摘要
试验以来源于嗜热菌DSM 13965的磷酸三酯酶样内酯酶基因在毕赤酵母GS115中进行表达,旨在得到嗜热磷酸三酯酶样内酯酶,经组氨酸标签镍柱法纯化后,从底物非专一性、热稳定性、最适温度及最适pH、pH稳定性等方面进行酶的性质表征。结果表明:嗜热菌磷酸三酯酶样内酯酶具有水解磷酸三酯与内酯的双重水解能力;最适温度为75℃;最适pH为7;在75℃下半衰期为8 h,在碱性条件下的稳定性高于酸性条件。因此嗜热磷酸三酯酶具有较高热稳定性及可溶性,能够在实际的工业应用中抵抗苛刻的工业生产条件,因而具有更好的应用潜力。
In this artical, we gained the phosphotriesterase-like lactonase （PPL） Gene from thermophiles Thermaerobacter subterraneus DSM 13965, then cloned and expressed at high levels in pichia yeast GS115. Purified by his-tag in ni-sepharose chromatography, the activity of recombinant protein was detectived from following aspects： two subtrates; thermostability, temperature and pH, pH stability. The results showed that： it can both hydrolysis phosphate ester and lactone abilityoptimum temperature and pH were 75 ℃ and 7.0, respectively, and the activity exited after putting in the 75 ℃ for 8 h, has a higher alkaline stability. With a high heat and soluble stability, PPL can resist harsh conditions of industrial production in actual industrial application, thus has better application potential.
出处
《食品工业》
北大核心
2014年第8期184-189,共6页
The Food Industry
