| Title : The Role of Residues 103, 104, and 278 in the Activity of SMG1 Lipase from Malassezia globosa: A Site-Directed Mutagenesis Study - Lan_2015_J.Microbiol.Biotechnol_25_1827 |
| Author(s) : Lan D , Wang Q , Popowicz GM , Yang B , Tang Q , Wang Y |
| Ref : J Microbiol Biotechnol , 25 :1827 , 2015 |
| Abstract : The SMG1 lipase from Malassezia globosa is a newly found mono- and diacylglycerol (DAG) lipase that has a unique lid in the loop conformation that differs from the common alpha-helix lid. In the present study, we characterized the contribution of three residues, L103 and F104 in the lid and F278 in the rim of the binding site groove, on the function of SMG1 lipase. Sitedirected mutagenesis was conducted at these sites, and each of the mutants was expressed in the yeast Pichia pastoris, purified, and characterized for their activity toward DAG and pnitrophenol (pNP) ester. Compared with wild-type SMG1, F278A retained approximately 78% of its activity toward DAG, but only 11% activity toward pNP octanoate (pNP-C8). L103G increased its activity on pNP-C8 by approximately 2-fold, whereas F104G showed an approximate 40% decrease in pNP-C8 activity, and they both showed decreased activity on the DAG emulsion. The deletion of 103-104 retained approximately 30% of its activity toward the DAG emulsion, with an almost complete loss of pNP-C8 activity. The deletion of 103-104 showed a weaker penetration ability to a soybean phosphocholine monolayer than wild-type SMG1. Based on the modulation of the specificity and activity observed, a pNP-C8 binding model for the ester (pNP-C8, N102, and F278 form a flexible bridge) and a specific lipidanchoring mechanism for DAG (L103 and F104 serve as "anchors" to the lipid interface) were proposed. |
| ESTHER : Lan_2015_J.Microbiol.Biotechnol_25_1827 |
| PubMedSearch : Lan_2015_J.Microbiol.Biotechnol_25_1827 |
| PubMedID: 26239010 |
| Gene_locus related to this paper: malgo-a8puy1 |
| Gene_locus related to this paper: malgo-a8puy1 |
Lan D, Wang Q, Popowicz GM, Yang B, Tang Q, Wang Y (2015)
The Role of Residues 103, 104, and 278 in the Activity of SMG1 Lipase from Malassezia globosa: A Site-Directed Mutagenesis Study
J Microbiol Biotechnol
25 :1827
Lan D, Wang Q, Popowicz GM, Yang B, Tang Q, Wang Y (2015)
J Microbiol Biotechnol
25 :1827