Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul Technical University, Istanbul, Turkey.
Prep Biochem Biotechnol. 2012;42(6):507-19. doi: 10.1080/10826068.2012.659102.
NAD⁺-dependent formate dehydrogenase (FDH, EC 1.2.1.2) is of use in the regeneration of NAD(P)H coenzymes, and therefore has strong potential for practical application in chemical and medical industries. A low-cost production of recombinant Escherichia coli (E. coli) containing FDH from Candida methylica (cmFDH) was optimized in molasses-based medium by using response surface methodology (RSM) based on central composite design (CCD). The beet molasses as a sole carbon source, (NH₄)₂HPO₄ as a nitrogen and phosphorus source, KH₂PO₄ as a buffer agent, and Mg₂SO₄ · 7H₂O as a magnesium and sulfur source were used as variables in the medium. The optimum medium composition was found to be 34.694 g L⁻¹ of reducing sugar (equivalent to molasses solution), 8.536 g L⁻¹ of (NH₄)₂HPO₄, 3.073 g L⁻¹ of KH₂PO₄, and 1.707 g L⁻¹ of Mg₂SO₄ · 7H₂O. Molasses-based culture medium increased the yield of cmFDH about three times compared to LB medium. The currently developed media has the potential to be used in industrial bioprocesses with low-cost production.
NAD⁺依赖性甲酸盐脱氢酶（FDH，EC 1.2.1.2）在 NAD(P)H 辅酶的再生中具有重要作用，因此在化学和医疗行业具有很强的实际应用潜力。通过基于中心组合设计（CCD）的响应面法（RSM），在以甜菜糖蜜为基础的培养基中对含有来源于 Candida methylica 的 FDH（cmFDH）的重组大肠杆菌（E. coli）进行了优化。将甜菜糖蜜作为唯一的碳源，（NH₄）₂HPO₄作为氮源和磷源，KH₂PO₄作为缓冲剂，Mg₂SO₄·7H₂O 作为镁源和硫源作为培养基中的变量。发现最佳培养基组成为 34.694 g/L 的还原糖（相当于糖蜜溶液）、8.536 g/L 的（NH₄）₂HPO₄、3.073 g/L 的 KH₂PO₄和 1.707 g/L 的 Mg₂SO₄·7H₂O。与 LB 培养基相比，基于糖蜜的培养基使 cmFDH 的产量增加了约三倍。目前开发的培养基具有在低成本生产的工业生物工艺中使用的潜力。
