Full Length Research Paper

Study on optimal production of 3-ketovalidoxylamine A C-N lyase and glucoside 3-dehydrogenase by a newly isolated Stenotrophomonas maltrophilia

Jian-Fen Zhang¹, Yu-Guo Zheng²* and Yin-Chu Shen²

¹College of Biology and Environmental Engineering, Zhejiang Shuren University, Hangzhou 310015, People’s Republic of China.

²Institute of Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People’s Republic of China.

*Corresponding author. E -mail: zhengyg@zjut.edu.cn. Tel: +86-571-88320630. Fax: +86-571-88320630.

Accepted 10 September, 2009

   Abstract

3-Ketovalidoxylamine A C-N lyase and glucoside 3-dehydrogenase (G3DH), two key enzymes for valienamine synthesis, are produced by Stenotrophomonas maltrophilia. The condition of producing 3-ketovalidoxylamine A C-N lyase and G3DH was optimized. Validamycin A was showed to be suitable carbon source for C-N lyase and G3DH synthesis. Maximum C-N lyase production was achieved by adding 100 ml culture medium into a 500 ml conical flask containing 5 mg/ml validamycin A, with 12% inoculation seed volume for 22 h with a productivity of 6.26 × 10^-3 U/mg protein. The specific G3DH activity was 0.244 U/mg protein in the same culture condition at 27 h. A 15 L fermenter containing 8 liter of medium was also used in this study. Crude enzyme reaction showed the conversion of N-p-nitrophenylvalidamine was achieved to 92.8% at 3 h and the yield of p-nitroaniline was 81.3% at 3 h.

Key words: 3-Ketovalidoxylamine A C-N lyase, glucoside 3-dehydrogenase, validamycin A, valienamine.