Saccharomyces fragilis IZ 275 IN CHEESE WHEY CELL PERMEABILIZATION USING DIFFERENT ORGANIC SOLVENTS
Keywords:β-galactosidase; disruption cell yeast; scanning electron microscopy; biotechnological process; green solvent.
The aim of this study was to verify the efficacy of different organic solvents, in the permeabilization of Saccharomyces fragilis IZ 275, by using a Central Composite Rotational Design (CCRD) 23 and Response Surface Methodology (RSM). Furthermore, we aimed to evaluate the effectiveness of the permeabilization process by monitoring lactose hydrolysis and obtaining images of non-permeabilized and permeabilized cells by Scanning Electron Microscopy (SEM). The yeast S. fragilis IZ 275 was grown in a fermentation medium composed of cheese whey, and the permeabilized cells was estimated by β-galactosidase activity. The response surface methodology was used as it is an efficient tool to optimize the permeabilization process as well as to identify the organic solvent which was most effective for this process. Our results show that the concentration and type of organic solvent, as well as permeabilization temperature and time influence the cells permeabilization process.. Considering the experimental results, the best conditions when using chloroform are a concentration of 4 % at 25 ºC during 20 min with 81.03 % lactose hydrolysis. In this study, we found that the use of ethanol for cellular permeabilization lead to obtaining β-galactosidase enzyme, a process which can be used in a large scale by the food industry, being a cheaper and more environmentally safe way of obtaining this enzyme.
Anisha, G.S. (2017). β-galactosidases. Current Developments in Biotechnology and Bioengineering: Production, Isolation and Purification of Industrial Products, 17, 395- 421.
Ballus, C.A., Meinhart, A.D., Bruns, R.E., & Godoy, H.T. (2011). Use of multivariate statistical techniques to optimize the simultaneous separation of 13 phenolic compounds from extra-virgin olive oil by capillary electrophoresis. Talanta, 83, 1181–1187.
Becker, J.M., Caldwell, G.A., & Zachgo, E.A. (1996). Determination of β-Galactosidase in Permeabilized Yeast Cells. In: Becker, J M.; Caldwell, G. A.; Zachgo, E. A. Biotechnology: a technology course. 2. ed. Academic Press, 131-134.
Bruns, R.E., Scarminio, I.S., & Barros Neto, B. (2006). Statistical Design-Chemometrics. In: Data Handling in Science and Technology, vol. 25, Elsevier, Amsterdam.
Chemat, F., Vian, M.A., & Cravotto, G. (2012). Green Extraction of Natural Products: Concept and Principles. International Journal Molecular Science, 13, 8615-8627.
Flores, M.V., Voget, C.E., & Ertola, R.J.J. (1994). Permeabilization of yeast cells (Kluyveromyces lactis) with organic solvents. Enzyme Microbiology Technology. 16, 340–346.
Geciova, J., Bury, & D., Jelen, P. (2002). Methods for disruption of microbial cells for potential use in the dairy industry-a review. International Dairy Journal, 12, 541–553.
Guimarães, P M., Teixeira, J.A., & Domingues, L. (2010). Fermentation of lactose to bio-ethanol by yeasts as part of integrated solutions for the valorisation of cheese whey. Biotechnology Advance, 28, 375-84.
Husain, Q. (2010). Beta galactosidases and their potential applications: a review. Critical Review Biotechnology, 30, 41–62.
Kaur, G., Panesar, P.S., Bera, M.B., & Kumar, H. (2009). Hydrolysis of whey lactose using CTAB-permeabilized yeast cells. Bioprocess Biosyst Eng., 32, 63–67.
Mlichova, Z., & Rosenberg, M. (2006). Current trends of β-galactosidase application in food technology. Journal Food Nutritional Research, 45, p. 47–54.
Nath, A., Verasztó, B., Basak, S., Koris, A., Kovács, & Z. Vatai, G. (2016). Synthesis of lactose-derived nutraceuticals from dairy waste whey – A review. Food Bioprocess Tech., 9, 16–48.
Oak, S.J., & Jha, R. (2018). The effects of probiotics in lactose intolerance: A systematic review, Critical Reviews in Food Science and Nutrition. https://doi.org/10.1080/10408398.2018.1425977.
Panesar, P.S., Panesar, R., Singh, R.S., Kennedy, J.F., & Kumar, H. (2006). Microbial production, immobilization and applications of β-D-galactosidase. Journal of Chemistry Technical Biotechnology, 81, 530-430.
Panesar, P.S., Kumari, S., & Panesar, R. (2013). Biotechnological approaches for the production of prebiotics and their potential applications. Critical Review Biotechnol., 33, 345-64.
Panesar, P.S., Panesar, R., Singh, R.S., & Bera, M.B. (2007). Permeabilization of yeast cells with organic solvents for β-galactosidase activity. Research Journal of Microbiology, 2(1), 34–41.
Panesar, P.S., & Kennedy, J.F. (2012). Biotechnological approaches for the value addition of whey. Critical Reviews in Biotechnology, 32 (4), 327–348.
Prasad, L.N., Ghosh, B.C., Sherkat, F., & Shah, N.P. (2013). Extraction and characterisation of β-galactosidase produced by Bifidobacterium animalis spp. lactis Bb12 and Lactobacillus delbrueckii spp. bulgaricus ATCC 11842 grown in whey. International Food Research Journal, 20, 487-498.
Silveira, T.F., Meinhart, A.D., Souza, T.C., Teixeira Filho, J., & Godoy, H.T. (2016). Phenolic compounds from yerba mate based beverages-A multivariate optimization. Food Chemistry, 190, 1159-1167.
Viana, C.S., Pedrinho, D.R., Morioka, L.R.I., & Suguimoto, H.H. (2018). Determination of cell permeabilization and beta-galactosidase extraction from Aspergillus oryzae CCT 0977 grown in whey cheese. International Journal of Chemical Engineering, 2018. https://doi.org/10.1155/2018/1367434
How to Cite
Copyright (c) 2020 Alessandra Bosso, Adriana Aparecida Bosso Tomal, Lucas Calderiao, Josemeyre Bonifacio Da Silva, Raul Jorge Hernan Castro-Gomez, Hélio Hiroshi Suguimoto
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
1) Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2) Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
3) Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.