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Production of amylases from Coprinus comatus under submerged culture using wheat-milling by-products: Optimization, kinetic parameters, partial purification and characterization
- Frantz, Suélen Caroline, Paludo, Luana Cristina, Stutz, Herta, Spier, Michele Rigon
- Biocatalysis and agricultural biotechnology 2019 v.17 pp. 82-92
- Coprinus comatus, alpha-amylase, calcium, culture media, dipotassium hydrogen phosphate, enzyme activity, experimental design, flour, glucan 1,4-alpha-glucosidase, industrial applications, milling byproducts, pH, starch, temperature, urea, wheat
- Amylases are enzymes that hydrolyze starch, and are mostly produced from microbial sources. This study aimed to evaluate the potential of utilization of wheat milling by-products for the production of amylases by a macromycete strain, Coprinus comatus, recently discovered as a potential producer of amylases under submerged culture. The best results of enzymatic activity (6.07 U mL−1 for α-amylase and 0.64 U mL−1 for glucoamylases) were obtained in a medium containing 10 g L−1 of low-grade flour (LGF) as substrate. The composition of the culture medium was optimized by experimental designs. The enzymatic activities obtained in the optimized culture medium (105 g L−1 LGF, 3.76 g L−1 urea and 7.52 g L−1 K2HPO4) were 59.40 ± 0.56 U mL−1 and 12.65 ± 0.09 U mL−1 for α-amylases and glucoamylases, respectively, representing an increase of 9.78 and 19.77 fold in the initial enzymatic production. High productivity (0.59 U mL−1 h−1 for α-amylase and 0.11 U mL−1 h−1 for glucoamylase) and product formation yield on substrate consumed (814.28 U gds−1 for α-amylase and 231.24 U gds−1 for glucoamylase) were obtained in the kinetic study. These results show that the use of LGF medium represents a low-cost alternative for the production of amylases by the species studied. The characterization of the partially purified enzymes showed optimal activity at temperature of 50 °C and pH 5.0, in addition to the independence of the calcium ion, a desirable characteristic for amylases produced for industrial application.