The Application of a Novel Lytic System to the Recovery of Recombinant Proteins in E.coli.
In order to efficiently recover recombinant proteins, a temperature-sensitive lytic system was constructed on the basis of the feature that T4 lysozyme disrupts the bacteria through cutting specific bond in the peptidoglycan layer of cell wall. This system was evaluated by constructing and introducing a low copy plasmid pSC-lys (pSC101 replication origin) into E.coli. The plasmid contained a temperature sensitive T4 lysozyme (LYS(ts)) gene under the control of three tandem tac promoters and the LacI repressor, which is compatible with other plasmids carrying pMB1, ColE1 replication origins, etc. Under the optimum lysis conditions, 2--5 fold condensed cultures resuspended in buffer A, beta-galactosidase, recombinant chaperone GroEL and ZZ-fusion salmon hexamic calcitonin (Cal6) in E.coli were released simply, rapidly, and quantitatively, as co-expressed with LYS(ts). The two tested recombinant proteins maintained their significant productions. Instead of other cumbersome lysising methods, this novel lytic system will be useful in recovery of recombinant proteins for further purification in the field of biotechnology.
Yang
Yun-Gui
YG
Shanghai Research Center of Biotechnology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200233, China. ygong@srcb.ac.cn
Tong
Qin
Q
Hu
Tai-Shan
TS
Qian
You-Cun
YC
Yang
Sheng-Li
SL
Gong
Yi
Y