Identification of l-histidine oxidase activity in Achromobacter sp. TPU 5009 for l-histidine determination.

An enzyme showing l-histidine oxidase (HisO) activity by the formation of hydrogen peroxide was newly purified from Achromobacter sp. TPU 5009. This enzyme was found to be a heterodimer of two proteins (molecular mass, 53.8 and 58.3 kDa), the partial determination of which indicated they are homologs of l-histidine ammonia-lyase (AchHAL) and urocanate hydratase (AchURO). The enzyme was stable in a pH range of 5.0-11.0, with >90% of the original activity maintained below 60°C at pH 7.0. To characterize AchHAL and AchURO, each of their genes was cloned and expressed in a heterologous expression system. Heterologous AchHAL catalyzed the elimination of the α-amino group of l-histidine to urocanate and ammonia, while heterologous AchURO catalyzed the hydration of urocanate to imidazolone propionate. Since imidazolone propionate is highly unstable in the presence of oxygen at neutral pH, it was immediately decomposed and hydrogen peroxide was non-enzymatically produced. Our results indicate that this natural enzyme showing apparent HisO activity is composed of AchHAL and AchURO, which formed hydrogen peroxide after the spontaneous decomposition of imidazolone propionate.
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