title: Molecular breeding of lignin-degrading brown-rot fungus Gloeophyllum trabeum by homologous expression of laccase gene.
abstract: The basidiomycete Gloeophyllum trabeum KU-41 can degrade Japanese cedar wood efficiently. To construct a strain better suited for biofuel production from Japanese cedar wood, we developed a gene transformation system for G. trabeum KU-41 using the hygromycin phosphotransferase-encoding gene (hpt) as a marker. The endogenous laccase candidate gene (Gtlcc3) was fused with the promoter of the G. trabeum glyceraldehyde-3-phosphate dehydrogenase-encoding gene and co-transformed with the hpt-bearing pAH marker plasmid. We obtained 44 co-transformants, and identified co-transformant L#61, which showed the highest laccase activity among all the transformants. Moreover, strain L#61 was able to degrade lignin in Japanese cedar wood-containing medium, in contrast to wild-type G. trabeum KU-41 and to a typical white-rot fungus Phanerochaete chrysosporium. By using strain L#61, direct ethanol production from Japanese cedar wood was improved compared to wild type. To our knowledge, this study is the first report of the molecular breeding of lignin-degrading brown-rot fungus and direct ethanol production from softwoods by co-transformation with laccase overproduction constructs.
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan. misa_ch_2_29@yahoo.co.jp.
authors.forename: Misa
authors.initials: M
authors.lastname: Arimoto
authors.affiliation: NARO National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki, 305-8642, Japan. yamagisi@affrc.go.jp.
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamagishi
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan. o_wangjianqiao@yahoo.co.jp.
authors.forename: Jianqiao
authors.initials: J
authors.lastname: Wang
authors.affiliation: Integrative Technology Research Institute, Teijin Limited, Iwakuni, 740-8511, Japan. kan.tanaka@teijin.co.jp.
authors.forename: Kanade
authors.initials: K
authors.lastname: Tanaka
authors.affiliation: New Business Development Business Unit, Teijin Limited, Tokyo, 100-8585, Japan. ta.miyoshi@teijin.co.jp.
authors.forename: Takanori
authors.initials: T
authors.lastname: Miyoshi
authors.affiliation: Faculty of Agriculture, University of Miyazaki, Miyazaki, 889-2192, Japan. kamei@cc.miyazaki-u.ac.jp.
authors.forename: Ichiro
authors.initials: I
authors.lastname: Kamei
authors.affiliation: Faculty of Agriculture, Kyushu University, Fukuoka, 812-8581, Japan. ryukondo@agr.kyushu-u.ac.jp.
authors.forename: Ryuichiro
authors.initials: R
authors.lastname: Kondo
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan. mori.toshio@shizuoka.ac.jp.
authors.forename: Toshio
authors.initials: T
authors.lastname: Mori
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan. kawagishi.hirokazu@shizuoka.ac.jp.|Graduate School of Science and Technology, Shizuoka University, Shizuoka, 422-8529, Japan. kawagishi.hirokazu@shizuoka.ac.jp.|Research Institute of Green Science and Technology, Shizuoka University, Shizuoka, 422-8529, Japan. kawagishi.hirokazu@shizuoka.ac.jp.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kawagishi
authors.affiliation: Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan. hirai.hirofumi@shizuoka.ac.jp.|Research Institute of Green Science and Technology, Shizuoka University, Shizuoka, 422-8529, Japan. hirai.hirofumi@shizuoka.ac.jp.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hirai
keywords: Gloeophyllum trabeum KU-41; Homologous expression; Laccase gene; Lignin degradation