D-对羟基苯海因细菌的发酵工艺优化研究

The Optimization of Fermentation Process of DL-Hydroxyphenyl Hydantoin Bacterial

通过筛菌得到一株野生菌LLV-6,在诱导剂诱导后利用DL-对羟基苯海因(DL-HPH)为底物经D-海因酶(HYD)和N-氨甲酰-D-氨基酸酰胺水解酶(CAB)两酶水解而获得D-对羟基苯甘氨酸。用LB培养基(含磷酸盐缓冲溶液,pH=7.2)接菌量为0.5%(对数期的菌种),在37℃、200r、pH7.2条件下培养菌种。在培养12h后加入2%的诱导剂并以500μLDMSO作为促溶剂,33h后收集细胞。通过对细胞酶活力的测定,此时酶活力达到最大值,酶活力为0.57U/L,比优化前提高了2.6倍。

Through the sieve bacteria get one natural bacteria LLV-6,In inducers induced effect after using the DL-Hydroxyphenyl Hydantoin（DL-HPH） as the substrate by D-hydantoinase（HYD）and N-carbamoylase （CAB）two enzyme hydrolysis of D-p-Hydroxyphenylglycine（D-HPG）. Using LB culture medium（containing phosphate buffer solution,pH=7.2）the inoculation quantity was 0.5%（Logarithmic phase）,the bacteria cul-tured at 37℃,200r,pH7.2 conditions. In 12 hours after adding 2% inducers and with DMSO as solvent,33 hours after collection of the cell. By measuring the enzyme activity of the cells,the enzyme activity reached a maximum value at this time,Enzyme activity was 0.57U/L,more than 2.6 times before optimiza-tion.