科研之友
微信
新浪微博
Facebook
分享链接
摘要
Desaturases, key enzymes in the metabolism of fatty acids, regulate the physical and biochemical properties of membranes. They adjust the composition of saturated and unsaturated fatty acids in response to changes in the environmental. We demonstrated the existence of Delta 9 desaturase activity in epimastigotes of the Trypanosoma cruzi Tulahuen strain. In the present study, showed that this enzyme has an approximate molecular mass of 50 kDa and a pl value of approximately 9. In order to characterize the Delta 9 desaturase of Trypanosoma cruzi, (Tc Delta 9DES) we have cloned, sequenced and expressed in Escherichia coli. The gene consists of 1300 by and encodes a peptide of 433 amino acids with a molecular weight of 50 kDa. Analysis of the amino acid sequence revealed three clusters of histidine and two hydrophobic regions, characteristic of membrane bound desaturases. Gene expression studies showed that Tc Delta 9DES was overexpressed as an active protein. Fatty acid analysis showed that the expressed protein was confirmed to be functional with Delta 9 desaturase activity. This enzyme changed the fatty acid profile of Tc Delta 9DES-expressing E. coli, decreasing the levels of palmitic (16:0) and stearic (18:0) acids and enhancing palmitoleic (16:1 Delta 9) and monounsaturated 18 carbons fatty acids. When [1-14C]palmitic or [1-14C]stearic acid was used as substrate, TcA9DES-expressing E. coli exhibited high desaturase activity associated with increased levels of monounsaturated fatty acids, suggesting that the Tc Delta 9DES enzyme was actively expressed in E. coli. To check the commitment of Tc Delta 9DES against sterol biosynthesis inhibitors we tested the activity under ketoconazole effect. Native Tc Delta 9DES, showed a significant activity inhibition. Since Tc Delta 9DES has shown active
- 出版日期2017
