caa a22 4500 605503745 CHVBK 20210128100612.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-6203-8 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6203-8 Isolation, sequencing, and heterologous expression of the Paecilomyces variotii gene encoding S -hydroxymethylglutathione dehydrogenase ( fldA ) [Elektronische Daten] [Takuji Oka, Yuji Komachi, Kazufumi Ohshima, Yoichi Kawano, Kohsai Fukuda, Kazuhiro Nagahama, Keisuke Ekino, Yoshiyuki Nomura] The filamentous fungus Paecilomyces variotii NBRC 109023 (teleomorph: Byssochlamys spectabilis NBRC 109023) degrades formaldehyde at concentrations as high as 2.4% (w/v). In many prokaryotes and in all known eukaryotes, formaldehyde degradation is catalyzed by S-hydroxymethylglutathione (S-HMGSH) dehydrogenase. We report here the isolation and characterization of the gene encoding S-HMGSH dehydrogenase activity in P. variotii. The 1.6-kb fldA gene contained 5 introns and 6 exons, and the corresponding cDNA was 1143bp, encoding a 40-kDa protein composed of 380 amino acids. FldA was predicted to have 74.3, 73.7, 68.5, and 67.4% amino acid identity to the S-HMGSH dehydrogenases of Hansenula polymorpha, Candida boidinii, Saccharomyces cerevisiae, and Kluyveromyces lactis, respectively. The predicted protein also showed high amino acid similarity (84∼86%) to the products of putative fldA genes from other filamentous fungi, including Aspergillus sp. and Penicillium sp. Notably, the P. variotii fldA gene was able to functionally complement a Saccharomyces cerevisiae strain (BY4741 ∆sfa1) lacking the gene for S-HMGSH dehydrogenase. The heterologous expression construct rendered BY4741 ∆sfa1 tolerant to exogenous formaldehyde. Although BY4741 (parental wild-type strain) was unable to degrade even low concentrations of formaldehyde, BY4741 ∆sfa1 harboring Paecilomyces fldA was able to degrade 4mM formaldehyde within 30h. The findings from this study confirm the essential role of S-HMGSH dehydrogenase in detoxifying formaldehyde. The Author(s), 2014 Paecilomyces variotii nationallicence Formaldehyde degradation nationallicence S -hydroxymethylglutathione dehydrogenase nationallicence fldA nationallicence Heterologous expression nationallicence Oka Takuji Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Komachi Yuji Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Ohshima Kazufumi Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Kawano Yoichi Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Fukuda Kohsai Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Nagahama Kazuhiro Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Ekino Keisuke Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Nomura Yoshiyuki Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1755-1763 0175-7598 99:4<1755 2015 99 253 https://doi.org/10.1007/s00253-014-6203-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6203-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Oka Takuji Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Komachi Yuji Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Ohshima Kazufumi Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Kawano Yoichi Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Fukuda Kohsai Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Nagahama Kazuhiro Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Ekino Keisuke Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 700 1- Nomura Yoshiyuki Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, 860-0082, Kumamoto, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1755-1763 0175-7598 99:4<1755 2015 99 253