Vicenzi J T, Hansen G J
Lilly Research Laboratories, Eli Lilly & Co., Lilly Corporate Center, Indianapolis, Indiana 46285.
Enzyme Microb Technol. 1993 Apr;15(4):281-5. doi: 10.1016/0141-0229(93)90150-z.
An economical process for the enzymatic oxidation of cephalosporin C to glutaryl-7-ACA was developed at a pilot plant scale. The process utilized nonviable whole cells of the yeast Triginopsis variabilis containing high levels of D-amino acid oxidase. Prior to use, the whole cells were permeabilized with a 25% acetone/water solution which enhanced their apparent activity by 20- to 50-fold. After permeabilization, the whole cells were incubated at pH 11, which served to selectively deactivate catalase which was present in very large quantities. Deactivation of catalase was critical to achieving high reaction yields. The whole cells were utilized within a "cross-flow filter-reactor" which allowed easy and economical recycle of the cells for repeated use. The overall yield of glutaryl-7-ACA from cephalosporin C was 90-95%. The overall productivity of the yeast was 13 kg cephalosporin C oxidized per kilogram yeast (dry basis). The reaction was run at a concentration of 40 g cephalosporin CL-1 and the overall reactor productivity was 11 g glutaryl-7-ACA l-1 h-1. The process has been thoroughly demonstrated on a 35-l scale, and it should be directly scaleable to 10,000 l or more.
在中试规模下开发了一种将头孢菌素C酶氧化为戊二酰-7-氨基头孢烷酸的经济工艺。该工艺使用了含有高水平D-氨基酸氧化酶的非活性全细胞酵母Triginopsis variabilis。在使用前，用25%的丙酮/水溶液对全细胞进行透化处理，这使其表观活性提高了20至50倍。透化处理后，将全细胞在pH 11下孵育，这有助于选择性地使大量存在的过氧化氢酶失活。过氧化氢酶的失活对于实现高反应产率至关重要。全细胞在“错流过滤反应器”中使用，该反应器便于且经济地回收细胞以供重复使用。从头孢菌素C得到的戊二酰-7-氨基头孢烷酸的总产率为90-95%。酵母的总生产率为每千克酵母（干基）氧化13千克头孢菌素C。反应在40 g头孢菌素C L-1的浓度下进行，反应器的总生产率为11 g戊二酰-7-氨基头孢烷酸l-1 h-1。该工艺已在35升规模上得到充分验证，并且应该可以直接扩大到10000升或更大规模。
