NEW TECHNIQUE FOR RECOVERY OF PHB FROM RECOMBINANT Escherichia coli BASED ON EXPRESSION OF LYTIC GENES OF PHAGE λ WITH S AMBER MUTATION

YU Huimin; SHI Yue; SUN Xudong; LUO Hui; SHEN Zhongyao

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NEW TECHNIQUE FOR RECOVERY OF PHB FROM RECOMBINANT Escherichia coli BASED ON EXPRESSION OF LYTIC GENES OF PHAGE λ WITH S AMBER MUTATION

更新时间：2025-12-10

- NEW TECHNIQUE FOR RECOVERY OF PHB FROM RECOMBINANT Escherichia coli BASED ON EXPRESSION OF LYTIC GENES OF PHAGE λ WITH S AMBER MUTATION
- CIESC Journal Issue 4, Pages: 623-628(2004)
- 作者机构：
  
  清华大学化学工程系,北京,100084
- 作者简介：
- 基金信息：
- DOI：
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- Online First：25 April 2004，
  
  Published：25 April 2004
- 稿件说明：
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YU Huimin, SHI Yue, SUN Xudong, et al. NEW TECHNIQUE FOR RECOVERY OF PHB FROM RECOMBINANT Escherichia coli BASED ON EXPRESSION OF LYTIC GENES OF PHAGE λ WITH S AMBER MUTATION[J]. CIESC Journal, 2004, (4): 623-628.
DOI：

YU Huimin, SHI Yue, SUN Xudong, et al. NEW TECHNIQUE FOR RECOVERY OF PHB FROM RECOMBINANT Escherichia coli BASED ON EXPRESSION OF LYTIC GENES OF PHAGE λ WITH S AMBER MUTATION[J]. CIESC Journal, 2004, (4): 623-628. DOI：

摘要

将S基因琥珀突变的λ噬菌体裂解基因(S-RRz)引入产聚β-羟基丁酸酯（PHB）的重组大肠杆菌VG1(pTU14)中以实现细胞的可控裂解破壁.采用EDTA/Tris(pH值8.0)缓冲液处理结果表明，S-RRz在VG1(pTU14)中能够成功表达，且EDTA对细胞裂解的决定性作用是由于它模拟了S基因产物的功能.当细胞内PHB含量为85％～90％时，大量积累的PHB颗粒可以改变细胞膜的通透性，实现重组细胞的内控自裂解.对PHB与细胞进行直接分离的后处理工艺研究表明，在S-RRz成功表达的基础上，采用升温处理模拟S基因产物的功能诱导细胞自裂解，PHB产品纯度可以达到95％以上.

Abstract

The lytic genes of phage λ with S amber mutation(S-RRz) were introduced into the recombinant E. coli VG1(pTU14) producing poly-β-hydroxybutyrate(PHB)to attain controllable lysis of cells. The results of EDTA/Tris(pH8.0)buffer treatment showed that S-RRz were successfully expressed in VG1(pTU14)

and cell lysis was realized due to the action of EDTA on cytoplasm membrane.Here the function of EDTA was similar to that of S gene product.When PHB content was 85%—90％，membrane permeability would be increased by the abundantly accumulated in-cell PHB granules

and then the autolysis of recombinant cells occurred. After studies on different projects for direct separation of PHB from fermentation broth

a new technique

in which temperature treatment was introduced to simulate the function of S gene product

was presented

and the autolysis of cells was then easily realized based on the successful expression of S-RRz.By this simple technique

the final purity of PHB product could be up to 95%.

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