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嗜热微生物中获取耐乙醛变性醛缩酶 被引量：2

Prospecting for Acetaldehyde-tolerant DERAs from Thermophilic Microorganism
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摘要 在催化他汀药物中间体合成中,对高浓度乙醛底物的耐受是醛缩酶的重要应用性质.通过克隆并在大肠杆菌中过表达了来源于嗜热微生物Geobacillusthermodenitrificans的耐热醛缩酶(DERAGth),该蛋白质序列长度为223aa,对应的分子量为25.0kD.初步酶学性质表征结果表明,该酶对天然底物2-脱氧核酸-5-磷酸的活力19.3U/mg,最适pH为7.5,最适温度为60℃.在40—100℃保温10min的结果表明,DERAGth具有较好的热稳定性,在60℃下10min活力仍保持100%.对乙醛耐受的实验表明,该酶在300mmol乙醛中25℃保温2h,剩余活力高于40%.连续醛缩活力对比实验结果表明,该酶的连续醛缩活力远大于来源于常温微生物的大肠杆菌醛缩酶. Resistance to high concentration acetaldehyde is one of the most important properties for 2-deoxy-D-ribose-5- phosphate aldolase （DERA） in statin synthesis. The themophilic DERA gene from Geobacillus thermodenitrificans is subcloned and overexpressed in E. coli. The sequence of this gene encoded 223aa amino acids predicts to yield protein of molecular mass 25. 0 kDa. The specific activity of DERAGth is 19. 3 U/rag for its nature substrate 2-deoxyribose-5- phosphate substrate. DERAGth activity is optimal at pH 7.5 and 60 ℃. The thermophilic enzyme is heat stable no activity loss is observed after incubation at 60 ℃ for 10 min. The thermophilic enzyme also has showed a high resistance to acetaldehyde retained more than 40% activity after exposure for 2 h to 300 mmol acetaldehyde at 25 ℃. In compared analysis, DERAGth shows significantly higher aldol condensation activity than that of DERA from E. coll.
作者 李铖璐 杜鹏飞 裴晓林 吴慧丽 谢开林 王秋岩
机构地区 杭州师范大学生物医药与健康研究中心
出处 《杭州师范大学学报（自然科学版）》 CAS 2010年第5期372-378,共7页 Journal of Hangzhou Normal University(Natural Science Edition)
关键词 2-脱氧-D-核酸-5-磷酸醛缩酶 连续醛缩 嗜热酶 乙醛抗性 2-deoxy-D-ribose 5-phosphate aldolase sequential aldol condensation themophilic enzyme acetaldehyde toleranc
分类号 Q819 [生物学—生物工程]
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参考文献21

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同被引文献25

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杭州师范大学学报（自然科学版）
2010年 第5期

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