Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Scheib, H; Pleiss, J; Stadler, P; Kovac, A; Potthoff, AP; Haalck, L; Spener, F; Paltauf, F; Schmid, RD.
Rational design of Rhizopus oryzae lipase with modified stereoselectivity toward triradylglycerols.
Protein Eng. 1998; 11(8):675-682
Doi: 10.1093/protein/11.8.675
[OPEN ACCESS]
Web of Science
PubMed
FullText
FullText_MUG
Google Scholar
- Co-Autor*innen der Med Uni Graz
- Spener Friedrich
- Altmetrics:
- Dimensions Citations:
- Plum Analytics:
- Scite (citation analytics):
- Abstract:
- The binding site of sn-1(3)-regioselective Rhizopus oryzae lipase (ROL) has been engineered to change the stereoselectivity of hydrolysis of triacylglycerol substrates and analogs. Two types of prochiral triradylglycerols were considered: 'flexible' substrates with ether, benzylether or ester groups, and 'rigid' substrates with amide or phenyl groups, respectively, in the sn-2 position. The molecular basis of sn-1(3) stereoselectivity of ROL was investigated by modeling the interactions between substrates and ROL, and the model was confirmed by experimental determination of the stereoselectivity of wild-type and mutated ROL. For the substrates, the following rules were derived: (i) stereopreference of ROL toward triradylglycerols depends on the substrate structure. Substrates with 'flexible' sn-2 substituents are preferably hydrolyzed at sn-1, 'rigid' substrates at sn-3. (ii) Stereopreference of ROL toward triradylglycerols can be predicted by analyzing the geometry of the substrate docked to ROL: if the torsion angle phiO3-C3 of glycerol is more than 150 degrees, the substrate will preferably be hydrolyzed in sn-1, otherwise in sn-3. For ROL, the following rules were derived: (i) residue 258 affects stereoselectivity by steric interactions with the sn-2 substituent rather than polar interactions. To a lower extent, stereoselectivity is influenced by mutations further apart (L254) from residue 258. (ii) With 'rigid' substrates, increasing the size of the binding site (mutations L258A and L258S) shifts stereoselectivity of hydrolysis toward sn-1, decreasing its size (L258F and L258F/L254F) toward sn-3.
- Find related publications in this database (using NLM MeSH Indexing)
- Glycerol - analogs and derivatives Glycerol - metabolism
- Lipase - genetics Lipase - metabolism
- Models, Molecular -
- Protein Conformation -
- Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism
- Rhizopus - enzymology
- Stereoisomerism -
- Substrate Specificity -
- Find related publications in this database (Keywords)
- molecular dynamics simulations
- protein engineering
- Rhizopus oryzae lipase stereoselectivity
- triradylglycerols