Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production

Belma Şenol; Özlem Kaplan; Rizvan İmamoğlu; İsa Gökçe

doi:10.31015/jaefs.2021.3.5

EN

Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production

Abstract

Gluten is a protein group found in wheat, barley, rye, and oats, known as cereals. When this vegetable protein is introduced into the body, celiac disease can occurs. The use of bacterial and fungal oligopeptidase to ensure the cleavage of gluten into non-toxic fragments are considered a promising alternative for celiac disease. In this study, the Aspergillus niger Prolyl EndoPeptidase (AN-PEP) enzyme was cloned into pET22b vector and recombinantly produced in BL21 (DE3) pLysE cells. PEP enzyme expressed as inclusion body and was recovered by refolding. And N-terminal His-tagged recombinant protein was purified by nickel affinity chromatography. 280 mg AN-PEP enzyme from 1L bacterial culture was purified at very high yield, and this protein was 90% purity. As a result; It has been determined that the recombinantly produced PEP enzyme can digest gluten. This study shows that recombinantly produced AN-PEP (rAN-PEP) has great potential to use in the production processes of gluten-free foods.

Keywords

Gluten, Celiac disease, Aspergillus niger

Supporting Institution

Tokat Gaziosmanpaşa University Research Fund under

Project Number

2018/32.

Thanks

This work was supported by the Tokat Gaziosmanpaşa University Research Fund under Grant number:2018/32.

References

Babaeipour, V., Abbas, M. P., Sahebnazar, Z., & Alizadeh, R. (2010). Enhancement of human granulocyte-colony stimulating factor production in recombinant E. coli using batch cultivation. Bioprocess Biosyst Eng, 33(5), 591-598. Doi: https://doi.org/10.1007/s00449-009-0380-3
Ciacci, C., Ciclitira, P., Hadjivassiliou, M., Kaukinen, K., Ludvigsson, J. F., McGough, N., Sanders, D. S., Woodward, J., Leonard, J. N., & Swift, G. L. (2015). The gluten-free diet and its current application in coeliac disease and dermatitis herpetiformis. United European Gastroenterol J, 3(2), 121-135.Doi: https://doi.org/10.1177/2050640614559263
Comino, I., Fernandez-Banares, F., Esteve, M., Ortigosa, L., Castillejo, G., Fambuena, B., Ribes-Koninckx, C., Sierra, C., Rodriguez-Herrera, A., Salazar, J. C., Caunedo, A., Marugan-Miguelsanz, J. M., Garrote, J. A., Vivas, S., Lo Iacono, O., Nunez, A., Vaquero, L., Vegas, A. M., Crespo, L., Fernandez-Salazar, L., Arranz, E., Jimenez-Garcia, V. A., Antonio Montes-Cano, M., Espin, B., Galera, A., Valverde, J., Giron, F. J., Bolonio, M., Millan, A., Cerezo, F. M. (2016). Fecal Gluten Peptides Reveal Limitations of Serological Tests and Food Questionnaires for Monitoring Gluten-Free Diet in Celiac Disease Patients. Am J Gastroenterol, 111(10), 1456-1465. Doi: https://doi.org/10.1038/ajg.2016.439
Di Sabatino, A., & Corazza, G. R. (2009). Coeliac disease. Lancet, 373(9673), 1480-1493. Doi: https://doi.org/10.1016/S0140-6736(09)60254-3
Edens, L., Dekker, P., van der Hoeven, R., Deen, F., de Roos, A., & Floris, R. (2005). Extracellular prolyl endoprotease from Aspergillus niger and its use in the debittering of protein hydrolysates. J Agric Food Chem, 53(20), 7950-7957. Doi: https://doi.org/10.1021/jf050652c
Ehren, J., Moron, B., Martin, E., Bethune, M. T., Gray, G. M., & Khosla, C. (2009). A food-grade enzyme preparation with modest gluten detoxification properties. PLoS One, 4(7), e6313. Doi: https://doi.org/10.1371/journal.pone.0006313
Elli, L., Branchi, F., Tomba, C., Villalta, D., Norsa, L., Ferretti, F., Roncoroni, L., & Bardella, M. T. (2015). Diagnosis of gluten related disorders: Celiac disease, wheat allergy and non-celiac gluten sensitivity. World J Gastroenterol, 21(23), 7110-7119. Doi: https://doi.org/10.3748/wjg.v21.i23.7110
Gass, J., Bethune, M. T., Siegel, M., Spencer, A., & Khosla, C. (2007). Combination enzyme therapy for gastric digestion of dietary gluten in patients with celiac sprue. Gastroenterology, 133(2), 472-480. Doi: https://doi.org/10.1053/j.gastro.2007.05.028
Gessendorfer, B., Hartmann, G., Wieser, H., Koehler, P. (2011). Determination of celiac disease-specific peptidase activity of germinated cereals. European Food Research and Technology, 232, 205–209. Doi: https://doi.org/10.1007/s00217-010-1375-7
Helmerhorst, E. J., Zamakhchari, M., Schuppan, D., & Oppenheim, F. G. (2010, Oct 11). Discovery of a novel and rich source of gluten-degrading microbial enzymes in the oral cavity. PLoS One, 5(10), e13264. Doi: https://doi.org/10.1371/journal.pone.0013264

