The invention relates to a process for producing three recombinases capable of hydrolyzing organophosphorus,
carbamate and
pyrethroid insecticides, and preparations containing these enzymes. The present invention provides recombinant
plasmid DNA vectors in which the
protein secretion system has been modified to improve the ability of recombinant proteins to secrete into the space around the
plasmid or culture medium. A
nucleotide sequence for coding a PelB (
butyric acid lyase B)
signal sequence is deleted and substituted by a
nucleotide sequence for coding a LamB (
maltodextrin or phage
lambda receptor)
signal sequence, so that the
secretion efficiency of the recombinant
protein is higher. The invention discloses three recombinant
plasmid DNA (deoxyribonucleic acid) vectors which are constructed by utilizing the improved
plasmid DNA vectors and are used for expressing recombinant proteins. The recombinant
protein comprises (a) a recombinant
organophosphate hydrolase having at least 80%
sequence identity with the
amino acid sequence of
methyl parathion hydrolase (MPH) (
Protein Database (PDB) ID: 1P9E) as shown in SEQ ID NO: 1, wherein the recombinant
organophosphate hydrolase has hydrolytic activity on
organophosphate insecticides; (b) a recombinant
carbamate hydrolase having at least 80%
sequence identity with the
amino acid sequence of L-
aminopeptidase-D-Ala-
esterase /
amidase (
aryl amidase) (MAH) (PDB ID: 1B65), as shown in SEQ ID NO: 2, wherein the recombinant
carbamate hydrolase has hydrolytic activity on carbamate insecticides; and (c) a recombinant
pyrethroid hydrolase having at least 80%
sequence identity with the
amino acid sequence of the
pyrethroid hydrolytic
carboxylesterase (EstA) (PDB ID: 4CCY) as shown in SEQ ID NO: 3, wherein the recombinant pyrethroid hydrolase has hydrolytic activity on a
pyrethroid insecticide. The embodiment of the invention further provides three recombinant bacterial strains, an
escherichia coli strain BL21 (DE3) is used as a host
cell, and each recombinant
plasmid DNA vector is used for transformation. The invention also discloses culture conditions of the recombinant strain, wherein the culture conditions comprise culture medium composition, induction conditions and
enzyme harvesting and
enzyme activity detection methods. In addition, the embodiment of the invention also provides a preparation which contains the three recombinases as functional components.