Isolation of a novel alkaline-induced laccase from Flammulina velutipes and its application for hair coloring

• Published in: Journal of bioscience and bioengineering 2012-05, Vol.113 (5), p.575-579
• Main Authors: (Otsuka) Saito, Kaori, Ikeda, Ryuzo, Endo, Katsunori, Tsujino, Yoshio, Takagi, Masahiro, Tamiya, Eiichi
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Biological and medical sciences; Biotechnology; chromatography; Chromatography, Ion Exchange; Color; Dye; Electrophoresis, Polyacrylamide Gel; Enzyme induction; Flammulina - enzymology; Flammulina velutipes; Fundamental and applied biological sciences. Psychology; Hair - chemistry; Hair coloring; Hair Preparations - chemistry; Humans; hydrogen peroxide; Hydrogen-Ion Concentration; ion exchange; isozymes; Laccase; Laccase - chemistry; Laccase - isolation & purification; Laccase - metabolism; Optimum pH; oxidation; Oxidation-Reduction; polyacrylamide gel electrophoresis; white-rot fungi
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1016/j.jbiosc.2012.01.001

Laccase is a member of the multi-copper oxidase family and a promising for hair coloring. In this study, we isolated a novel alkaline-induced laccase from the white-rot fungus Flammulina velutipes and studied the possibility to apply the enzyme for hair coloring. Laccase activity detected in the culture supernatant of F. velutipes was found to significantly increase when exchanging the medium to laccase inducing one whose pH was adjusted to 9.0. Three isozymes were detected by activity staining on non-denaturing SDS-PAGE. The major isozyme, Flac1, was purified from the culture supernatant after being induced at pH 9.0 by ion-exchange column chromatography. The N-terminal peptide sequence of Flac1 was determined, revealing clear homology with laccases from other white-rot fungi. Optimum pH of oxidation was found to be around pH 5.0−6.5 regardless of several different substrates used. Oxidation activities of Flac1 to several hair dye agents as substrate showed the higher activity at pH 6.5 than that at pH 9.0. Oxidation activity was also detected at pH 9.0 which was suitable for hair coloring. When the purified Flac1 was applied for hair coloring system without using hydrogen peroxide, effective coloring was observed at the protein amount of 0.25mg/1g of hair used. These results indicated that this alkaline-induced novel laccase isolated from the culture supernatant of F. velutipes might be a useful enzyme for hair color.