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The psychrotrophic yeast Sporobolomyces roseus LOCK 1119 as a source of a highly active aspartic protease for the in vitro production of antioxidant peptides
Author(s) -
Białkowska Aneta M.,
Krysiak Joanna,
Florczak Tomasz,
Szulczewska Katarzyna M.,
Wanarska Marta,
Turkiewicz Marianna
Publication year - 2018
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.1656
Subject(s) - pepstatin , protease , biochemistry , enzyme , biology , yeast extract , pmsf , enzyme assay , fast protein liquid chromatography , yeast , fermentation
A psychrotrophic yeast strain producing a cold‐adapted protease at low temperature was classified as Sporobolomyces roseus . In standard YPG medium, S. roseus LOCK 1119 synthesized an extracellular protease with an activity of approximately 560 U/L. Optimization of medium composition and process temperature considerably enhanced enzyme biosynthesis; an approximate 70% increase in activity (2060 U/L). The native enzyme was purified to homogeneity by cation exchange chromatography followed by a size exclusion step, resulting in a 103‐fold increase in specific activity (660 U/mg) with 25% recovery. The enzyme displayed 10%–30% of its maximum activity at 0–25 °C, with the optimum temperature being 50°C. Protease G8 was strongly inactivated by pepstatin A, an aspartic protease inhibitor. The enzyme was used to hydrolyze four natural substrates, and their antioxidant activities were evaluated against 1,1‐diphenyl‐2‐picrylhydrazyl. The highest antioxidant activity (69%) was recorded for beef casein.