Bushueva A M, Shevelev A B, Gumpert J, Chestukhina G G, Serkina A V, Hoischen C, Matz M V, Kuryatova M V, Stepanov V M
Laboratory of Protein Chemistry, Institute of Microbial Genetics (GNII Genetika), Moscow, Russia.
FEMS Microbiol Lett. 1998 Feb 15;159(2):145-50. doi: 10.1111/j.1574-6968.1998.tb12853.x.
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.
