Stark D, Münch T, Sonnleitner B, Marison I W, von Stockar U
Laboratory of Chemical and Biochemical Engineering, Swiss Federal Institute of Technology (EPFL), CH-1015 Lausanne, Switzerland.
Biotechnol Prog. 2002 May-Jun;18(3):514-23. doi: 10.1021/bp020006n.
The bioconversion of L-phenylalanine (L-Phe) to 2-phenylethanol (PEA) by the yeast Saccharomyces cerevisiae is limited by the toxicity of the product. PEA extraction by a separate organic phase in the fermenter is the ideal in situ product recovery (ISPR) technique to enhance productivity. Oleic acid was chosen as organic phase for two-phase fed-batch cultures, although it interfered to some extent with yeast viability. There was a synergistic inhibitory impact toward S. cerevisiae in the presence of PEA, and therefore a maximal PEA concentration in the aqueous phase of only 2.1 g/L was achieved, compared to 3.8 g/L for a normal fed-batch culture. However, the overall PEA concentration in the fermenter was increased to 12.6 g/L, because the PEA concentration in the oleic phase attained a value of 24 g/L. Thus, an average volumetric PEA production rate of 0.26 g L(-1) h(-1) and a maximal volumetric PEA production rate of 0.47 g L(-1) h(-1) were achieved in the two-phase fed-batch culture. As ethanol inhibition had to be avoided, the production rates were limited by the intrinsic oxidative capacity of S. cerevisiae. In addition, the high viscosity of the two-phase system lowered the k(l)a, and therefore also the productivity. Thus, if a specific ISPR technique is planned, it consequently has to be remembered that the productivity of this bioconversion process is also quickly limited by the k(l)a of the fermenter at high cell densities.
酿酒酵母将L-苯丙氨酸(L-Phe)生物转化为2-苯乙醇(PEA)的过程受产物毒性限制。在发酵罐中用单独的有机相萃取PEA是提高生产率的理想原位产物回收(ISPR)技术。油酸被选作两相补料分批培养的有机相,尽管它在一定程度上会影响酵母活力。在PEA存在的情况下,对酿酒酵母有协同抑制作用,因此水相中PEA的最大浓度仅达到2.1 g/L,而正常补料分批培养时为3.8 g/L。然而,发酵罐中PEA的总浓度增加到了12.6 g/L,因为油相中PEA的浓度达到了24 g/L。因此,在两相补料分批培养中,平均PEA体积生产率为0.26 g L⁻¹ h⁻¹,最大PEA体积生产率为0.47 g L⁻¹ h⁻¹。由于必须避免乙醇抑制,生产率受酿酒酵母固有氧化能力的限制。此外,两相系统的高粘度降低了k(l)a,因此也降低了生产率。因此,如果计划采用特定的ISPR技术,必须记住,在高细胞密度下,该生物转化过程的生产率也会很快受到发酵罐k(l)a的限制。