A highly efficient recombinant laccase from the yeast Yarrowia lipolytica and its application in the hydrolysis of biomass

Dayanand Kalyani; Manish Kumar Tiwari; Jinglin Li; Sun Chang Kim; Vipin C Kalia; Yun Chan Kang; Jung-Kul Lee

doi:10.1371/journal.pone.0120156

A highly efficient recombinant laccase from the yeast Yarrowia lipolytica and its application in the hydrolysis of biomass

PLoS One. 2015 Mar 17;10(3):e0120156. doi: 10.1371/journal.pone.0120156. eCollection 2015.

Authors

Dayanand Kalyani¹, Manish Kumar Tiwari¹, Jinglin Li¹, Sun Chang Kim², Vipin C Kalia³, Yun Chan Kang⁴, Jung-Kul Lee¹

Affiliations

¹ Department of Chemical Engineering, Konkuk University, Seoul, Korea.
² Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yuseong-gu, Daejeon, Korea.
³ Microbial Biotechnology and Genomics, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi, India.
⁴ Department of Materials Science and Engineering, Korea University, Anam-Dong, Seongbuk-Gu, Seoul, Korea.

Abstract

A modified thermal asymmetric interlaced polymerase chain reaction was performed to obtain the first yeast laccase gene (YlLac) from the isolated yeast Yarrowia lipolytica. The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris. YlLac is a monomeric glycoprotein with a molecular mass of ~55 kDa as determined by polyacrylamide-gel electrophoresis. It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s(-1) μM(-1)) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s(-1) μM(-1)) than other reported laccases. The standard redox potential of the T1 site of the enzyme was found to be 772 mV. The highest catalytic efficiency of the yeast recombinant laccase, YlLac, makes it a good candidate for industrial applications: it removes phenolic compounds in acid-pretreated woody biomass (Populus balsamifera) and enhanced saccharification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biomass*
Fungal Proteins / chemistry*
Fungal Proteins / genetics
Hydrolysis
Laccase / chemistry*
Laccase / genetics
Pichia / genetics
Pichia / metabolism
Populus / chemistry*
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Wood / chemistry*
Yarrowia / enzymology*
Yarrowia / genetics

Substances

Fungal Proteins
Recombinant Proteins
Laccase

Grants and funding

This work was supported by the Energy Efficiency & Resources Core Technology Program of the Korea Institute of Energy Technology Evaluation and Planning (KETEP), granted financial resource from the Ministry of Trade, Industry & Energy, Republic of Korea (201320200000420). This research was also supported by a grant from the Intelligent Synthetic Biology Center of Global Frontier Project (2011-0031955) funded by the Ministry of Science, ICT and Future Planning, Republic of Korea. This work was supported by 2013 KU Brain Pool fellowship of Konkuk University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.