| 青霉素G酰化酶通用表达载体的构建方法初探 |
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| 引用本文： | 陈美娟,周政,袁中一.青霉素G酰化酶通用表达载体的构建方法初探[J].药物生物技术,2003,10(4):215-217. |
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| 作者姓名： | 陈美娟 周政 袁中一 |
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| 作者单位： | 1. 南京中医药大学,江苏,南京,210029
2. 中科院上海生物化学研究所,上海,200031 |
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| 摘 要： | 结合信号肽预测软件(Signal P)对现有的青霉素G酰化酶(PGA)的大肠杆菌-枯草杆菌穿梭表达质粒pZOl进行信号肽的合理设计,在其C-端置换2个氨基酸,即引入NotI酶切位点序列,从而构建成PGA的通用表达载体。将粪产碱杆菌(Alcaligenes faecalis)的PGA基因导入其中,构建成表达质粒pefpganoti．,转入枯草杆菌进行表达,测活结果显示,表达产物具有生物活性(酶的平均活力为0．18U／ml发酵液)。为进一步应用信号肽预测软件进行表达系统的研究、设计打下了基础。
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| 关 键 词： | 信号肽 青霉素G酰化酶 粪产碱杆菌 PGA基因 基因表达 半合成β一内酰胺类抗生素 |
| 文章编号： | 1005-8915(2003)04-0215-03 |
Expression System Construction of the Gene Encoding Penicillin G Acylase |
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| CHEN Mei-juan,ZHOU Zheng,YUAN Zhong-yi.Expression System Construction of the Gene Encoding Penicillin G Acylase[J].Pharmaceutical Biotechnology,2003,10(4):215-217. |
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| Authors: | CHEN Mei-juan ZHOU Zheng YUAN Zhong-yi |
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| Institution: | CHEN Mei-juan1,ZHOU Zheng2,YUAN Zhong-yi2 |
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| Abstract: | In order to construct a new expression vector which can excrete penicillin G acylase, we redesigned the signal peptide sequence of the penicillin G acylase from Bacillus megterium with the "Signal P" software .At the c-terminal of the signal peptide two amino acids were changed and a new enzyme cut site "NotI" was created.Then the gene encoding the penicillin G acylase from Alcaligenes faecalis was cloned into the new vector.The average value of the excreted enzyme was 0.18U/ml.This means the way of "Signal P" deduction could be used to help designing the expression vector. |
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| Keywords: | Signal Peptide Penicillin G acylase Alcaligenes faecalis |
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