阿莫西林酰化酶发酵工艺的研究  

Optimization on the Fermentation Process of Amoxicillin Acylase

作　　者：李希平 王娟 LI Xiping;WANG Juan(Hangzhou Xianghu High-tech Application Research Institute,Hangzhou 311200,China;Standard Sci-Tech Innovation(Qingdao)Pharmaceutical Technology Co.,Ltd.,Qingdao 26614,China)

机构地区：[1]杭州市萧山区湘湖高新技术应用研究院,浙江杭州311200 [2]斯坦德科创医药科技(青岛)有限公司,山东青岛266114

出　　处：《药物生物技术》2025年第1期39-43,共5页Pharmaceutical Biotechnology

摘　　要：阿莫西林作为一种较为常见的抗菌类药物,是世界卫生组织(WHO)推荐的首选口服类β-内酰胺类抗生素。自1972年上市以来,阿莫西林因其抗菌高效广谱且毒副作用小,在临床医学被广泛应用,2023年中国医药工业信息中心数据显示全球阿莫西林原料药年需求约30000吨,市场巨大。在其应用长达半个世纪后仍保持着强劲的生命力,因此优化阿莫西林的制备技术有重要意义。目前阿莫西林已经形成了相对较为成熟的工业化生产工艺,主要采用的工艺技术方法有两种,分别是化学法和酶法。其中酶法的工艺优势要高于化学法,是目前制备阿莫西林的主流制备工艺。合成阿莫西林的酰化酶因其为阿莫西林制备工艺的关键酶,其质量直接决定着成品的质量与成本。本研究根据仿制药研发注册要求,分别对合成阿莫西林的酰化酶发酵使用的培养基,发酵过程的关键参数pH、温度、IPTG添加时机、摇床转速、抗生素添加等方面进行了实验设计优化。结果显示在种子培养阶段添加抗生素,将培养基中Yeast extract、Tryptone替换成酵母粉FM902和酵母蛋白胨FP101并调节培养基初始pH值至6.60,接种培养2 h加入IPTG诱导,诱导后28℃、130 r/min继续培养14 h获得的酶活最高。优化后酶活较先前的报道[1]提高约64%,较实验室的初始发酵方法提高约95%。本研究为优化阿莫西林的制备技术提供了重要的借鉴。Amoxicillin,as a common antibacterial drug,is the preferred oralβ-lactamide antibiotics recommended by the World Health Organization(WHO).Since its launch in 1972,amoxicillin has been widely used in clinical medicine due to its high antibacterial efficiency,broad-spectrum efficacy,and low toxic side effects.According to data from the China Medical Industry Information Center in 2023,the global demand for amoxicillin is about 30000 tons per year,with a huge market.After being applied for half a century,it still maintains strong vitality,so optimizing the preparation technology of amoxicillin is still of great significance.At present,it has formed a relatively mature industrial production process for amoxicillin,mainly using two technological methods:Che-mical method and enzymatic method.The advantage of enzymatic method is higher than chemical method,which is currently the mainstream preparation process for amoxicillin.The acylase for synthesizing amoxicillin is a key enzyme in the amoxicillin preparation process,and its quality directly determines the quality and cost of the finished product.This study optimized the experimental design of the fermentation medium used for acylase synthesis of amoxicillin,as well as the key parameters of the fermentation process,such as pH,temperature,timing of IPTG addition,shaking speed,and antibiotic addition,in accordance with the requirements for generic drug development registration.The results showed that the highest enzyme activity was obtained by adding antibiotics during the seed cultivation stage,replacing yeast extract and tryptone with yeast powder FM902 and yeast peptone FP101 in the medium,adjusting the initial pH of the medium to 6.60,adding IPTG after 2 hours of inoculation and cultivation,continuing cultivation at 28℃and 130 r/min for 14 hours.After optimization,the enzyme activity increased by about 64%compared to previous report,and by about 95%compared to the initial fermentation method in the laboratory.This study provides important reference for optimizing

关 键 词：阿莫西林 酰化酶 培养基 温度 PH 异丙基-β-D-硫代半乳糖苷 转速 酶活 

分 类 号：Q814[生物学—生物工程] TQ460.1[化学工程—制药化工]