Purification and characterization of azoreductase

• Main Authors: Hu Chih Hsien, 胡志賢
• Other Authors: Lin Ping Jei
• Format: Others
• Language: zh-TW
• Published: 2000
• Online Access: http://ndltd.ncl.edu.tw/handle/44192990352336981285

碩士 === 逢甲大學 === 化學工程學系 === 88 === In the past decades，the applications of biotechnology was developed in many asects to reduce environmental pollutants. The contained azo reductase of Pseudomonas luteola was found the ability to decolarize the reactive azo dyes. This study investigated the purifications and the kinetics of the enzyme. The enzyme of a crude cell extract was purified in two stages. First，it was eluted by means of either cation or anion exchange chromatography. The specific activities of the enzyme were 25.574 nM dye/min/mg protein and 27.733 nM dye/min/ mg protein after the purifications by cation and anion exchange chromatography，respectively. The value was approximately 40 times about that of the crude cell extract (0.663 nM dye/min/mg protein). Second，the purified enzyme from the first stage was introduced to the gel permeation chromatographic columns for further purification. The specific activity of theenzyme increased to 57.88 nM dye/min/mg protein，approximately 80-fold high than that of the crude cell extract. In the kinetics study，the Michalis-Menten constants of the crude cell extract enzyme and the purified enzyme were 11.656μM and 6.167μM；respectively.