Jiang, B., Wang, M., Wang, X., Wu, S., Li, D., Liu, C., Feng, Z., & Li, J. (2021). Effective separation of prolyl endopeptidase from Aspergillus Niger by aqueous two phase system and its characterization and application. Int J Biol Macromol, 169, 384-395. Doi: https://doi.org/10.1016/j.ijbiomac.2020.12.120
Kaplan, Ö., İmamoğlu, R., Şahingöz, İ., Gökçe, İ. (2021). Recombinant production of Thermus aquaticus single-strand binding protein for usage as PCR enhancer. Int. Adv. Res. Eng. J., 5(1), 42-46. Doi: https://doi.org/10.35860/iarej.766741
Krishnareddy, S., Stier, K., Recanati, M., Lebwohl, B., & Green, P. H. (2017). Commercially available glutenases: a potential hazard in coeliac disease. Therap Adv Gastroenterol, 10(6), 473-481. Doi: https://doi.org/10.1177/1756283X17690991
Kuduğ, H., Ataman, B., İmamoğlu, R., Düzgün, D., Gökçe, İ. (2019). Production of red fluorescent protein (mCherry) in an inducible E. coli expression system in a bioreactor, purification and characterization. Int. Adv. Res. Eng. J., 3(1), 20-25. Retrieved from https://dergipark.org.tr/en/pub/iarej/issue/44303/429547
Lopez, M., & Edens, L. (2005). Effective prevention of chill-haze in beer using an acid proline-specific endoprotease from Aspergillus niger. J Agric Food Chem, 53(20), 7944-7949. Doi: https://doi.org/10.1021/jf0506535
Loponen, J., Kanerva, P., Zhang, C., Sontag-Strohm, T., Salovaara, H., & Ganzle, M. G. (2009). Prolamin hydrolysis and pentosan solubilization in germinated-rye sourdoughs determined by chromatographic and immunological methods. J Agric Food Chem, 57(2), 746-753. Doi: https://doi.org/10.1021/jf803243w
Melissis, S. C., Papageorgiou, A. C., Labrou, N. E., & Clonis, Y. D. (2010). Purification of M-MLVH- RT on a 9-aminoethyladenine-(1,6-diamine-hexane)-triazine selected from a combinatorial library of dNTP-mimetic ligands. J Chromatogr Sci, 48(6), 496-502. Doi: https://doi.org/10.1093/chromsci/48.6.496
Moreno, M. L., Cebolla, A., Munoz-Suano, A., Carrillo-Carrion, C., Comino, I., Pizarro, A., Leon, F., Rodriguez-Herrera, A., & Sousa, C. (2017). Detection of gluten immunogenic peptides in the urine of patients with coeliac disease reveals transgressions in the gluten-free diet and incomplete mucosal healing. Gut, 66(2), 250-257. Doi: https://doi.org/10.1136/gutjnl-2015-310148
Ortiz, C., Valenzuela, R., & Lucero, A. Y. (2017). Celiac disease, non celiac gluten sensitivity and wheat allergy: comparison of 3 different diseases triggered by the same food. Rev Chil Pediatr, 88(3), 417-423. Doi: https://doi.org/10.4067/S0370-41062017000300017
Palmer, I., & Wingfield, P. T. (2012). Preparation and extraction of insoluble (inclusion-body) proteins from Escherichia coli. Curr Protoc Protein Sci, Chapter 6, Unit6 3. Doi: https://doi.org/10.1002/0471140864.ps0603s70
Rodrigues, M., Yonamine, G. H., & Fernandes Satiro, C. A. (2018). Rate and determinants of non-adherence to a gluten-free diet and nutritional status assessment in children and adolescents with celiac disease in a tertiary Brazilian referral center: a cross-sectional and retrospective study. BMC Gastroenterol, 18(1), 15. Doi: https://doi.org/10.1186/s12876-018-0740-z
