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<span style="font-size:15.0pt;mso-fareast-font-family:"Times New Roman";mso-ansi-language: EN-GB;mso-fareast-language:EN-US;mso-bidi-language:HI" lang="EN-GB">Purification, characterization and synthetic application of a thermally stable laccase from<i> Hexagonia tenuis</i> MTCC-1119</span>

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Title Purification, characterization and synthetic application of a thermally stable laccase from Hexagonia tenuis MTCC-1119
 
Creator Chaurasia, Pankaj Kumar
Bharati, Shashi Lata
Yadava, Sudha
Yadav, Rama Shanker Singh
 
Subject Laccases
Hexagonia tenuis
Metalloenzymes
Lignolytic enzymes
Cu containing enzymes
 
Description 60-67
A thermally stable
laccase was purified from the culture filtrate of Hexagonia tenuis MTCC-1119. The method involved concentration of
the culture filtrate by ammonium sulphate precipitation and an anion-exchange
chromatography on diethylaminoethyl (DEAE) cellulose. The sodium dodecyl
sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and native
polyacrylamide gel electrophoresis (native-PAGE) both gave single protein
bands, indicating that the enzyme preparation was pure. The molecular mass of
the enzyme determined from SDS-PAGE analysis was 100 kDa. The purification fold
and percentage recovery of the enzyme activity were 12.75 and 30.12%,
respectively. The pH and the
temperature optima were 3.5 and 45○C, respectively. The enzyme was
most stable at pH 4.0 when exposed
for 1 h. Using 2,6-dimethoxyphenol (DMP), 2,2’
[azino-bis-(3-ethylbonzthiazoline-6-sulphonic acid) diammonium salt] (ABTS) and

3,5-dimethoxy-4-hydroxybenzaldehyde azine (syringaldazine) as the substrates,
the Km, kcat and kcat/Km values of the laccase were 80 μM, 2.54 s-1,
3.17 × 104 M-1s-1, 36 μM, 2.54 s-1,
7.05 × 104 M-1s-1 and 87 μM, 2.54 s-1,
2.92 × 104 M-1s-1, respectively. The purified
laccase was finally used for the selective biotransformation of aromatic methyl
group to aldehyde group in presence of diammonium salt of ABTS as the mediator
and products were characterized by HPLC, IR and
1H NMR. The percentage yields of these transformed products were
>91%.
 
Date 2015-03-30T05:10:17Z
2015-03-30T05:10:17Z
2015-02
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/31263
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.52(1) [February 2015]