Araştırma Makalesi
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GFP Raportör Proteini Kullanarak E. coli'de Rekombinant Protein Üretiminde Peynir Altı Suyu Tozunun İndükleyici Rolünün Doğrulanması

Yıl 2026, Cilt: 15 Sayı: 1 , 58 - 66 , 30.03.2026
https://doi.org/10.46810/tdfd.1752790
https://izlik.org/JA57GH58SF

Öz

Peynir altı suyu, süt endüstrisinden elde edilen değerli bir yan üründür. Son zamanlarda yapılan çalışmalar, peynir altı suyunun rekombinant protein üretiminde hem indükleyici hem de karbon kaynağı olarak kullanımına odaklanmaktadır. Bu çalışmanın amacı, Escherichia coli'den üretilen rekombinant yeşil floresan proteininin indükleyicisi olarak tatlı peynir altı suyu tozunda bulunan laktozun etkinliğini ve minimum konsantrasyonunu değerlendirmektir. Peynir altı suyunun potansiyeli optik yoğunluk, hücre biyokütlesi, elektroforez analizi, floresans, protein saflığı temelinde değerlendirilmiş ve sonuçlar IPTG'den elde edilenlerle karşılaştırılmıştır. En yüksek floresans değerleri 20 mM laktoz eşdeğerinde deproteinize peynir altı suyu tozu ile elde edilmiştir. 2 mM laktoz içeren deproteinize peynir altı suyu tozu kullanımı, 0,5 mM IPTG'ye benzer miktarlarda rekombinant GFP üretmek için yeterli olmuştur. Peynir altı suyu tozunun bir indükleyici olarak doğrulanması, yüksek miktarda IPTG kullanımına bağlı toksisite ve ekonomik sorunları ortadan kaldıracaktır. Ayrıca, bu strateji peynir altı suyunun neden olduğu çevre kirliliği sorunları için alternatif bir çözüm sunmaktadır.