Schwalb, T., Wieser, H., Koehler, P. (2012). Studies on the gluten-specificpeptidase activity of germinated grains from different cereal species and cultivars. European Food Research and Technology, 235,1161–1170.
Sebela, M., Rehulka, P., Kabrt, J., Rehulkova, H., Ozdian, T., Raus, M., Franc, V., & Chmelik, J. (2009). Identification of N-glycosylation in prolyl endoprotease from Aspergillus niger and evaluation of the enzyme for its possible application in proteomics. J Mass Spectrom, 44(11), 1587-1595. Doi: https://doi.org/10.1002/jms.1667
Shan, L., Marti, T., Sollid, L. M., Gray, G. M., & Khosla, C. (2004). Comparative biochemical analysis of three bacterial prolyl endopeptidases: implications for coeliac sprue. Biochem J, 383(Pt 2), 311-318. Doi: https://doi.org/10.1042/BJ20040907
Siegel, M., Bethune, M. T., Gass, J., Ehren, J., Xia, J., Johannsen, A., Stuge, T. B., Gray, G. M., Lee, P. P., & Khosla, C. (2006). Rational design of combination enzyme therapy for celiac sprue. Chem Biol, 13(6), 649-658. Doi: https://doi.org/10.1016/j.chembiol.2006.04.009
Stepniak, D., Spaenij-Dekking, L., Mitea, C., Moester, M., de Ru, A., Baak-Pablo, R., van Veelen, P., Edens, L., & Koning, F. (2006). Highly efficient gluten degradation with a newly identified prolyl endoprotease: implications for celiac disease. Am J Physiol Gastrointest Liver Physiol, 291(4), G621-629. Doi: https://doi.org/10.1152/ajpgi.00034.2006
Tye-Din, J. A., Anderson, R. P., Ffrench, R. A., Brown, G. J., Hodsman, P., Siegel, M., Botwick, W., & Shreeniwas, R. (2010). The effects of ALV003 pre-digestion of gluten on immune response and symptoms in celiac disease in vivo. Clin Immunol, 134(3), 289-295. Doi: https://doi.org/10.1016/j.clim.2009.11.001
Vallejo, L. F., & Rinas, U. (2004). Strategies for the recovery of active proteins through refolding of bacterial inclusion body proteins. Microb Cell Fact, 3(1), 11. Doi: https://doi.org/10.1186/1475-2859-3-11
Walter, T., Wieser, H., Koehler, P. (2015). Degradation of gluten in rye sourdough products by means of a proline-specific peptidase. . European Food Research and Technology, 240, 517–524. Doi: https://doi.org/10.1007/s00217-014-2350-5
Wei, G., Helmerhorst, E. J., Darwish, G., Blumenkranz, G., & Schuppan, D. (2020). Gluten Degrading Enzymes for Treatment of Celiac Disease. Nutrients, 12(7). Doi: https://doi.org/10.3390/nu12072095
Wei, G., Tian, N., Valery, A. C., Zhong, Y., Schuppan, D., & Helmerhorst, E. J. (2015). Identification of Pseudolysin (lasB) as an Aciduric Gluten-Degrading Enzyme with High Therapeutic Potential for Celiac Disease. Am J Gastroenterol, 110(6), 899-908. Doi: https://doi.org/10.1038/ajg.2015.97
Zamakhchari, M., Wei, G., Dewhirst, F., Lee, J., Schuppan, D., Oppenheim, F. G., & Helmerhorst, E. J. (2011). Identification of Rothia bacteria as gluten-degrading natural colonizers of the upper gastro-intestinal tract. PLoS One, 6(9), e24455. Doi: https://doi.org/10.1371/journal.pone.0024455

