[Properties of a triphenylmethane dyes decolorization enzyme TpmD from Aeromonas hydrophila strain DN322].
A novel bacterial decolorization enzyme for triphenylmethane dyes from Aeromonas hydrophila strain DN322 was purified, named TpmD. The purified enzyme catalyzes the decolorization of several triphenylmethane dyes, i.e., crystal violet, basic fuchsin, brilliant green and malachite green. The enzyme was identified by the clear transparent band development of zymogram stained with crystal violet, basic fuchsin, brilliant green and malachite green after polyacrylamide gel electrophoresis (PAGE) respectively. The decolorization enzyme was enzymologically characterized. The results showed that the molecular weight of TpmD is 29.4kDa and its isoelectric point (pI) is 5.6. The maximal activity of TpmD for above four triphenylmethane dyes was observed at 50 degrees C - 55 degrees C and pH 7.4 - 8.0. The temperature for losing half of the activity (t1/2) within 4h is 62 degrees C. The activities of decolorization enzyme are relatively stable at pH range of 5.5 - 9.0. The K(m) and V(max) of TpmD for decolorizing crystal violet, basic fuchsin, brilliant green and malachite green are 24.3, 40.6, 54.2, 68.5 micromol/L respectively, V(max) are 19.6, 74.1, 82.8, 115.6 micromol x L(-1) x s(-1) respectively. Both NADH/NADPH and molecular oxygen are necessary for the enzyme to decolorize triphenylmethane dyes, indicate the enzyme is an NADH/NADPH-dependent oxygenase.
Ren
Sui-Zhou
SZ
Guangdong Institute of Microbiology, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangzhou 510070, China. rensz@163.com
Guo
Jun
J
Wang
Ya-Li
YL
Cen
Ying-Hua
YH
Sun
Guo-Ping
GP
D016980:Aeromonas hydrophila / Q000201:enzymology*; D003116:Color; D004396:Coloring Agents / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D009243:NAD / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D013696:Temperature; D014320:Triphenylmethyl Compounds / Q000378:metabolism*
D004396:Coloring Agents; D014320:Triphenylmethyl Compounds; D009243:NAD; C046945:triphenylmethane; D010100:Oxygen