生物聚合物聚-3-羟基丁酸酯的酶法提取纯化研究  

Study on enzymatic extraction and purification of bio-based poly-3-hydroxybutyrate

作　　者：薛英茹 周浩 周卫强 刘海军 李义 李非[1] XUE Ying-ru;ZHOU Hao;ZHOU Wei-qiang;LIU Hai-jun;LI Yi;LI Fei(School of Chemical Engineering and Technology,Hebei University of Technology,Tianjin 300401,China;Nutrition and Health Research Institute,COFCO,Beijing 102209,China;National Engineering Research Center of Corn Deep Processing,Jilin COFCO Biochemistry Co.,Ltd.,Changchun 130033,China;COFCO Biomaterial(Yushu)Co.,Ltd.,Changchun 130401,China)

机构地区：[1]河北工业大学化工学院,天津300401 [2]中粮营养健康研究院有限公司,北京102209 [3]吉林中粮生化有限公司,玉米深加工国家工程研究中心,吉林长春130033 [4]中粮生物材料(榆树)有限公司,吉林长春130401

出　　处：《现代化工》2025年第1期137-142,共6页Modern Chemical Industry

基　　金：科技部重点研发计划项目(2021YFC2101702);吉林省科技发展计划项目(212551GX010489094)。

摘　　要：通过酶解法从嗜盐菌发酵液中获得高纯度、高分子质量的聚-3-羟基丁酸酯(PHB),分别采用高温处理与胞壁酸酶酶解2种方法对嗜盐菌细胞进行破壁,再通过中碱性蛋白酶进一步酶解纯化。结果表明,121℃下高温处理可有效实现嗜盐菌细胞破壁,但高温环境使PHB分子质量降低了30%~40%;在50℃、pH 6条件下添加0.3%胞壁酸酶酶解60 min破壁效果最好,且分子质量可维持在10%以内;在70℃、pH 10条件下添加0.045%中碱性蛋白酶酶解60 min纯化效果最好,所得PHB纯度可达97%以上。最后与市售PHB进行对比,确认了所得PHB的化学结构及热性能。酶的特异性水解保证了PHB结构不受破坏,胞壁酸酶-中碱性蛋白酶结合酶解法可以得到高纯度、高分子质量的PHB,无需其他化学试剂辅助。Poly-3-hydroxybutyrate(PHB)with high purity and high molecular weight is obtained from halophiles fermentation broth by enzymatic digestion method.Cell walls of halophiles are separately broken by heat treatment and muramidase hydrolysis,and then further purified by neutral alkaline protease.Results show that high temperature treatment at 121℃ can effectively break the walls of halophile cells,but it also makes the molecular weight of PHB decline by 30%-40%.Under the conditions of 50℃ and a pH of 6,the addition of 0.3 wt% muramidase for 60 min of enzymatic hydrolysis can receive the best wall-breaking effect,and the decline of PHB molecular weight can be maintained within 10%.Under the conditions of 70℃ and a pH of 10,adding 0.045 wt% neutral alkaline protease for 60 min of enzymatic hydrolysis can achieve the best purification effect,the resulted PHB has a purity of higher than 97%.Through comparing with commercially available PHBs,the chemical structure and thermal properties of the resulted PHB are confirmed.Specific enzymatic hydrolysis ensures that PHB structure is not destroyed.Therefore,PHB with high purity and high molecular weight can be obtained by muramidase-neutral alkaline protease enzymatic digestion,which is an efficient and environmentally friendly method without the aid of other chemical reagents.

关 键 词：聚-3-羟基丁酸酯 高温处理 酶解 提取 纯化 

分 类 号：TH3[机械工程—机械制造及自动化]