Production of active Pseudomonas glumae lipase in homologous or heterologous hosts

摘要

The production of an active Pseudomonas glumae lipase isolated from P. glumae PG1 (CBS 322.89) and other lipases suitable for application in detergent systems is described in homologous, but particularly in heterologous hosts, e.g. Bacillus subtilis. For the latter a "helper function" or "lipase-specific stabilization/translocation protein" is needed, for which a gene is provided which, when expressed in concert with the lipase gene, can improve the stabilization of the intermediates involved in the production and translocation/secretion of the lipase. The hosts are transformed by recombinant DNA methods and modified lipases can be made by site-directed mutation or classical mutation techniques. The lipase gene and the gene encoding the helper function can be part of one operon that can be transcribed to form a polycistronic messenger or be present as separate genes yielding two mRNA's on transcription. The production level can be further improved by optimizing the regulation sequences. It can be advantageous to use at least part of the signal sequence of the lipase gene in addition to a signal sequence homologous to the host in which the lipase is produced.