title: Improved Candida methylica formate dehydrogenase fermentation through statistical optimization of low-cost culture media.
abstract: 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.
authors.affiliation: Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul Technical University, Istanbul, Turkey.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Kahraman
authors.forename: Emel Bıçakçı
authors.initials: EB
authors.lastname: Ordu
authors.forename: Koray
authors.initials: K
authors.lastname: Yeşiladalı
authors.forename: Nevin Gül
authors.initials: NG
authors.lastname: Karagüler
authors.forename: Candan
authors.initials: C
authors.lastname: Tamerler
mesh_terms: D027461:Beta vulgaris / Q000378:metabolism; D002021:Buffers; D002175:Candida / Q000201:enzymology* / Q000378:metabolism; D002244:Carbon / Q000378:metabolism; D003470:Culture Media / Q000378:metabolism / Q000592:standards*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D005560:Formate Dehydrogenases / Q000235:genetics / Q000378:metabolism*; D016015:Logistic Models; D008278:Magnesium Sulfate / Q000378:metabolism; D008965:Molasses / Q000032:analysis; D009584:Nitrogen / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D017680:Potassium Compounds / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D002021:Buffers; D003470:Culture Media; D010710:Phosphates; D017680:Potassium Compounds; D011994:Recombinant Proteins; C024788:ammonium phosphate; D002244:Carbon; D008278:Magnesium Sulfate; C013216:potassium phosphate; D005560:Formate Dehydrogenases; D009584:Nitrogen