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.
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:
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%.