Activation of a Bacterial Lipase by its Chaperone

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 12; pp. 5682 - 5686
• Main Authors: Hobson, Audrey H., Buckley, Catherine M., Aamand, Jesper L., Jorgensen, Steen T., Diderichsen, Borge, McConnell, David J.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.06.1993; National Acad Sciences; National Academy of Sciences
• Subjects: Amino acids; Antibodies; Bacteria; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biological and medical sciences; Biotechnology; Burkholderia cepacia - enzymology; Burkholderia cepacia - genetics; Cell membranes; Cloning, Molecular; Enzyme Activation; Enzyme Precursors - metabolism; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes; Genes, Bacterial; Genes, Regulator; Genetics; Immunoprecipitation; Kinetics; Lipase - biosynthesis; Lipase - genetics; Lipase - metabolism; Molecular and cellular biology; Molecular chaperones; Molecular genetics; Plasmids; Protein Biosynthesis; Protein Sorting Signals - metabolism; Proteins; Pseudomonas cepacia; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; RNA, Messenger - metabolism; Secretion; Transcription, Genetic; Translation. Translation factors. Protein processing; Pseudomonadales; Heterologous system; Molecular processing; Enzyme; Gene product; Chaperone; Triacylglycerol lipase; Activation; Gene expression; Precursor; Pseudomonas cepacia; Bacteria; Pseudomonadaceae; Molecular complex
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.12.5682

The gene lipA of Pseudomonas cepacia DSM 3959 encodes a prelipase from which a signal peptide is cleaved during secretion, producing a mature extracellular lipase. Expression of lipase in several heterologous hosts depends on the presence of another gene, limA, in cis or in trans. Lipase protein has been overproduced in Escherichia coli in the presence and absence of the lipase modulator gene limA. Therefore, limA is not required for the transcription of lipA or for the translation of the lipA mRNA. However, no lipase activity is observed in the absence of limA. limA has been overexpressed and encodes a 33-kDa protein, Lim. If lipase protein is denatured in 8 M urea and the urea is removed by dialysis, lipase activity is quantitatively recovered provided Lim protein is present during renaturation. Lip and Lim proteins form a complex precipitable either by an anti-lipase or anti-Lim antibody. The Lim protein has therefore the properties of a chaperone.