Original Paper

World Journal of Microbiology and Biotechnology

, 26:145

First online:

Single-pot conversion of cephalosporin C to 7-aminocephalosporanic acid in the absence of hydrogen peroxide

  • Qiang TanAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and TechnologyFaculty of Pharmacy, Guangxi Traditional Chinese Medical University
  • , Yewang ZhangAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and TechnologyFaculty of Pharmaceutical Engineering Department, Jiangsu University
  • , Qingxun SongAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology
  • , Dongzhi WeiAffiliated withState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and TechnologyInstitute of Biochemistry, East China University of Science and Technology Email author 

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Abstract

In this study, d-amino acid oxidase (DAAO) and catalase (CAT) in the permeabilized recombinant Pichia pastori cells were well investigated. It appeared that their thermal stability was negatively correlated with the apparent enzymatic activities. The frozen-melted cells presented the best stability and the lowest apparent activities of DAAO and CAT, whereas the cetyltrimethylammonium bromide (CTAB) permeabilized cells displayed the weakest stability and the highest apparent activities of the two enzymes. Simultaneous action of DAAO and CAT in the CTAB-permeabilized cells and glutaryl-7-aminocephalosporanic acid acylase (GA) immobilized on carrier contributed to the conversion of cephalosporin C (CPC) to 7-aminocephalosporanic acid (7-ACA) with a yield of 76.2%. During such a reaction cycle, no visible activity loss occurred at the immobilized GA, whereas the loss rates of DAAO and CAT activities were about 0.029 and 1.13 U min−1, respectively. Nevertheless, this problem could be easily solved by continuous feeding of the new permeabilized cell suspension at the rate of 6 ml h−1 to the reactor. Following such a fed-batch strategy, these permeabilized cells and the immobilized GA could be efficiently reused for 6 and 15 reaction cycles, respectively, yielding around 76% 7-ACA at each reaction cycle.

Keywords

Cephalosporin C d-Amino acid oxidase Glutaryl-7-aminocephalosporanic acid acylase Catalase 7-Aminocephalosporanic acid