva Esra vd. “Optimization of Recombinant Novel Esterase Expression from Extremophiles”. Journal of Applied Biological Sciences, c. 12, sy. 3, 2018, ss. 33-36.
Vancouver Tütüncü HE, Çelik Balci N, Tuter M, Gul Karaguler N. Optimization of recombinant novel esterase expression from extremophiles. J.appl.biol.sci. 2018;12(3):33-6.
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