title: A highly efficient recombinant laccase from the yeast Yarrowia lipolytica and its application in the hydrolysis of biomass.
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.
authors.affiliation: Department of Chemical Engineering, Konkuk University, Seoul, Korea.
authors.forename: Dayanand
authors.initials: D
authors.lastname: Kalyani
authors.affiliation: Department of Chemical Engineering, Konkuk University, Seoul, Korea.
authors.forename: Manish Kumar
authors.initials: MK
authors.lastname: Tiwari
authors.affiliation: Department of Chemical Engineering, Konkuk University, Seoul, Korea.
authors.forename: Jinglin
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yuseong-gu, Daejeon, Korea.
authors.forename: Sun Chang
authors.initials: SC
authors.lastname: Kim
authors.affiliation: Microbial Biotechnology and Genomics, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi, India.
authors.forename: Vipin C
authors.initials: VC
authors.lastname: Kalia
authors.affiliation: Department of Materials Science and Engineering, Korea University, Anam-Dong, Seongbuk-Gu, Seoul, Korea.
authors.forename: Yun Chan
authors.initials: YC
authors.lastname: Kang
authors.affiliation: Department of Chemical Engineering, Konkuk University, Seoul, Korea.
authors.forename: Jung-Kul
authors.initials: JK
authors.lastname: Lee
mesh_terms: D018533:Biomass; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D006868:Hydrolysis; D042845:Laccase / Q000737:chemistry* / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D032107:Populus / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014934:Wood / Q000737:chemistry*; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D042845:Laccase