Secretory expression of the non-secretory-type Lentinula edodes laccase by Aspergillus oryzae.
The shiitake mushroom, Lentinula edodes, has an extracelluar secretory-type laccase, Lcc1, and a fruiting-body-accumulation-type laccase, Lcc4. We previously reported the production of Lcc1 by plant cells, but had difficulty producing Lcc4. Here, we report the production of Lcc1 and Lcc4 by Aspergillus oryzae and the extracellular secretory production of Lcc4 using a modified secretion signal peptide (SP) from Lcc1. Sp-Lcc4 produced by A. oryzae had biochemical activities similar to Lcc4 produced by L. edodes. Lcc1 did not react with beta-(3,4-dihydroxyphenol) alanine (DOPA), but Lcc4 from L. edodes and A. oryzae could oxidize DOPA. K(M) values for the substrates 2,2'-azino-di-(3-ethylbenzthiazolinsulfonate), 2,6-dimethoxyphenol, guaiacol, pyrogallol, and catechol were similar for Lcc4 and Sp-Lcc4. In conclusion, a non-secretory-type fungal laccase is secreted into the culture media with its original enzymatic properties by exploiting modified secretory signal peptide.
Yano
Akira
A
Iwate Biotechnology Research Center, 22-174-4 Narita, Kitakami, Iwate 024-0003, Japan. akiray@ibrc.or.jp
Kikuchi
Sayaka
S
Nakagawa
Yuko
Y
Sakamoto
Yuichi
Y
Sato
Toshitsugu
T
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D048690:Fruiting Bodies, Fungal / Q000201:enzymology; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D007700:Kinetics; D042845:Laccase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D015202:Protein Engineering; D021382:Protein Sorting Signals; D021381:Protein Transport; D016415:Sequence Alignment; D020942:Shiitake Mushrooms / Q000201:enzymology*; D013379:Substrate Specificity
D005656:Fungal Proteins; D021382:Protein Sorting Signals; D042845:Laccase