Kaynakça

  • Boumaiza M, Colarusso A, Parrilli E, Garcia-Fruitós E, Casillo A, Arís A, Corsaro MM, Picone D, Leone S, Tutino ML. Getting value from the waste: Recombinant production of a sweet protein by Lactococcus lactis grown on cheese whey. Microb Cell Fact. 2018;17(1):1–10.
  • Mehri D, Perendeci NA, Goksungur Y. Utilization of whey for red pigment production by Monascus purpureus in submerged fermentation. Fermentation. 2021;7(2).
  • Królczyk JB, Dawidziuk T, Janiszewska-Turak E, Sołowiej B. Use of Whey and Whey Preparations in the Food Industry – a Review. Polish J Food Nutr Sci. 2016 Sep 30;66(3):157–65.
  • Chourasia R, Phukon LC, Abedin MM, Padhi S, Singh SP, Rai AK. Whey valorization by microbial and enzymatic bioprocesses for the production of nutraceuticals and value-added products. Bioresour Technol Reports. 2022 Sep 1;19:101144.
  • Yadav JSS, Yan S, Pilli S, Kumar L, Tyagi RD, Surampalli RY. Cheese whey: A potential resource to transform into bioprotein, functional/nutritional proteins and bioactive peptides. Biotechnol Adv. 2015 Nov 1;33(6):756–74.
  • Hausjell J, Miltner M, Herzig C, Limbeck A, Saracevic Z, Saracevic E, Weissensteiner J, Molitor C, Halbwirth H, Spadiut O. Valorisation of cheese whey as substrate and inducer for recombinant protein production in E. coli HMS174(DE3). Bioresour Technol Reports. 2019;8(October):100340.
  • Al-Hawash AB, Zhang X, Ma F. Strategies of codon optimization for high-level heterologous protein expression in microbial expression systems. Gene Reports. 2017 Dec 1;9:46–53.
  • Donovan RS, Robinson CW, Glick BR. Review: Optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter. J Ind Microbiol. 1996;16:145–54.
  • Dvorak P, Chrast L, Nikel PI, Fedr R, Soucek K, Sedlackova M, Chaloupkova R, Lorenzo V, Prokop Z, Damborsky J. Exacerbation of substrate toxicity by IPTG in Escherichia coli BL21(DE3) carrying a synthetic metabolic pathway. Microb Cell Fact. 2015 Dec 21;14(1):201.
  • Menzella HG, Ceccarelli EA, Gramajo HC. Novel Escherichia coli strain allows efficient recombinant protein production using lactose as inducer. Biotechnol Bioeng. 2003 Jun 30;82(7):809–17.
  • Lavari L, Páez R, Cuatrin A, Reinheimer J, Vinderola G. Use of cheese whey for biomass production and spray drying of probiotic lactobacilli. J Dairy Res. 2014;81(3):267–74.
  • De León-Rodríguez A, Rivera-Pastrana D, Medina-Rivero E, Flores-Flores JL, Estrada-Baltazar A, Ordóñez-Acevedo LG, de la Rosa APB. Production of penicillin acylase by a recombinant Escherichia coli using cheese whey as substrate and inducer. Biomol Eng. 2006;23(6):299–305.
  • Mobayed FH, Nunes JC, Gennari A, de Andrade BC, Ferreira MLV, Pauli P, Renard G, Chies JM, Volpato G, Volken de Souza CF. Effect of by-products from the dairy industry as alternative inducers of recombinant β-galactosidase expression. Biotechnol Lett. 2021;43(3):589–99.
  • Viitanen MI, Vasala A, Neubauer P, Alatossava T. Cheese whey-induced high-cell-density production of recombinant proteins in Escherichia coli. Microb Cell Fact. 2003;2(2):1–10.
  • Ding J, You S, Ba W, Zhang H, Chang H, Qi W, Su R, He Z. Bifunctional utilization of whey powder as a substrate and inducer for β-farnesene production in an engineered Escherichia coli. Bioresour Technol. 2021 Dec 1;341:125739.
  • Greicius A, Baliutavicius T, Lastauskiene E, Gudiukaite R. Application of Milk Permeate as an Inducer for the Production of Microbial Recombinant Lipolytic Enzymes. Ferment 2023, Vol 9, Page 27. 2022 Dec 28;9(1):27.
  • Albano CR, Randers-Eichhorn L, Bentley WE, Rao G. Green Fluorescent Protein as a Real Time Quantitative Reporter of Heterologous Protein Production. 1998;
  • Drew DE, Von Heijne G, Nordlund P, De Gier JWL. Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli. FEBS Lett. 2001 Oct 26;507(2):220–4.
  • de Divitiis M, Ami D, Pessina A, Palmioli A, Sciandrone B, Airoldi C, Regonesi ME, Brambilla L, Lotti M, Natalello A, Brocca S, Mangiagalli M. Cheese-whey permeate improves the fitness of Escherichia coli cells during recombinant protein production. Biotechnol Biofuels Bioprod. 2023;16(1):1–19.

Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein

Yıl 2026, Cilt: 15 Sayı: 1 , 58 - 66 , 30.03.2026
https://doi.org/10.46810/tdfd.1752790
https://izlik.org/JA57GH58SF

Öz

Whey is a valuable by-product obtained from the dairy industry. Recently studies focus on utilization of whey as both inducer and carbon source in recombinant protein production. The aim of the present study was to evaluate the efficiency and minimum concentration of lactose present in sweet whey powder as inducer of recombinant green fluorescent protein produced from Escherichia coli. The potential of whey was evaluated based on optical density, cell biomass, electrophoresis analysis, fluorescence, protein purity and results were compared with those obtained from IPTG. The highest fluorescence values were obtained with 20 mM lactose equivalent of deproteinized whey powder. Use of 2 mM lactose containing deproteinized whey powder was sufficient to produce recombinant GFP at comparable amounts to 0.5 mM IPTG. Validation of whey powder as an inducer will eliminate the toxicity and economic problems associated with high amount of IPTG usage. Furthermore, this strategy provides an alternative solution for the environmental pollution problems caused by whey.

