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Production, characterization, and immobilization of protease from the yeast Rhodotorula oryzicola
Author(s) -
Oliveira Juliana Mota,
Fernandes Pedro,
Benevides Raquel Guimarães,
Assis Sandra Aparecida
Publication year - 2021
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.2023
Subject(s) - protease , chromatography , chemistry , enzyme , yeast , enzyme assay , size exclusion chromatography , casein , immobilized enzyme , rhodotorula , yeast extract , specific activity , biochemistry , fermentation
The protease was produced extracellularly in submerged fermentation by the yeast Rhodotorula oryzicola using different sources of nitrogen and maximum activity (6.54 × 10 −3 U/mg) was obtained in medium containing 2% casein (w/v). Purification of the protease by gel filtration chromatography resulted in a 3.07‐fold increase of specific protease activity. The optimal pH and temperature for enzyme activity were 6.51 and 63.04 °C, respectively. Incubation in the presence of some salts enhanced enzyme activity, which peaked under 0.01 M BaCl 2 . The enzyme retained about 90% of enzymatic activity at temperatures 50–60 °C. The commercially available enzyme carriers evaluated, silica gel, Celite 545, and chitosan effectively immobilized the protease. The enzyme immobilized in Celite 545 retained 73.53% of the initial activity after 15 reuse cycles. These results are quite promising for large‐scale production and immobilization of protease from R. oryzicola , as the high operational stability of the immobilized enzyme lowers production costs in biotechnological applications that require high enzymatic activity and stability under high temperatures.