脱氧核糖醛缩酶与醇脱氢酶共催化合成(3R,5S)-6-氯-2,4,6-三脱氧己内酯

Co-Catalysis of Deoxyribose Aldolaseand Alcohol Dehydrogenase to Generate (3R,5S)-6-Chloro-2,4,6-Trideoxy-Erythro-Hexonolactone

摘要:

为实现生物法“一步”合成他汀类药物侧链中间体（3R，5S）-6-氯-2，4，6-三脱氧己内酯，采用生物偶联的方法分别构建不同抗性的两种重组质粒，即来源于Streptococcus suis的脱氧核糖醛缩酶基因与Lodderomyces elongisporus的醇脱氢酶基因的重组质粒，将两种重组质粒共转化于大肠杆菌 BL21同一细胞中，实现两种酶的可溶性共表达。以重组大肠杆菌为生物催化剂，全细胞催化两种底物：400 mmol/L乙醛与200 mmol/L氯乙醛发生反应，获得产物（3R，5S）-6-氯-2，4，6-三脱氧己内酯。在缺少标准产物对照的条件下，利用质谱（MS）以及氢谱（¹H-NMR）进行产物鉴定，最终确定合成的产物为他汀类药物侧链中间体（3R，5S）-6-氯-2，4，6-三脱氧己内酯。

Abstract:

The biological coupling method was used in tandem with the enzyme reaction process to synthesize (3R,5S)-6-chloro-2,4,6-trideoxy-erythro-hexonolactone, the statin side-chain intermediate, in 'one-step' bio-synthesis. The recombinant plasmids of Streptococcus suis deoxyribose aldolase gene and Lodderomyces elongisporus alcohol dehydrogenase gene were co-transformedinto the same cell of Escherichia coli BL21 to realize their soluble co-expression.(3R,5S)-6-chloro-2,4,6-trideoxy-erythro-hexonolactone was obtained by the whole cell co-catalysis of 400 mmol/L acetaldehyde and 200 mmol/L chloroacetaldehyde using recombinant E. coli was used as the biocatalysts. In the absence of standard products, mass spectrometry and 1H nuclear magnetic resonance were performed to identifythe products. The final product was determined as the statin side-chain intermediate (3R,5S)-6-chloro-2,4,6-trideoxy-erythro-hexonolactone.