来自Cyanothece sp. 的一种新型盐和有机溶剂耐受性酸盐脱酶. 在ATCC 511422中使用
Gamal Nasser Abdel-Hady1,2, Takahisa Tajima1,3, Takeshi Ikeda1
1Unit of Biotechnology, Division of Biological and Life Sciences, Graduate School of Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan.
Frontiers in bioengineering and biotechnology
|September 4, 2023
概括
海洋蓝菌酸盐脱酶 (Ct-PtxD) 提供了增强的NAD(P) H再生. 这种酶表现出优越的盐和有机溶剂耐受性,使其非常适合工业应用.
科学领域:
- 生物化学 生物化学
- 酶学 是一种酶学.
- 生物技术是生物技术.
背景情况:
- 酸盐脱酶 (PtxD) 对于NAD的再生至关重要.
- 扩展PtxD应用需要具有更好的稳定性和耐工业条件的酶.
研究的目的:
- 来克隆,表达和表征来自海洋蓝菌的PtxDCyanothece sp. 在 ATCC 51142 (Ct-PtxD) 里.
- 为了评估Ct-PtxD的稳定性,辅因子偏好和对盐和有机溶剂的耐受性.
- 在具有挑战性的条件下评估Ct-PtxD在合酶反应中的实用性.
主要方法:
- 基因克隆和Ct-PtxD的表达.
- 酶活性测定在各种pH值,温度,盐和有机溶剂度.
- 分子静电潜力和氨基酸组成分析.
- 在结合性性转换试验中证明NADH再生.
主要成果:
- 在pH9.0和50°C时,CT-PtxD表现出最佳活性,具有广泛的pH稳定性 (6.0-10.0).
- 与其他PtxD相比,CT-PtxD表现出对Na+,K+和NH的增强抵抗力.
- 有机溶剂 (乙醇,DMF,甲醇) 增强Ct-PtxD活性,并且耐受性良好.
- 结构分析表明Ct-PtxD的独特特征有助于其稳定性和溶剂耐受性.
- 在高氨条件下,基于Ct-PtxD的NADH再生对合合成有效.
结论:
- Ct-PtxD是一种强大的酶,具有显著的NAD(P) H再生潜力.
- 它对盐和有机溶剂的耐受性使其适合于工业生物转化.
- 在苛刻的反应环境中,CT-PtxD比现有的PtxD具有优势.
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