Determination of Cell Permeabilization and Beta-Galactosidase Extraction from <i>Aspergillus oryzae</i> CCT 0977 Grown in Cheese Whey

Viana, Caroline dos Santos; Pedrinho, Denise Renata; Morioka, Luiz Rodrigo Ito; Suguimoto, Hélio Hiroshi

doi:10.1155/2018/1367434

Cited by 10 publications

(6 citation statements)

References 15 publications

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“…1c). These values of solvent, time and temperature were lower than those used by Viana et al 19 . for the β‐galactosidase extraction of Aspergillus oryzae CCT 0977 with 5% solvent, overnight at 45 °C.…”

Section: Resultsmentioning

confidence: 69%

Extraction of β‐galactosidase from Saccharomyces fragilisIZ 275 grown in cheese whey

Setti

Bosso

Morioka

et al. 2022

J of Chemical Tech & Biotech

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BACKGROUND The enzyme β‐galactosidase (EC 3.2.1.2.3) obtained from yeast is produced intracellularly. Cell extraction consists of the treatment of a cell suspension with chemical agents, thus obtaining the enzyme with biological activity. After obtaining the extracted enzyme, it is concentrated by ultrafiltration and dried. Lyophilization can be used in this step. The objective of this work was to study the production and extraction of β‐galactosidase from Saccharomyces fragilis IZ 275 using lactose from cheese whey. RESULTS Under optimum extraction conditions, 2% (v/v) chloroform, 37 °C for 6 h of extraction, 35% of lactose hydrolysis was achieved using 1% (v/v) β‐galactosidase during 10 h of hydrolysis. Extracted enzyme activity resulted in 81% lactose hydrolysis using 12% (v/v) enzyme at pH 6.0 and 37 °C. Enzymatic kinetics were evaluated using 12% of extracted enzyme in 5 h of hydrolysis reaction, and 100% lactose hydrolysis was obtained. The enzyme activity after extraction, concentration and lyophilization was 100%, 60% and 45.3% of lactose hydrolysis, respectively, after 5 h of hydrolysis reaction at 37 °C and pH 6.0. CONCLUSION The yeast S. fragilis IZ 275 grown in cheese whey produces intracellular β‐galactosidase and its extraction is efficient with the use of 2% chloroform at 37 °C for 6 h, allowing its use for industrial and pharmaceutical purposes. The extracted enzyme must be used at 37 °C and pH 6.0 at a concentration of 12% (v/v) of enzyme to obtain the total hydrolysis of lactose in 5 h of reaction. © 2022 Society of Chemical Industry (SCI).

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Section: Resultsmentioning

confidence: 69%

Extraction of β‐galactosidase from Saccharomyces fragilisIZ 275 grown in cheese whey

Setti

Bosso

Morioka

et al. 2022

J of Chemical Tech & Biotech

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“…Therefore, it can be naturally present in the air and in the environment where the cheeses are ripened, since cheese ripening rooms are usually located next to crops. Aspergillus oryzae produces the Beta-Galactosidase enzyme (Viana, et. al, 2018), which is used to break down lactose.…”

Section: Discussionmentioning

confidence: 99%

Fungal Community and Physicochemical Profiles of Ripened Cheeses

Aragão¹,

Evangelista²,

Passamani³

et al. 2021

Preprint

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Ripened cheeses are traditionally produced and consumed worldwide. Canastra’s Minas artisanal cheese (QMA) is a protected geographical indication (PGI) traditional ripened cheeses. The influence of fungi on the cheese ripening process is of great importance. This study aimed to apply culture-dependent and -independent methods to determine the mycobiota of QMA produced in the Canastra region, as well as to determine its physicochemical characteristics. Samples from different producers were collected in the cities of São Roque de Minas and Piumhi (MG). Illumina-based amplicon sequencing, and Matrix Assisted Laser Desorption Ionization Time-of-Flight - (MALDI-TOF) Mass Spectrometry (MS) methods were used. The physicochemical analysis showed that the QMA had a moisture content between 18.24% and 21%, fat content between 20.5% and 40%, sodium chloride percentage around 0.9%, and pH of 5.5 to 5.3. The population of fungi ranged between 6.3 and 8 log CFU/g. Fusarium sp., Geotrichum candidum, Paecilomyces sp., Trichosporon coremiiforme, Candida catenulata, Aspergillus sp., Trichosporon japonicum, A. oryzae, Kluyveromyces, Torulaspora, and Debaryomyces were the most prevalent fungi. A. ochraceus potentially mycotoxin-producing was detected. Promising species that can contribute to the product quality were identified as Geotrichum candidum and Candida catenulate. However, future studies should evaluate the ability of A. ochraceus to produce mycotoxin in cheese, as well as control the growth of certain species of fungi during ripening, resulting in a safe and high quality product.

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“…However, because of the low amount of excreted redox‐active species and their slow excretion rates, the electrochemical signal in most fungi is very low. Nevertheless, the amount of excreted redox‐active species, especially in Tris buffer, can be increased by permeabilizing the cell membrane without the use of harsh permeabilization processes [28, 34–35]. Moreover, an electrochemical signal of a redox‐active species can be amplified using electrochemical redox cycling when its oxidized and reduced forms are stable.…”

Section: Introductionmentioning

confidence: 99%

Simple and rapid detection of Alternaria alternata using an excreted redox‐active species

et al. 2023

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Airborne fungi pose a serious threat to public health. Alternaria alternata (A. alternata) is a fungus that has been associated with the development of asthma. Detection using redox‐active species excreted from fungi is an effective method for a simple electrochemical fungal biosensor. The achievable electrochemical signal in most fungi, however, is exceptionally low because of the low amount of excreted redox‐active species and their slow excretion rates. Herein, we report that A. alternata excretes an exceptionally large amount of a redox‐active species that can be used for sensitive and selective detection of A. alternata. The excretion rate is enhanced in Tris buffer, and the electrochemical‐chemical redox cycling involving excreted redox‐active species significantly increases the electrochemical signals. Only A. alternata among five common airborne fungi provides large electrochemical signals, which allows selective detection of A. alternata. The calculated detection limit for A. alternata is ~20 spores/mL with an incubation period of 10 min, indicating that the detection method is highly sensitive and rapid. The detection method does not require complicated procedures or harsh pretreatment and is optimal for point‐of‐care testing of A. alternata.

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Determination of Cell Permeabilization and Beta-Galactosidase Extraction from Aspergillus oryzae CCT 0977 Grown in Cheese Whey

Cited by 10 publications

References 15 publications

Extraction of β‐galactosidase from Saccharomyces fragilisIZ 275 grown in cheese whey

Extraction of β‐galactosidase from Saccharomyces fragilisIZ 275 grown in cheese whey

Fungal Community and Physicochemical Profiles of Ripened Cheeses

Simple and rapid detection of Alternaria alternata using an excreted redox‐active species

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