Permeabilization of Yeast Cells for β-Galactosidase Activity using Mixture of Organic Solvents: A Response Surface Methodology Approach

Abstract Permeabilization was used for the purpose of transforming the cells of microorganisms into biocatalysts with an enhanced enzyme activity. Baker’s yeast cells were permeabilized with various organic solvents. A high degree of catalase activity was observed upon permeabilization with acetone, chloroform, isopropyl alcohol and ethyl acetate. Response surface methodology was used to model the effect of concentration of isopropyl alcohol, temperature and treatment time on the permeabilization of baker’s yeast cells to maximize the decomposition of H2O2. The optimum operating conditions for permeabilization were observed at 53.7% concentration of isopropyl alcohol, treatment time of 40 min and temperature of 15.6oC. A maximum value of catalase activity was found to be 6.188 U/g wet wt. and was ca. 60 times higher than the catalytic activity of yeast not treated by the permeabilization process.

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Optimization of Parameters Using Response Surface Methodology to Develop a Novel Kefir-Like Functional Beverage from Cheese Whey Enriched with Myrtle Juice

Journal of Chemistry ◽

10.1155/2021/2984470 ◽

2021 ◽

Vol 2021 ◽

pp. 1-13

Author(s):

Sana M’hir ◽

Asma Mejri ◽

Hajer Atrous ◽

Lamia Ayed

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Goodness Of Fit ◽

Raw Material ◽

Yeast Cells ◽

Human Consumption ◽

Average Deviation ◽

Total Polyphenol ◽

Overall Acceptability ◽

Cheese Production

Whey, liquid wastewater from cheese production, is one of the sources of dietary protein and lactose that are still largely unused for human consumption. It is only in recent years that it has aroused the interest of industries and sought as a valuable raw material and thus represents an opportunity for the manufacture of new products. The manufacture of fermented whey drink requires the mixing of whey with fruit juice or an aromatic plant to improve its organoleptic properties and acceptability. Myrtle, an aromatic medicinal plant, known for its health benefits is not well exploited for making dairy products. This is the first report on the development of kefir-myrtle beverage. Three factors were optimized (whey permeates (%), myrtle’s juice (%), and kefir grains as inoculum (%)) using a central composite design with response surface methodology. The analyses showed that the number of lactic acid bacteria (LAB) and yeast cells varied from 5.4 to 9.2 log10 CFU/mL and from 4.3 to 6.2 log10 CFU/mL, respectively. A decrease in pH and an increase in the total polyphenol content and antioxidant activity were observed. The analysis of variance indicated the goodness of fit of the model with R2 from 0.827 to 0.966. The absolute average deviation values of each model were low and ranged from 1.61% to 4.23%. The optimized fermented kefir whey beverage accomplished an overall acceptability of 5.41 (1 to 9 preference scale) and a high number of LAB cells (8.53 log10 CFU/mL). The viability of LAB and yeast cell was maintained at 7.61 and 6.19 log10 CFU/mL, respectively, after 14 days of storage.

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Optimization of permeabilization process of yeast cells for catalase activity using response surface methodology

Biotechnology & Biotechnological Equipment ◽

10.1080/13102818.2014.934986 ◽

2015 ◽

Vol 29 (1) ◽

pp. 72-77 ◽

Cited By ~ 2

Author(s):

Ilona Trawczyńska ◽

Marek Wójcik

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Catalase Activity ◽

Yeast Cells

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Optimization of Arabitol Production by Karyoductant SP-K 7 of Saccharomyces cerevisiae V30 and Pichia stipitis CCY 39501 Using Response Surface Methodology

Polish Journal of Microbiology ◽

10.33073/pjm-2012-039 ◽

2012 ◽

Vol 61 (4) ◽

pp. 291-297 ◽

Cited By ~ 1

Author(s):

MONIKA KORDOWSKA-WIATER ◽

AGNIESZKA KUBIK-KOMAR ◽

ZDZISŁAW TARGOŃSKI

Keyword(s):

Saccharomyces Cerevisiae ◽

Response Surface Methodology ◽

Response Surface ◽

Rotation Speed ◽

Pichia Stipitis ◽

Culture Conditions ◽

Yeast Cells ◽

Experimental Conditions ◽

Batch Cultures ◽

Pharmaceutical Industries

L-arabitol is used in the food and pharmaceutical industries. It can be secreted by genetically modified Saccharomyces cerevisiae carrying the genes responsible for pentose metabolism in yeast cells. The process of the biotransformation of L-arabinose to arabitol is highly dependent on culture conditions. The aim of this investigation was to use statistical response surface methodology (RSM) for optimization of biotransformation of L-arabinose to arabitol by a karyoductant of S. cerevisiae V30 and Pichia stipitis CCY 39501, named SP-K7. Batch cultures of yeast were performed according to a Plackett-Burman design, and three factors, rotation speed, L-arabinose concentration, and temperature, were chosen for a central composite design (CCD) applied in order to optimize the production of the polyol by the karyoductant. On the basis of results obtained using 20 combinations of batch cultures of karyoductant SP-K7, the optimal levels of the factors were determined as: rotation speed 150 rpm, concentration of L-arabinose 32.5 g/l, and temperature 28 degrees C. In such conditions, the predicted concentration of arabitol after two days of incubation of SP-K7 should be 18.367 g/l. The value of R2 = 0.93195 suggested that this model was well-fitted to the experimental data. A verification of the model in experimental conditions confirmed its usefulness.

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Saccharomyces fragilis IZ 275 IN CHEESE WHEY CELL PERMEABILIZATION USING DIFFERENT ORGANIC SOLVENTS

Research Society and Development ◽

10.33448/rsd-v9i9.6952 ◽

2020 ◽

Vol 9 (9) ◽

pp. e113996952

Author(s):

Alessandra Bosso ◽

Adriana Aparecida Bosso Tomal ◽

Lucas Caldeirão ◽

Josemeyre Bonifacio da Silva ◽

Raul Jorge Hernan Castro-Gomez ◽

...

Keyword(s):

Response Surface Methodology ◽

Organic Solvent ◽

Response Surface ◽

Organic Solvents ◽

Large Scale ◽

Cheese Whey ◽

Fermentation Medium ◽

Lactose Hydrolysis ◽

Cell Permeabilization ◽

Permeabilized Cells

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.

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