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    • 7. 发明申请
    • THERMOSTABLE NUCLEASE
    • 热稳定核
    • WO2014131113A1
    • 2014-09-04
    • PCT/CA2014/000156
    • 2014-02-27
    • UNIVERSITE LAVAL
    • BOISSINOT, MauriceBOISSINOT, KarelISABEL, SandraSHI, Lu-E
    • C12N15/55A23L1/229A61K31/7088C12N9/22C12P19/40
    • C12N9/22A23L27/23A61K31/7088C12P19/32C12P19/34C12Y301/04
    • The present invention provides a heat-stable nuclease found in Y. enterocolitica subsp. palearctica that is active in broad spectrum conditions. Using the genome sequences of Y. enterocolitica subsp. enterocolitica , we identified one nuclease named Nucyep . This gene was sequenced in a strain Y. enterocolitica subsp. palearctica , cloned, and expressed in E. coli , and then purified and characterized. The molecular weight of this enzyme is about 30 to 32 kDa. The translation product, a nuclease referred to as "Nucyep" is biologically active. The purified Nucyep1 exhibits non-specific nuclease activity, being able to degrade various nucleic acids, including RNA, single-stranded DNA (ssDNA) and linear or circular double-stranded DNA (dsDNA). This enzyme is active in a wide range of temperatures, from 0 to 100°C. The enzyme is active in a wide range of pH values from 3.6 to 9.9, and keeps greater than 75% of the activity at pH 7.24. This enterobacterial nuclease has unique levels of intrinsic resistance to heat, and is active under a large spectrum of conditions.
    • 本发明提供了一种在小肠结肠炎小肠亚种中发现的热稳定核酸酶。 在广谱条件下活跃的棕榈属。 使用小肠结肠炎亚种的基因组序列 我们确定了一种名叫Nucyep的核酸酶。 该基因在小肠结肠炎菌株亚种中测序。 在大肠杆菌中克隆并表达,然后纯化和表征。 该酶的分子量为约30至32kDa。 翻译产品,称为“Nucyep”的核酸酶具有生物活性。 纯化的Nucyep1显示非特异性核酸酶活性,能够降解各种核酸,包括RNA,单链DNA(ssDNA)和线性或环状双链DNA(dsDNA)。 该酶在0至100℃的宽范围的温度范围内是有活性的。 该酶在宽范围的3.6至9.9的pH范围内是有活性的,并且在pH 7.24下保持大于活性的75%。 这种肠杆菌核酸酶具有独特的固有抗热性,并且在大范围的条件下是有活性的。