title: Laccase Gene Family in Cerrena sp. HYB07: Sequences, Heterologous Expression and Transcriptional Analysis.
abstract: Laccases are a class of multi-copper oxidases with industrial potential. In this study, eight laccases (Lac1-8) from Cerrena sp. strain HYB07, a white-rot fungus with high laccase yields, were analyzed. The laccases showed moderate identities to each other as well as with other fungal laccases and were predicted to have high redox potentials except for Lac6. Selected laccase isozymes were heterologously expressed in the yeast Pichia pastoris, and different enzymatic properties were observed. Transcription of the eight laccase genes was differentially regulated during submerged and solid state fermentation, as shown by quantitative real-time polymerase chain reaction and validated reference genes. During 6-day submerged fermentation, Lac7 and 2 were successively the predominantly expressed laccase gene, accounting for over 95% of all laccase transcripts. Interestingly, accompanying Lac7 downregulation, Lac2 transcription was drastically upregulated on days 3 and 5 to 9958-fold of the level on day 1. Consistent with high mRNA abundance, Lac2 and 7, but not other laccases, were identified in the fermentation broth by LC-MS/MS. In solid state fermentation, less dramatic differences in transcript abundance were observed, and Lac3, 7 and 8 were more highly expressed than other laccase genes. Elucidating the properties and expression profiles of the laccase gene family will facilitate understanding, production and commercialization of the fungal strain and its laccases.
authors.affiliation: Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou University, Fuzhou 350116, Fujian, China. T09136@fzu.edu.cn.
authors.forename: Jie
authors.initials: J
authors.lastname: Yang
authors.affiliation: Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou University, Fuzhou 350116, Fujian, China. xuxinqi@fzu.edu.cn.
authors.forename: Xinqi
authors.initials: X
authors.lastname: Xu
authors.affiliation: School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China. b021770@mailserv.cuhk.edu.hk.
authors.forename: Tzi Bun
authors.initials: TB
authors.lastname: Ng
authors.affiliation: Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou University, Fuzhou 350116, Fujian, China. ljuan@fzu.edu.cn.
authors.forename: Juan
authors.initials: J
authors.lastname: Lin
authors.affiliation: Fujian Key Laboratory of Marine Enzyme Engineering, Fuzhou University, Fuzhou 350116, Fujian, China. xiuyunye@fzu.edu.cn.
authors.forename: Xiuyun
authors.initials: X
authors.lastname: Ye
mesh_terms: D020134:Catalytic Domain; D003001:Cloning, Molecular; D005285:Fermentation; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D042845:Laccase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D005810:Multigene Family; D011124:Polyporaceae / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D005656:Fungal Proteins; D042845:Laccase
keywords: Cerrena sp.; fermentation; heterologous expression; laccase; qPCR