Kaynakça

  • Boumaiza M, Colarusso A, Parrilli E, Garcia-Fruitós E, Casillo A, Arís A, Corsaro MM, Picone D, Leone S, Tutino ML. Getting value from the waste: Recombinant production of a sweet protein by Lactococcus lactis grown on cheese whey. Microb Cell Fact. 2018;17(1):1–10.
  • Mehri D, Perendeci NA, Goksungur Y. Utilization of whey for red pigment production by Monascus purpureus in submerged fermentation. Fermentation. 2021;7(2).
  • Królczyk JB, Dawidziuk T, Janiszewska-Turak E, Sołowiej B. Use of Whey and Whey Preparations in the Food Industry – a Review. Polish J Food Nutr Sci. 2016 Sep 30;66(3):157–65.
  • Chourasia R, Phukon LC, Abedin MM, Padhi S, Singh SP, Rai AK. Whey valorization by microbial and enzymatic bioprocesses for the production of nutraceuticals and value-added products. Bioresour Technol Reports. 2022 Sep 1;19:101144.
  • Yadav JSS, Yan S, Pilli S, Kumar L, Tyagi RD, Surampalli RY. Cheese whey: A potential resource to transform into bioprotein, functional/nutritional proteins and bioactive peptides. Biotechnol Adv. 2015 Nov 1;33(6):756–74.
  • Hausjell J, Miltner M, Herzig C, Limbeck A, Saracevic Z, Saracevic E, Weissensteiner J, Molitor C, Halbwirth H, Spadiut O. Valorisation of cheese whey as substrate and inducer for recombinant protein production in E. coli HMS174(DE3). Bioresour Technol Reports. 2019;8(October):100340.
  • Al-Hawash AB, Zhang X, Ma F. Strategies of codon optimization for high-level heterologous protein expression in microbial expression systems. Gene Reports. 2017 Dec 1;9:46–53.
  • Donovan RS, Robinson CW, Glick BR. Review: Optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter. J Ind Microbiol. 1996;16:145–54.
  • Dvorak P, Chrast L, Nikel PI, Fedr R, Soucek K, Sedlackova M, Chaloupkova R, Lorenzo V, Prokop Z, Damborsky J. Exacerbation of substrate toxicity by IPTG in Escherichia coli BL21(DE3) carrying a synthetic metabolic pathway. Microb Cell Fact. 2015 Dec 21;14(1):201.
  • Menzella HG, Ceccarelli EA, Gramajo HC. Novel Escherichia coli strain allows efficient recombinant protein production using lactose as inducer. Biotechnol Bioeng. 2003 Jun 30;82(7):809–17.
  • Lavari L, Páez R, Cuatrin A, Reinheimer J, Vinderola G. Use of cheese whey for biomass production and spray drying of probiotic lactobacilli. J Dairy Res. 2014;81(3):267–74.
  • De León-Rodríguez A, Rivera-Pastrana D, Medina-Rivero E, Flores-Flores JL, Estrada-Baltazar A, Ordóñez-Acevedo LG, de la Rosa APB. Production of penicillin acylase by a recombinant Escherichia coli using cheese whey as substrate and inducer. Biomol Eng. 2006;23(6):299–305.
  • Mobayed FH, Nunes JC, Gennari A, de Andrade BC, Ferreira MLV, Pauli P, Renard G, Chies JM, Volpato G, Volken de Souza CF. Effect of by-products from the dairy industry as alternative inducers of recombinant β-galactosidase expression. Biotechnol Lett. 2021;43(3):589–99.
  • Viitanen MI, Vasala A, Neubauer P, Alatossava T. Cheese whey-induced high-cell-density production of recombinant proteins in Escherichia coli. Microb Cell Fact. 2003;2(2):1–10.
  • Ding J, You S, Ba W, Zhang H, Chang H, Qi W, Su R, He Z. Bifunctional utilization of whey powder as a substrate and inducer for β-farnesene production in an engineered Escherichia coli. Bioresour Technol. 2021 Dec 1;341:125739.
  • Greicius A, Baliutavicius T, Lastauskiene E, Gudiukaite R. Application of Milk Permeate as an Inducer for the Production of Microbial Recombinant Lipolytic Enzymes. Ferment 2023, Vol 9, Page 27. 2022 Dec 28;9(1):27.
  • Albano CR, Randers-Eichhorn L, Bentley WE, Rao G. Green Fluorescent Protein as a Real Time Quantitative Reporter of Heterologous Protein Production. 1998;
  • Drew DE, Von Heijne G, Nordlund P, De Gier JWL. Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli. FEBS Lett. 2001 Oct 26;507(2):220–4.
  • de Divitiis M, Ami D, Pessina A, Palmioli A, Sciandrone B, Airoldi C, Regonesi ME, Brambilla L, Lotti M, Natalello A, Brocca S, Mangiagalli M. Cheese-whey permeate improves the fitness of Escherichia coli cells during recombinant protein production. Biotechnol Biofuels Bioprod. 2023;16(1):1–19.
Toplam 19 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Biyomühendislik (Diğer)
Bölüm Araştırma Makalesi
Yazarlar

