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A bifunctional enzyme of gluthathione‐dependent formaldehyde dehydrogenase and S‐nitrosogluthathione reductase from Antrodia camphorata
Author(s) -
Wen Lisa,
Huang ChihYu,
Ken ChuianFu,
Lin ChiTsai
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.853.2
Subject(s) - formaldehyde dehydrogenase , enzyme , biochemistry , nitrilotriacetic acid , reductase , microbiology and biotechnology , chemistry , glutathione , bifunctional , biology , chelation , organic chemistry , catalysis
Glutathione‐dependent formaldehyde dehydrogenase (GFD) plays important roles in formaldehyde detoxification and antioxidation. A gene encoding GFD from A. camphorate was identified based on sequence homology. The deduced amino acid sequence of 378 residues is conserved among the reported GFDs. To characterize the Ac‐GFD, the coding region was subcloned into a vector pET‐20b(+) and transformed into E. coli. The recombinant GFD was expressed and purified by Ni 2+ ‐nitrilotriacetic acid Sepharose. This purified enzyme showed a single band on a 10 % SDS‐PAGE. The enzyme retained 50% GFD activity after heating at 50 o C for 5 min. The enzyme is bifunctional. In addition to the GFD activity, it also functions as an effective S‐nitrosoglutathione reductase (GSNOR) presumably to safeguard against nitrosative stress. The Km values for S‐hydroxymethylglutathione and S‐ nitrosoglutathione were 1.197 mM and 0.276 mM, respectively. Supported by the National Science Council of the Republic of China under grant NSC 96‐2313‐B‐019‐001 to C‐T. L.