Dphydrozyphenylglycine+production+from+DLphydrozyphenylhydantoin+using+the+whole+cells+in+aqueous+twophase+systems研究.pdf

D-p-hydroxyphenylglycine production from DL-p-hydroxyphenylhydantoin using the whole cells in aqueous two-phase systems Bin-Feng Lv Jun-Qing Qian ﹡ Hai-Jun Qian Sheng-Li Yang Xue-Xiao Ying （College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, 310014 China） Abstract: The whole cells bioconversion of DL-p-hydroxyphenylhydantoin (DL-p-HPH) to D-p-hydroxyphenylglycine (D-p-HPG) were carried out in aqueous two-phase system (ATPS) using D-Hydantoinase (D-Hase) and N-carbamoylase (N-Case) from Pseudomonas putida AS 11820. The optimum ATPS consised of PEG 1500 (15%, w/v)/sodium sulfate (15%, w/v). The partition coefficient of DL-p-HPH (KDL-p-HPH) was 28.6, and the partition coefficients of NcHPG (KNcHPG) and D-p-HPG (KD-p-HPG) were 1.45 and 1.25, respectively. When resting -1 cells (0.02g DCW·mL ) was used for bioconversion in ATPS, a highest conversion yield (61%) was achieved at the concentration of 50mM DL-p-HPH as substrate in this ATPS than that of in aqueous system (5mM) at pH9.9 and 28℃. The activity of D-Hase was remained 71% after resting cells was retained in the ATPS for 40 h. Keywords: Pseudomonas putida ; N-carbamoyl-D-p-hydroxyphenylglycine; D-Hydantoinase; Aqueous twp-phase system Introduction Optically active D-amino acids are widely used as intermediates in the pharmaceutic field for the production of semisynthetic antibiotics, peptides, hormones and pesticides [1]. Among them, D-p-hydroxyphenylglycine(D-p-HPG), is used for the synthesis of amoxicillin and cefadroxil. D-specific hydantoinase (dihydroxyprimidinase, EC.3.5.2.2), which is widespread in nature [2], has been used to catalyze the hydro