Directed enzyme evolution: beyond the low-hanging fruit

被引：116

作者：

Goldsmith, Moshe ^{[1
]}

Tawfik, Dan S. ^{[1
]}

机构：

[1] Weizmann Inst Sci, Dept Biol Chem, IL-76100 Rehovot, Israel

来源：

CURRENT OPINION IN STRUCTURAL BIOLOGY | 2012年 / 22卷 / 04期

基金：

以色列科学基金会; 美国国家卫生研究院;

关键词：

IN-VITRO; LABORATORY EVOLUTION; DESIGNED LIBRARIES; RANDOM MUTAGENESIS; PROTEIN; DISPLAY; SPECIFICITY; DIVERSITY; SELECTION; STEREOSELECTIVITY;

D O I：

10.1016/j.sbi.2012.03.010

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 field of directed evolution has progressed to the point where it is feasible to engineer enzymes for unnatural substrates and reactions with catalytic efficiencies and regio-specificity or stereo-specificity that rival those of natural enzymes. Here, we describe the conceptual and methodological advances that have enabled this progress. We address methodologies based on small libraries enriched with improved variants and carrying compensatory stabilizing mutations. Such libraries can be combined with low-throughput screens that provide high accuracy and directly target the desired substrate and reaction conditions, and thereby provide highly improved variants.

引用

收藏

页码：406 / 412

页数：7

相关论文

共 102 条

[1] Ultrahigh-throughput screening in drop-based microfluidics for directed evolution [J].

Agresti, Jeremy J. ;

Antipov, Eugene ;

Abate, Adam R. ;

Ahn, Keunho ;

Rowat, Amy C. ;

Baret, Jean-Christophe ;

Marquez, Manuel ;

Klibanov, Alexander M. ;

Griffiths, Andrew D. ;

Weitz, David A. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2010, 107 (09) :4004-4009

[2] Directed Evolution of Sulfotransferases and Paraoxonases by Ancestral Libraries [J].

Alcolombri, Uria ;

Elias, Mikael ;

Tawfik, Dan S. .

JOURNAL OF MOLECULAR BIOLOGY, 2011, 411 (04) :837-853

[3] The Transition of Human Estrogen Sulfotransferase from Generalist to Specialist Using Directed Enzyme Evolution [J].

Amar, Dotan ;

Berger, Ilana ;

Amara, Neri ;

Tafa, Gemechu ;

Meijler, Michael M. ;

Aharoni, Amir .

JOURNAL OF MOLECULAR BIOLOGY, 2012, 416 (01) :21-32

[4] Developing recombinant antibodies for biomarker detection [J].

Baird, Cheryl L. ;

Fischer, Christopher J. ;

Pefaur, Noah B. ;

Miller, Keith D. ;

Kagan, Jacob ;

Srivastava, Sudhir ;

Rodland, Karin D. .

CANCER BIOMARKERS, 2010, 6 (5-6) :271-279

[5] An exciting but challenging road ahead for computational enzyme design [J].

Baker, David .

PROTEIN SCIENCE, 2010, 19 (10) :1817-1819

[6] The Moderately Efficient Enzyme: Evolutionary and Physicochemical Trends Shaping Enzyme Parameters [J].

Bar-Even, Arren ;

Noor, Elad ;

Savir, Yonatan ;

Liebermeister, Wolfram ;

Davidi, Dan ;

Tawfik, Dan S. ;

Milo, Ron .

BIOCHEMISTRY, 2011, 50 (21) :4402-4410

[7] Discovery and Protein Engineering of Biocatalysts for Organic Synthesis [J].

Behrens, Geoffrey A. ;

Hummel, Anke ;

Padhi, Santosh K. ;

Schaetzle, Sebastian ;

Bornscheuer, Uwe T. .

ADVANCED SYNTHESIS & CATALYSIS, 2011, 353 (13) :2191-2215

[8] The Molecular Basis for the Broad Substrate Specificity of Human Sulfotransferase 1A1 [J].

Berger, Ilana ;

Guttman, Chen ;

Amar, Dotan ;

Zarivach, Raz ;

Aharoni, Amir .

PLOS ONE, 2011, 6 (11)

[9] Ohno's Model Revisited: Measuring the Frequency of Potentially Adaptive Mutations under Various Mutational Drifts [J].

Bershtein, Shimon ;

Tawfik, Dan S. .

MOLECULAR BIOLOGY AND EVOLUTION, 2008, 25 (11) :2311-2318

[10] Intense neutral drifts yield robust and evolvable consensus proteins [J].

Bershtein, Shimon ;

Goldin, Korina ;

Tawfik, Dan S. .

JOURNAL OF MOLECULAR BIOLOGY, 2008, 379 (05) :1029-1044

← 1 2 3 4 5 6 7 8 9 10 →