Seyhan İçier 0000-0002-0627-733X

Fatma Fındık 0000-0001-9165-3653

Yekta Göksungur 0000-0002-5166-6561

Burcu Kaplan Türköz 0000-0003-3040-3321

Gönderilme Tarihi 31 Temmuz 2025
Kabul Tarihi 17 Aralık 2025
Yayımlanma Tarihi 30 Mart 2026
DOI https://doi.org/10.46810/tdfd.1752790
IZ https://izlik.org/JA57GH58SF
Yayımlandığı Sayı Yıl 2026 Cilt: 15 Sayı: 1

Kaynak Göster

APA İçier, S., Fındık, F., Göksungur, Y., & Kaplan Türköz, B. (2026). Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein. Türk Doğa ve Fen Dergisi, 15(1), 58-66. https://doi.org/10.46810/tdfd.1752790
AMA 1.İçier S, Fındık F, Göksungur Y, Kaplan Türköz B. Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein. TDFD. 2026;15(1):58-66. doi:10.46810/tdfd.1752790
Chicago İçier, Seyhan, Fatma Fındık, Yekta Göksungur, ve Burcu Kaplan Türköz. 2026. “Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein”. Türk Doğa ve Fen Dergisi 15 (1): 58-66. https://doi.org/10.46810/tdfd.1752790.
EndNote İçier S, Fındık F, Göksungur Y, Kaplan Türköz B (01 Mart 2026) Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein. Türk Doğa ve Fen Dergisi 15 1 58–66.
IEEE [1]S. İçier, F. Fındık, Y. Göksungur, ve B. Kaplan Türköz, “Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein”, TDFD, c. 15, sy 1, ss. 58–66, Mar. 2026, doi: 10.46810/tdfd.1752790.
ISNAD İçier, Seyhan - Fındık, Fatma - Göksungur, Yekta - Kaplan Türköz, Burcu. “Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein”. Türk Doğa ve Fen Dergisi 15/1 (01 Mart 2026): 58-66. https://doi.org/10.46810/tdfd.1752790.
JAMA 1.İçier S, Fındık F, Göksungur Y, Kaplan Türköz B. Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein. TDFD. 2026;15:58–66.
MLA İçier, Seyhan, vd. “Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein”. Türk Doğa ve Fen Dergisi, c. 15, sy 1, Mart 2026, ss. 58-66, doi:10.46810/tdfd.1752790.
Vancouver 1.Seyhan İçier, Fatma Fındık, Yekta Göksungur, Burcu Kaplan Türköz. Validation of whey powder as an inducer for recombinant protein expression from E.coli using GFP as a reporter protein. TDFD. 01 Mart 2026;15(1):58-66. doi:10.46810/tdfd.1752790