PURPOSE: To produce lipase in high efficiency, by using a recombinant DNA replicable in the cell of Escherichia coli and obtained by integrating a lipase structure gene originated from Gram-negative bacteria to a DNA-introducing vector for Escherichia coli.
CONSTITUTION: A phage-vector is integrated into a Gram-negative bacterium such as lipase-producing Alcaligenes genus, etc., and the chromosome DNA of the Gram-negative strain is inserted in the phage vector. The phage vector containing the lipase structure gene is introduced to the cell of second bacterial strain free from the lipase-producing capability. The second strain is selected according to the marker and the lipase-productivity of the above phage vector, and the whole DNA is extracted from the strain and fragmentized. The fragment is integrated to a multicopy-type plasmid vector replicable in Escherichia coli, and the recombinant DNA containing the lipase structure gene having lipase- producing capability is selected from the obtained recombinant DNA. Escherichia coli is transformed with the resultant DNA, and is cultured to produce lipase.
NEGI TAHEE
HIGASHIE AKIO
