CLONING AND CHARACTERIZATION OF THE LIPASE OPERON FROM MYCOPLASMA-MYCOIDES SUBSPECIES MYCOIDES LC

Lipases, serine esterase enzymes, play an essential role in the mycopl asmal nutritional requirement for long-chain fatty acids. Although the lipase(s) activity in different mycoplasma species has been investiga ted, the molecular biology of the corresponding genes has not been stu died. Using a single-primer PCR technique combined to more classical c loning systems, an operon containing three open reading frames (ORF), each of which could encode a lipase protein of 264, 264 or 269 amino a cids (aa), was identified from Mycoplasma mycoides subsp. mycoides LC. Analysis of aa sequences of the encoded polypeptides showed that they display high aa similarity between each other (65-79%) and 28-31% ide ntity to other prokaryotic lipases. Moreover, a lipase-esterase activi ty could be detected when the mycoplasmal lipase-encoding genes were e xpressed in a strong opal-suppressor-bearing Escherichia coli strain.