摘要
以豇豆褪绿斑驳病毒(CCMV)衣壳蛋白表达基因为模板,构建基因工程菌CCMV-BL21,并通过正交实验优化可溶蛋白表达工艺;采用原位还原法制备病毒样颗粒限域Ru纳米催化剂(Ru@CCMV),以肉桂醛和4-硝基苯酚加氢为模型反应进行评价。目标蛋白的最佳表达条件为:诱导温度25℃,异丙基硫代半乳糖苷(IPTG)终浓度1.0 mmol/L,诱导时间15 h。在该条件下,可溶性蛋白表达量达181 mg/L发酵液,约占表达蛋白总质量的93%。Ru@CCMV在两个反应中均显示较高的催化活性,反应速率常数经计算分别达到0.17 min^-1和0.50 h^-1,高于柠檬酸修饰Ru纳米颗粒(Ru-CA)催化剂(0.11 min^-1和0.36 h^-1)。
A genetically engineered bacteria CCMV-BL21 was firstly constructed using the gene expressing cowpea chlorotic mottle virus(CCMV) capsid proteins(CPs) as template. The expression process of soluble CCMV CPs was then optimized by orthogonal experiment. Subsequently, Ru nanoparticles encapsulated in CCMV CPs(Ru@CCMV) self-assembled virus-like particles(VLPs) was prepared by in-situ reduction. The catalytic activity of the prepared catalyst Ru@CCMV was investigated by catalytic hydrogenation of cinnamaldehyde(CAL) and 4-nitrophenol(4-NP), respectively. The results showed that the optimal expression conditions of CCMV CPs were as follows: induction temperature 25 ℃, final concentration of IPTG 1.0 mmol/L and induction time 15 h. Under the optimal conditions, the content of expressed soluble CCMV CPs reached 181 mg/L fermentation broth, accounting for 93% of the total protein expression. Ru@CCMV exhibited good catalytic activity in both reactions. The corresponding reaction rate constants over Ru@CCMV were calculated to be 0.17 min^-1 and 0.50 h^-1, respectively, higher than those over Ru nanoparticles modified by citric acid(Ru-CA)(0.11 min^-1 and 0.36 h^-1).
作者
朱劼
胡加慧
杨坤
卢晓雪
ZHU Jie;HU Jia-Hui;YANG Kun;LU Xiao-Xue(School of Pharmaceutical Engineering&Life Science,Changzhou University,Changzhou 213164,Jiangsu,China)
出处
《精细化工》
EI
CAS
CSCD
北大核心
2019年第11期2227-2233,共7页
Fine Chemicals
基金
国家自然科学基金（21676029)
关键词
病毒样颗粒
豇豆褪绿斑驳病毒
原核表达
纳米催化剂
催化技术
virus-like particle
cowpea chlorotic mottle virus
prokaryotic expression
nanocatalyst
catalysis technology
作者简介
朱劼(1977-),男,副教授,E-mail:zhujie@cczu.edu.cn。
