Biochemical characterization of a catalase from Antrodia camphorata: Expression in Escherichia coli and enzyme properties

Chuian-Fu Ken, Hsueh Tai Chen, Reny Chang Chang, Chi Tsai Lin

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Catalase plays important roles in antioxidation and cell signaling. One cDNA (1794 bp, DQ021914) encoding the putative catalase was cloned from Antrodia camphorata. The deduced amino acid sequence is conserved among the reported catalases. To characterize the A. camphorata catalase, the coding region was subcloned into a vector pET-20b(+) and transformed into E. coli. The recombinant 6His-tagged catalase was expressed and purified by Ni 2+ - nitrilotriacetic acid sepharose. The purified enzyme showed one band by 10% SDS-PAGE. The enzyme retained 50% activity at 60°C for 14 min. The enzyme was active under a broad pH range from 7.8 to 11.2. The enzyme showed 67% activity after 4 h of incubation at 37°C with trypsin. It was also proven able to protect intact supercoiled plasmid DNA from ·OH-induced nicking. Study of the enzyme's properties may prove beneficial for future applications in medicine or health food.

Original languageEnglish
Pages (from-to)119-125
Number of pages7
JournalBotanical Studies
Volume49
Issue number2
Publication statusPublished - 2008 Apr 1

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Taiwanofungus camphoratus
catalase
Escherichia coli
enzymes
nitrilotriacetic acid
health foods
trypsin
agarose
plasmids
medicine
amino acid sequences

All Science Journal Classification (ASJC) codes

  • Plant Science

Cite this

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abstract = "Catalase plays important roles in antioxidation and cell signaling. One cDNA (1794 bp, DQ021914) encoding the putative catalase was cloned from Antrodia camphorata. The deduced amino acid sequence is conserved among the reported catalases. To characterize the A. camphorata catalase, the coding region was subcloned into a vector pET-20b(+) and transformed into E. coli. The recombinant 6His-tagged catalase was expressed and purified by Ni 2+ - nitrilotriacetic acid sepharose. The purified enzyme showed one band by 10{\%} SDS-PAGE. The enzyme retained 50{\%} activity at 60°C for 14 min. The enzyme was active under a broad pH range from 7.8 to 11.2. The enzyme showed 67{\%} activity after 4 h of incubation at 37°C with trypsin. It was also proven able to protect intact supercoiled plasmid DNA from ·OH-induced nicking. Study of the enzyme's properties may prove beneficial for future applications in medicine or health food.",
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Biochemical characterization of a catalase from Antrodia camphorata : Expression in Escherichia coli and enzyme properties. / Ken, Chuian-Fu; Chen, Hsueh Tai; Chang, Reny Chang; Lin, Chi Tsai.

In: Botanical Studies, Vol. 49, No. 2, 01.04.2008, p. 119-125.

Research output: Contribution to journalArticle

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