|
|||||
|
|
Expression of the carboxypeptidase T gene from Thermoactinomyces vulgaris in stable protoplast type L-forms of Proteus mirabilis |
|
| Authors: | A.M Bushueva A.B Shevelev J Gumpert G.G Chestukhina A.V Serkina C Hoischen M.V Matz M.V Kuryatova V.M Stepanov |
| Affiliation: | Laboratory of Protein Chemistry, Institute of Microbial Genetics (GNII Genetika), 1st Dorozhny, 1 Moscow 113545, Russia;Molecular Biology Department, Biological Faculty, Moscow State University, Moscow, Russia;Laboratory of Prokaryotic Cell Biology, Institute of Molecular Biotechnology, Jena, Germany |
| Abstract: | The structural gene of the carboxypeptidase T (cpt) was successfully expressed in cell wall-less L-form cells of Proteus mirabilis. The DNA sequence encoding the PhoA leader peptide was fused with a truncated cpt gene encoding the mature enzyme. The modified gene in a pUC-based kanamycin resistance vector under the control of the lac promoter was transformed into L-form cells of P. mirabilis. They were able to produce the recombinant CpT both as a secretory and as a cell-bound insoluble form. The co-secretory processing of the PhoA leader peptide was quite efficient. The yield of the secreted CpT was not less than 20 mg l−1 and should be improvable. |
| Keywords: | Secretion Propeptide Carboxypeptidase L-form Proteus mirabilis |
| 本文献已被 ScienceDirect 等数据库收录！ | |