Abstract Methyl (R)-3-hydroxytetradeconoate ((R)-MHOT) is a crucial chiral intermediate for the chemical synthesis of the anti-obesity drug, orlistat. Here, (R)-MHOT was prepared from methyl 3-oxotetradecanoate (MOT) using a mutant of the short-chain dehydrogenase/reductase (SDR) from Novosphingobium aromaticivorans (NaSDR). Mutant NaSDR-G145A/I199L had a 3.23 times greater kcat value than that of wild type toward MOT. The conditions for the expression of recombinant NaSDR-G145A/I199L were further investigated and obtained cells were used for gram-scale preparation of (R)-MHOT with 50 g/L of MOT. The target product was extracted and confirmed by gas chromatography; the enantiomeric excess value of (R)-MHOT was 99.0 %. Molecular docking analysis was used to reveal the molecular basis of the enhanced catalytic activity of NaSDR-G145A/I199L; NaSDR-G145A/I199L presented a more effective docking posture than NaSDR. This is the first reported use of SDR for preparing (R)-MHOT via the reduction of MOT. Our study provides a foundation for greener preparation of (R)-MHOT.

中文翻译：

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使用来自新鞘氨醇的突变短链脱氢酶酶促合成奥利司他中间体

摘要 (R)-3-羟基十四酸甲酯 ((R)-MHOT) 是化学合成抗肥胖药物奥利司他的关键手性中间体。在这里，(R)-MHOT 是使用来自新鞘氨醇 (NaSDR) 的短链脱氢酶/还原酶 (SDR) 的突变体从 3-氧代十四酸甲酯 (MOT) 制备的。突变体 NaSDR-G145A/I199L 对 MOT 的 kcat 值是野生型的 3.23 倍。进一步研究了重组NaSDR-G145A/I199L的表达条件，并将获得的细胞用于具有50g/L MOT的(R)-MHOT的克级制备。目的产物经气相色谱提取确认；(R)-MHOT的对映体过量值为99.0%。分子对接分析揭示了NaSDR-G145A/I199L催化活性增强的分子基础；NaSDR-G145A/I199L 呈现出比 NaSDR 更有效的对接姿势。这是首次报道使用 SDR 通过减少 MOT 来制备 (R)-MHOT。我们的研究为更环保地制备 (R)-MHOT 提供了基础。