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

PloS One
Dayanand KalyaniJung-Kul Lee

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.

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Citations

Dec 25, 2015·Molecular Biotechnology·Zuzana Antošová, Hana Sychrová
Dec 29, 2015·Trends in Biotechnology·Sergio Rosales-MendozaBeatriz Meza
Oct 29, 2015·Bioprocess and Biosystems Engineering·Marwa M EldalatonyByong-Hun Jeon
Jul 14, 2016·Indian Journal of Microbiology·Radhika DeshmukhHemant J Purohit
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Feb 6, 2017·Indian Journal of Microbiology·Vijaya GuptaPrince Sharma
Jan 23, 2021·Microbial Cell Factories·Michael Dare AsemoloyeLorenzo Pecoraro
Mar 14, 2021·Environmental Science and Pollution Research International·Manisha Malhotra, Sunil Kumar Suman
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Jul 24, 2021·Indian Journal of Microbiology·Gurusamy MuneeswaranIn-Won Kim
Dec 23, 2016·ACS Applied Materials & Interfaces·Sanjay K S PatelJung-Kul Lee

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Datasets Mentioned

BETA
JF719545
ABN13592
ACR50978
AAM18408
BAD98307
AAG09231

Methods Mentioned

BETA
PCR
electrophoresis
chips
gel filtration

Software Mentioned

- 3D Score / Verify Protein
ScanProsite
PROCHECK
Compute pI / Mw
YlLac
BLAST
ClustalW2
DS
GraphPad Prism
Discovery Studio

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