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Production, Gene Cloning, and Overexpression of a Laccase in the Marine-Derived Yeast Aureobasidium melanogenum Strain 11-1 and Characterization of the Recombinant Laccase

  • Thu Aung
  • Hong Jiang
  • Cheng-Cheng Chen
  • Guang-Lei Liu
  • Zhong Hu
  • Zhen-Ming ChiEmail author
  • Zhe ChiEmail author
Original Article
  • 129 Downloads

Abstract

Aureobasidium melanogenum strain 11-1 with a high laccase activity was isolated from a mangrove ecosystem. Under the optimal conditions, the 11-1 strain yielded the highest laccase activity up to 3120.0 ± 170 mU/ml (1.2 U/mg protein) within 5 days. A laccase gene (LAC1) of the yeast strain 11-1 contained two introns and encoded a protein with 570 amino acids and four conserved copper-binding domains typical of the fungal laccase. Expression of the LAC1 gene in the yeast strain 11-1 made a recombinant yeast strain produce the laccase activity of 6005 ± 140 mU/ml. The molecular weight of the recombinant laccase after removing the sugar was about 62.5 kDa. The optimal temperature and pH of the recombinant laccase were 40 °C and 3.2, respectively, and it was stable at a temperature less than 25 °C. The laccase was inhibited in the presence of sodium dodecyl sulfate (SDS), ethylenediaminetetraacetic acid (EDTA), phenylmethanesulfonyl fluoride (PMSF), and dl-dithiothreitol (DTT). The Km and Vmax values of the laccase for 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was 6.3 × 10−2 mM and 177.4 M/min, respectively. Many synthetic dyes were greatly decolored by the laccase.

Keywords

Laccase Aureobasidium melanogenum LAC1 gene Overexpression Decolorizing ability 

Notes

Funding Information

This study was financially supported by the National Natural Science Foundation of China (Grant No. 31561163001).

Supplementary material

10126_2018_9860_MOESM1_ESM.doc (384 kb)
ESM 1 (DOC 384 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Key Laboratory of Marine Genetics and Breeding (Ocean University of China)Ministry of EducationQingdaoChina
  2. 2.Laboratory for Marine Biology and BiotechnologyQingdao National Laboratory for Marine Science and TechnologyQingdaoChina
  3. 3.Department of BiologyShantou UniversityShantouChina

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