基金项目:(B2014208121),河北科技大学大学生创新创业训练计划项目(201510082017),河北科技大学五大平台开放基金(2015PT37)
通过对青霉素G酰化酶(PGA)催化合成头孢丙烯的工艺条件进行优化,得到相关实验数据,为头孢丙烯绿色生产技术的推行打下基础。该优化工艺是以7-氨基-3-[(Z)-烯丙基-1-基]-3-头孢环-4-羧酸(7-APCA)为底物,D-对羟苯甘氨酸甲酯(D-HPGME)为酰基供体,水作溶剂,固定化青霉素G酰化酶为催化剂合成头孢丙烯。筛选合适的催化剂酶活并分别采用氨基和环氧载体固载,考察反应pH值、温度、投酶量、投料比、底物浓度等多个因素对酶法合成头孢丙烯的影响。结果显示:以氨基载体合成头孢丙烯的较适宜条件为pH=6.0~6.5;温度25℃;投酶量/7-APCA(质量比)=1.2:1,D-HPGME:7-APCA=1.2:1;底物浓度500mmol/L。此工艺条件下合成水解比S/H可达5以上,收率为63-72%。以环氧载体合成头孢丙烯的较适宜合成条件为温度pH=6.0~6.5;25℃;投酶量/7-APCA(质量比)=1.3:1,投料比HPGM:7-APCA=1.1:1;底物浓度500mmol/L。此工艺条件下合成水解比S/H可达8以上,收率为76-80%。
TheoptimizationexperimentsofsynthesizingcefprozilwithpenicillinGacylaseascatalystweresuccessfullyachieved,whichcanlaythefoundationofthegreenproductiontechnologyofcefprozil.Theenzymaticsynthesisofcefprozilwasoperatedwith7-amino-3-cephem-4-carboxylicacid(7-APCA)asthemothernucleus,D-hydroxylmethylphenylglycine(D-HPGME)asacyldonor,waterassolventandimmobilizedpenicillinGacylaseasenzymaticcatalyst.TheinfluencefactorsofenzymaticSsynthesisofcefprozilsuchaspH,temperature,enzymedosage,andfeedratiosubstrateconcentrationwereresearchedundertheconditionofappropriateenzymeactivitywithaminocarrierandepoxycarrier.Theresultrevealedthat,morethan5ofhydrolysisratioandyieldof63-75%wereachievedusingaminocarrierwithpHvalueof6.0~6.5,thetemperatureof25℃,themassratioofenzyme:7-APCA(1.2:1)andHPGM:7-APCA(1.2:1),andsubstrateconcentrationof500mmol/L.Undersimilarconditions,whenpHvalueis6.0~6.5,thetemperatureis25℃,themassratioenzymeand7-APCAis1.3:1andHPGM:7-APCA=1.1:1,thesubstrateconcentrationis500mmol/L,morethan8ofsynthetichydrolysisratioandtheyieldof76-80%wereobtainedusingepoxycarrier.