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Titolo:
RATIONAL DESIGN OF RHIZOPUS-ORYZAE LIPASE WITH MODIFIED STEREOSELECTIVITY TOWARD TRIRADYLGLYCEROLS
Autore:
SCHEIB H; PLEISS J; STADLER P; KOVAC A; POTTHOFF AP; HAALCK L; SPENER F; PALTAUF F; SCHMID RD;
Indirizzi:
UNIV STUTTGART,INST TECH BIOCHEM,ALLMANDRING 31 D-70569 STUTTGART GERMANY UNIV STUTTGART,INST TECH BIOCHEM D-70569 STUTTGART GERMANY GRAZ TECH UNIV,DEPT BIOCHEM & FOOD CHEM A-8010 GRAZ AUSTRIA INST CHEM & BIOCHEM SENSOR RES D-48149 MUNSTER GERMANY UNIV MUNSTER,DEPT BIOCHEM D-48149 MUNSTER GERMANY
Titolo Testata:
Protein engineering (Print)
fascicolo: 8, volume: 11, anno: 1998,
pagine: 675 - 682
SICI:
0269-2139(1998)11:8<675:RDORLW>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
MICROBIAL LIPASES; PSEUDOMONAS-CEPACIA; HYDROLYSIS; ANALOGS; TRIGLYCERIDES; VALIDATION; MECHANISM; DELEMAR;
Keywords:
MOLECULAR DYNAMICS SIMULATIONS; PROTEIN ENGINEERING; RHIZOPUS ORYZAE LIPASE STEREOSELECTIVITY; TRIRADYLGLYCEROLS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
27
Recensione:
Indirizzi per estratti:
Citazione:
H. Scheib et al., "RATIONAL DESIGN OF RHIZOPUS-ORYZAE LIPASE WITH MODIFIED STEREOSELECTIVITY TOWARD TRIRADYLGLYCEROLS", Protein engineering (Print), 11(8), 1998, pp. 675-682

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) stereo-preference 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 ofROL toward triradylglycerols can be predicted by analyzing the geometry of the substrate docked to ROL: if the torsion angle Phi(O3-C3) Of glycerol is more than 150 degrees, the substrate will preferably be hydrolyzed in sn-l, otherwise in sn-3, For ROL, the following rules werederived: (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 thesize 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.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/11/20 alle ore 10:15:39