Istituto di Biochimica delle Proteine, CNR, Via P. Castellino 111, 80131 Naples, Italy.
Appl Biochem Biotechnol. 2013 Jul;170(6):1482-90. doi: 10.1007/s12010-013-0282-3. Epub 2013 May 19.
The synthesis of the aroma chemical cinnamyl alcohol (CMO) by means of enzymatic reduction of cinnamaldehyde (CMA) was investigated using NADH-dependent alcohol dehydrogenase from Bacillus stearothermophilus both as an isolated enzyme, and in recombinant Escherichia coli whole cells. The influence of parameters such as reaction time and cofactor, substrate, co-substrate 2-propanol and biocatalyst concentrations on the bioreduction reaction was investigated and an efficient and sustainable one-phase system developed. The reduction of CMA (0.5 g/L, 3.8 mmol/L) by the isolated enzyme occurred in 3 h at 50 °C with 97% conversion, and yielded high purity CMO (≥98%) with a yield of 88% and a productivity of 50 g/genzyme. The reduction of 12.5 g/L (94 mmol/L) CMA by whole cells in 6 h, at 37 °C and no requirement of external cofactor occurred with 97% conversion, 82% yield of 98% pure alcohol and a productivity of 34 mg/gwet cell weight. The results demonstrate the microbial system as a practical and efficient method for larger-scale synthesis of CMO.
采用嗜热脂肪芽孢杆菌 NADH 依赖型醇脱氢酶(Bacillus stearothermophilus NADH-dependent alcohol dehydrogenase),通过酶法还原肉桂醛(cinnamaldehyde,CMA)合成芳樟醇(cinnamyl alcohol,CMO),分别考察了游离酶和重组大肠杆菌全细胞作为生物催化剂时,反应时间、辅酶、底物、共底物 2-丙醇以及生物催化剂浓度等参数对生物还原反应的影响,开发了一种高效、可持续的单相体系。在 50℃下,游离酶反应 3 h 可使肉桂醛(0.5 g/L,3.8 mmol/L)完全转化,产物芳樟醇(≥98%)的纯度为 88%,酶的得率为 88%,酶的生产效率为 50 g/genzyme。重组大肠杆菌全细胞在 37℃下反应 6 h,无需外加辅酶,肉桂醛(12.5 g/L,94 mmol/L)转化率可达 97%,产物芳樟醇(98%纯度)的得率为 82%,酶的生产效率为 34 mg/gwet cell weight。结果表明,该微生物体系是一种用于大规模合成 CMO 的实用、高效的方法。