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| Dong,Bing Xue; Zhang,Yu Hong; Li,Gang; Fang,Jin Tao; Zheng,Ji Huan; Mao,Run Qian. |
| ABSTRACT To seek a simple, rapid and sensitive Coprinus cinereus Peroxidase (CIP) activity assay, a convenient one-factor-at-a-time (OFAT) method and a response surface methodology (RSM) were used. The recombinant CIP expressed in Pichia pastoris was purified with the Ni-NTA spin column. Based on the results of catalytic efficiency (kcat/Km) analysis, 2,2'-azinobis (ethylbenzthiazoline -6-sulfonate) (ABTS) was selected as the optimal enzyme substrate. Results of the OFAT method showed that enzymatic reaction performed in 0.1 mol/L sodium acetate (pH 5.0) buffer in a 200-µl reaction mixture containing 0.5 mmol/L ABTS, 10 mmol/L hydrogen peroxide (H2O2), 49.7 ng CIP at 25°C gave an average CIP activity of 88 U/mL. The ABTS and H2O2 concentrations were then... |
| Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Enzymatic activity assay; One-factor-at-a-time (OFAT) method; Response surface methodology (RSM); Coprinus cinereus peroxidase (CIP). |
| Ano: 2018 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100421 |
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