Mutations in the "lid" region affect chain length specificity and thermostability of a Pseudomonas fragi lipase

被引：90

作者：

Santarossa, G ^{[1
]}

Lafranconi, PG ^{[1
]}

Alquati, C ^{[1
]}

DeGioia, L ^{[1
]}

Alberghina, L ^{[1
]}

Fantucci, P ^{[1
]}

Lotti, M ^{[1
]}

机构：

[1] Univ Milan, Dipartimento Biotecnol & Biosci, I-20126 Milan, Italy

来源：

FEBS LETTERS | 2005年 / 579卷 / 11期

关键词：

lipase; chain length selectivity; thermo stability; Pseudomonas fragi;

D O I：

10.1016/j.febslet.2005.03.037

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The cold-adapted Pseudomonas fragi lipase (PFL) displays highest activity on short-chain triglyceride substrates and is rapidly inactivated at moderate temperature. Sequence and structure comparison with homologous lipases endowed with different substrate specificity and stability, pointed to three polar residues in the lid region, that were replaced with the amino acids conserved at equivalent positions in the reference lipases. Substitutions at residues T137 and T138 modified the lipase chain-length preference profile, increasing the relative activity towards C8 substrates. Moreover, mutations conferred to PFL higher temperature stability. On the other hand, replacement of the serine at position 141 by glycine destabilized the protein. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

引用

页码：2383 / 2386

页数：4

共 27 条

[11] ALTERATION OF CHAIN-LENGTH SELECTIVITY OF A RHIZOPUS-DELEMAR LIPASE THROUGH SITE-DIRECTED MUTAGENESIS [J].

JOERGER, RD ;

HAAS, MJ .

LIPIDS, 1994, 29 (06) :377-384

[12] Altered acyl chain length specificity of Rhizopus delemar lipase through mutagenesis and molecular modeling [J].

Klein, RR ;

King, G ;

Moreau, RA ;

Haas, MJ .

LIPIDS, 1997, 32 (02) :123-130

[13] Crystal structure of a bacterial lipase from Chromobacterium viscosum ATCC 6918 refined at 1.6 angstrom resolution [J].

Lang, D ;

Hofmann, B ;

Haalck, L ;

Hecht, HJ ;

Spener, F ;

Schmid, RD ;

Schomburg, D .

JOURNAL OF MOLECULAR BIOLOGY, 1996, 259 (04) :704-717

[14] Structural investigations of the regio- and enantioselectivity of lipases [J].

Lang, DA ;

Dijkstra, BW .

CHEMISTRY AND PHYSICS OF LIPIDS, 1998, 93 (1-2) :115-122

[15] PHOSPHONATE ANALOGS OF TRIACYLGLYCEROLS ARE POTENT INHIBITORS OF LIPASE [J].

MANNESSE, MLM ;

BOOTS, JWP ;

DIJKMAN, R ;

SLOTBOOM, AJ ;

VANDERHIJDEN, HTWV ;

EGMOND, MR ;

VERHEIJ, HM ;

DEHAAS, GH .

BIOCHIMICA ET BIOPHYSICA ACTA-LIPIDS AND LIPID METABOLISM, 1995, 1259 (01) :56-64

[16] The role of Glu87 and Trp89 in the lid of Humicola lanuginosa lipase [J].

Martinelle, M ;

Holmquist, M ;

Clausen, IG ;

Patkar, S ;

Svendsen, A ;

Hult, K .

PROTEIN ENGINEERING, 1996, 9 (06) :519-524

[17] Crystal structure of Pseudomonas aeruginosa lipase in the open conformation -: The prototype for family I.1 of bacterial lipases [J].

Nardini, M ;

Lang, DA ;

Liebeton, K ;

Jaeger, KE ;

Dijkstra, BM .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (40) :31219-31225

[18] THE ALPHA/BETA-HYDROLASE FOLD [J].

OLLIS, DL ;

CHEAH, E ;

CYGLER, M ;

DIJKSTRA, B ;

FROLOW, F ;

FRANKEN, SM ;

HAREL, M ;

REMINGTON, SJ ;

SILMAN, I ;

SCHRAG, J ;

SUSSMAN, JL ;

VERSCHUEREN, KHG ;

GOLDMAN, A .

PROTEIN ENGINEERING, 1992, 5 (03) :197-211

[19] Anatomy of lipase binding sites: the scissile fatty acid binding site [J].

Pleiss, J ;

Fischer, M ;

Schmid, RD .

CHEMISTRY AND PHYSICS OF LIPIDS, 1998, 93 (1-2) :67-80

[20] Lipases as practical biocatalysts [J].

Reetz, MT .

CURRENT OPINION IN CHEMICAL BIOLOGY, 2002, 6 (02) :145-150

← 1 2 3 →