Details

Primary Language

English

Subjects

Food Engineering

Journal Section

Research Article

Authors

Belma Şenol This is me
0000-0003-3021-0087
Türkiye

Özlem Kaplan
0000-0002-3052-4556
Türkiye

Rizvan İmamoğlu
0000-0002-6306-4760
Türkiye

İsa Gökçe ^*
0000-0002-5023-9947
Türkiye

Publication Date

September 15, 2021

Submission Date

February 10, 2021

Acceptance Date

June 4, 2021

Published in Issue

Year 2021 Volume: 5 Number: 3

DOI

https://doi.org/10.31015/jaefs.2021.3.5

IZ

https://izlik.org/JA93DK27DT

APA

Şenol, B., Kaplan, Ö., İmamoğlu, R., & Gökçe, İ. (2021). Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production. International Journal of Agriculture Environment and Food Sciences, 5(3), 287-293. https://doi.org/10.31015/jaefs.2021.3.5

AMA

1.Şenol B, Kaplan Ö, İmamoğlu R, Gökçe İ. Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production. int. j. agric. environ. food sci. 2021;5(3):287-293. doi:10.31015/jaefs.2021.3.5

Chicago

Şenol, Belma, Özlem Kaplan, Rizvan İmamoğlu, and İsa Gökçe. 2021. “Recombinant Production and Characterization of Aspergillus Niger Prolyl Endopeptidase Enzyme for Gluten-Free Food Production”. International Journal of Agriculture Environment and Food Sciences 5 (3): 287-93. https://doi.org/10.31015/jaefs.2021.3.5.

EndNote

Şenol B, Kaplan Ö, İmamoğlu R, Gökçe İ (September 1, 2021) Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production. International Journal of Agriculture Environment and Food Sciences 5 3 287–293.

IEEE

[1]B. Şenol, Ö. Kaplan, R. İmamoğlu, and İ. Gökçe, “Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production”, int. j. agric. environ. food sci., vol. 5, no. 3, pp. 287–293, Sept. 2021, doi: 10.31015/jaefs.2021.3.5.

ISNAD

Şenol, Belma - Kaplan, Özlem - İmamoğlu, Rizvan - Gökçe, İsa. “Recombinant Production and Characterization of Aspergillus Niger Prolyl Endopeptidase Enzyme for Gluten-Free Food Production”. International Journal of Agriculture Environment and Food Sciences 5/3 (September 1, 2021): 287-293. https://doi.org/10.31015/jaefs.2021.3.5.

JAMA

1.Şenol B, Kaplan Ö, İmamoğlu R, Gökçe İ. Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production. int. j. agric. environ. food sci. 2021;5:287–293.

MLA

Şenol, Belma, et al. “Recombinant Production and Characterization of Aspergillus Niger Prolyl Endopeptidase Enzyme for Gluten-Free Food Production”. International Journal of Agriculture Environment and Food Sciences, vol. 5, no. 3, Sept. 2021, pp. 287-93, doi:10.31015/jaefs.2021.3.5.

Vancouver

1.Belma Şenol, Özlem Kaplan, Rizvan İmamoğlu, İsa Gökçe. Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production. int. j. agric. environ. food sci. 2021 Sep. 1;5(3):287-93. doi:10.31015/jaefs.2021.3.5

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Abstract

Recombinant production and characterization of Aspergillus niger prolyl endopeptidase enzyme for gluten-free food production

Abstract

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Supporting Institution

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Thanks

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Acceptance Date

Published in Issue

DOI

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Cited By

High-level production of Aspergillus niger prolyl endopeptidase from agricultural residue and its application in beer brewing

Soluble Expression of Human Granulocyte Colony Stimulating Factor (hG-CSF) in Escherichia coli Expression System

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