Production of active Pseudomonas glumae lipase in homologous or

Details Production of active Pseudomonas glumae lipase in homologous or Production of active Pseudomonas glumae lipase in homologous or

1995-05-24

1998-09-08

Railey, II, Johnny F.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435198, 4352523, 43525234, 436 232, C12N 121, C12N 1531, C12P 2102, C07K 14195

Patent

active

058044094

ABSTRACT:
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 optimising 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.

REFERENCES:
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patent: 4551433 (1985-11-01), DeBoer
patent: 4897471 (1990-01-01), Stabinsky
Jorgensen et al., "Cloning, Sequence & Expression of a Lipase Gene from Pseudomonas cepacia: Lipase Production in Heterologous Hosts Requires Two Pseudomonas Genes", Journal of Bacteriology vol. 173 No. 2, Jan. 1991, pp. 559-567.
Biotechnology Abstracts Database abstract No. 89-02896, 1989, Derwent Pub. Ltd., London, GB; Aoyama et al.: "Cloning, sequencing and expression of the lipase gene from pseudomonas fragi IFO-12049 in E. coli",vol. 242, pp. 36-40.
Chihara-Siomi et al., Arch. of Biochem. & Biophys., vol. 296, No. 2, pp. 505-513 (1992).
Reznikoff et al., "E. coli Promoters", pp. 1-4 (1986); in Maximizing Gene Expression, Butterworth Publ., Stoneham, MA.

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