title: Heterologous expression and characterisation of a laccase from Colletotrichum lagenarium and decolourisation of different synthetic dyes.
abstract: Laccases have received considerable attention in recent decades because of their ability to oxidise a large spectrum of phenolic and non-phenolic organic substrates and highly recalcitrant environmental pollutants. In this research, a laccase gene from Colletotrichum lagenarium was chemically synthesised using yeast bias codons and expressed in Pichia pastoris. The molecular mass of the recombinant laccase was estimated to be 64.6 kDa by SDS-PAGE, and the enzyme exhibited maximum activity at pH 3.6-4.0 but more stability in buffer with higher pH (>pH 3.6). The optimal reaction temperature of the enzyme was 40 °C, beyond which stability significantly decreased. By using 2,2'-azino-bis-(3-ethylbenzothiazoline)-6-sulphonate (ABTS) as a substrate, K m and V max values of 0.34 mM and 7.11 mM min(-1) mg(-1), respectively, were obtained. Using ABTS as a mediator, the laccase could oxidise hydroquinone to p-benzoquinone and decolourise the synthetic dyes malachite green, crystal violet and orange G. These results indicated that the laccase could be used to treat industrial effluents containing artificial dyes.
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China. wangbosaas@163.com.
authors: Bo
authors: B
authors: Wang
authors: Crop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201403, China.
authors: Ying
authors: Y
authors: Yan
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.
authors: Yongsheng
authors: Y
authors: Tian
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.
authors: Wei
authors: W
authors: Zhao
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.
authors: Zhengjun
authors: Z
authors: Li
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.
authors: Jianjie
authors: J
authors: Gao
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China.
authors: Rihe
authors: R
authors: Peng
authors: Biotechnological Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, China. yao.quanhong65@yahoo.com.
authors: Quanhong
authors: Q
authors: Yao
mesh_terms: D052160:Benzothiazoles / Q000378:metabolism; D001711:Biotransformation; D020231:Colletotrichum / Q000201:enzymology* / Q000235:genetics; D004396:Coloring Agents / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D042845:Laccase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008970:Molecular Weight; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D004718:Saccharomycetales; D013379:Substrate Specificity; D013451:Sulfonic Acids / Q000378:metabolism; D013696:Temperature
chemical_list: D052160:Benzothiazoles; D004396:Coloring Agents; D011994:Recombinant Proteins; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D042845:Laccase
keywords: Characterisation; Colletotrichum lagenarium; Laccase; Pichia pastoris; Synthetic dyes