Directed evolution of enzymes and pathways for industrial biocatalysis

Zhao, Huimin; Chockalingam, Karuppiah; Chen, Zhilei

doi:10.1016/s0958-1669(02)00291-4

Cited by 155 publications

(83 citation statements)

References 63 publications

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“…In vitro-directed evolution through DNA shuffling has been routinely applied to evolve enzymes for basic research or to improve proteins of industrial significance, such as improved enzyme kinetics (16,18,41,42,50) and altered substrate or product specificities (17,28,33,48), as well as vaccine and pharmaceutical development (26,43). In the case of directed evolution targeting the glyphosate resistance of EPSPS, both He et al and Zhou et al also obtained mutant EPSPS with increased glyphosate resistance and enzyme activity (13,50).…”

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confidence: 99%

Improvement of Glyphosate Resistance through Concurrent Mutations in Three Amino Acids of the Ochrobactrum 5-Enopyruvylshikimate-3-Phosphate Synthase

Tian

Xiong

et al. 2011

Appl Environ Microbiol

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A mutant of 5-enopyruvylshikimate-3-phosphate synthase from Ochrobactrum anthropi was identified after four rounds of DNA shuffling and screening. Its ability to restore the growth of the mutant ER2799 cell on an M9 minimal medium containing 300 mM glyphosate led to its identification. The mutant had mutations in seven amino acids: E145G, N163H, N267S, P318R, M377V, M425T, and P438L. Among these mutations, N267S, P318R, and M425T have never been previously reported as important residues for glyphosate resistance. However, in the present study they were found by site-directed mutagenesis to collectively contribute to the improvement of glyphosate tolerance. Kinetic analyses of these three mutants demonstrated that the effectiveness of these three individual amino acid alterations on glyphosate tolerance was in the order P318R > M425T > N267S. The results of the kinetic analyses combined with a three-dimensional structure modeling of the location of P318R and M425T demonstrate that the lower hemisphere's upper surface is possibly another important region for glyphosate resistance. Furthermore, the transgenic Arabidopsis was obtained to confirm the potential of the mutant in developing glyphosate-resistant crops.

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confidence: 99%

Improvement of Glyphosate Resistance through Concurrent Mutations in Three Amino Acids of the Ochrobactrum 5-Enopyruvylshikimate-3-Phosphate Synthase

Tian

Xiong

et al. 2011

Appl Environ Microbiol

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“…Directed evolution approaches are frequently used to explore, manipulate, and optimize enzyme properties, including substrate specificity, catalytic activity, thermostability, and solubility, because no information about the tertiary structure of the enzyme is required [90][91][92][93]. These approaches are also applicable to the analysis of mutation effects on enzyme activity to provide useful information for the improvement of enzyme function.…”

Section: Protein Engineering Of the Sbd Region Of Phaz Rpit1mentioning

confidence: 99%

Biodegradable Polymers: Recent Developments and New Perspectives

Carrasco¹,

Pérez²,

Cailloux³

et al. 2017

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“…Methods for engineering an enzyme for industrial use typically include directed evolution, rational design or a combination of the two 15 . Directed evolution mimics the process of evolution by introducing mutations and selecting variants with the desired function by application of a selective pressure.…”

Section: Applicationmentioning

confidence: 99%

Cofactor binding determinants in the NADPH-dependent nitrile-oxidoreductase QueF

Cohen¹

2016

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The NADPH-dependent nitrile-oxidioreductase, QueF is the only known enzyme capable of reducing a nitrile group to an amine. This ability makes it an attractive alternative to conventional industrial nitrile redution. Understating how QueF binds NADPH may lead to the development of an enzyme capable of using the less expensive reductive cofactor NADH. A cofactor docking model indicates that key residues, Q60 and Y21, interact with the ribose phosphate moiety unique to NADPH. Mutants of these residues (Q60A, Q60N, Q60E, Y21A and Y21F) were developed for steady-state kinetic analysis. Modification to either residue resulted in a decrease in binding and catalytic activity when using NADPH as a hydride source. Q60E had the greatest reduction in activity (0.45% of WT). There appears to be both charge and steric factors involved in direct cofactor binding. Neither WT or mutant enzymes were able to utilize NADH as a reductive cofactor to any observable extent.

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Directed evolution of enzymes and pathways for industrial biocatalysis

Cited by 155 publications

References 63 publications

Improvement of Glyphosate Resistance through Concurrent Mutations in Three Amino Acids of the Ochrobactrum 5-Enopyruvylshikimate-3-Phosphate Synthase

Improvement of Glyphosate Resistance through Concurrent Mutations in Three Amino Acids of the Ochrobactrum 5-Enopyruvylshikimate-3-Phosphate Synthase

Biodegradable Polymers: Recent Developments and New Perspectives

Cofactor binding determinants in the NADPH-dependent nitrile-oxidoreductase QueF

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