Conlon H D, Baqai J, Baker K, Shen Y Q, Wong B L, Noiles R, Rausch C W
Biopure Corporation, 11 Hurley Street, Cambridge, MA 02141, USA.
Biotechnol Bioeng. 1995 Jun 20;46(6):510-3. doi: 10.1002/bit.260460603.
The first large-scale production of 7-aminocephalosporanic acid (7ACA) from cephalosporin C (CPC) using a wholly enzymatic synthesis method is reported here. We produced 7ACA from CPC in as high a molar yield as 85% using the immobilized enzymes D-amino acid oxidase (D-AOD) and glutaryl-7-ACA acylase (GL-acylase). In the first reactor, CPC is converted to keto-adipyl-7-aminocephalosporanic acid (keto-7ACA) using an immobilized D-AOD isolated from a yeast, Trigonopsis variabilis. The keto-7ACA is then spontaneously converted to glutaryl-7-aminocephalosporanic acid (GL-7ACA) via a chemical reaction with hydrogen peroxide. The hydrogen peroxide is also a product of the D-AOD reaction. Near quantitative conversion of the keto-7ACA to GL-7ACA was observed. The second reactor converts GL-7ACA to 7ACA using an immobilized GL-acylase, which was isolated from a recombinant Escherichia coli. The final 7ACA crystalline product is a high quality product. The reactions are conducted under very mild aqueous conditions: pH 8.0 and 20 degrees to 25 degrees C. The production of desacetyl side products is minimal. This process is currently being implemented on an industrial scale to produce 7ACA.
本文报道了首次采用全酶法从头孢菌素C（CPC）大规模生产7-氨基头孢烷酸（7ACA）的情况。我们使用固定化酶D-氨基酸氧化酶（D-AOD）和戊二酰-7-ACA酰基转移酶（GL-酰基转移酶），以高达85%的摩尔产率从CPC生产出了7ACA。在第一个反应器中，使用从可变三角酵母中分离得到的固定化D-AOD将CPC转化为酮己二酰-7-氨基头孢烷酸（酮-7ACA）。然后，酮-7ACA通过与过氧化氢的化学反应自发转化为戊二酰-7-氨基头孢烷酸（GL-7ACA）。过氧化氢也是D-AOD反应的产物。观察到酮-7ACA几乎定量转化为GL-7ACA。第二个反应器使用从重组大肠杆菌中分离得到的固定化GL-酰基转移酶将GL-7ACA转化为7ACA。最终的7ACA结晶产物质量很高。反应在非常温和的水性条件下进行：pH 8.0，温度20摄氏度至25摄氏度。脱乙酰副产物的产生极少。该工艺目前正在工业规模上实施以生产7ACA。
