Rational engineering of enzyme stability

Eijsink, Vincent G. H.; Bjørk, Alexandra; Gåseidnes, Sigrid; Sirevåg, Reidun; Synstad, Bjørnar; Burg, Bertus van den; Vriend, Gert

doi:10.1016/j.jbiotec.2004.03.026

Cited by 410 publications

(305 citation statements)

References 141 publications

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“…4 increase ∆G by an entropic destabilization of the unfolded state [18][19][20]. However, particularly introduced disulfide bonds may affect the native state as well since they might prevent unfolding by tethering the native protein structure [21].…”

Section: Accepted M Manuscriptmentioning

confidence: 99%

The effect of additional disulfide bonds on the stability and folding of ribonuclease A

Pecher¹,

Arnold²

2009

Biophysical Chemistry

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International audienceThe significant contribution of disulfide bonds to the conformational stability of proteins is generally considered to result from an entropic destabilization of the unfolded state causing a faster escape of the molecules to the native state. However, the introduction of extra disulfide bonds into proteins as a general approach to protein stabilization yields rather inconsistent results. By modeling studies, we selected positions to introduce additional disulfide bonds into ribonuclease A at regions that had proven to be crucial for the initiation of the folding or unfolding process, respectively. However, only two out of the six variants proved to be more stable than unmodified ribonuclease A. The comparison of the thermodynamic and kinetic data disclosed a more pronounced effect on the unfolding reaction for all variants regardless of the position of the extra disulfide bond. Native-state proteolysis indicated a perturbation of the native state of the destabilized variants that obviously counterbalances the stability gain by the extra disulfide bond

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Section: Accepted M Manuscriptmentioning

confidence: 99%

The effect of additional disulfide bonds on the stability and folding of ribonuclease A

Pecher¹,

Arnold²

2009

Biophysical Chemistry

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“…It is possible that the amino acid sequences comprising the helix D region of plant peroxidases have already maximized thermostabilizing interactions for this region of the protein and that more global stability determinants, not revealed by consensus alignments, occur elsewhere in the molecule. Thermal stability differences between homologous proteins may be due to a very few naturally occurring sequence variations ( [47] and references within). Magliery and Regan [48] have developed a more advanced model which addresses some of the inadequacies of the basic consensus approach [22].…”

Section: Discussionmentioning

confidence: 99%

Consensus mutagenesis reveals that non-helical regions influence thermal stability of horseradish peroxidase

Ryan¹,

O’Connell²,

Ó’Fágáin³

2008

Biochimie

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“…Furthermore, many researchers have attempted to improve enzyme stability and to enhance enzyme activity by rational design and in vitro evolution of enzymes. [19][20][21][22][23] In most cases, however, protein biocatalysts showing the desired properties have not been obtained.…”

Section: Introductionmentioning

confidence: 99%

Enhancement of enzyme activity and stability by poly(γ-glutamic acid)

Lee

Tsujimoto

Uyama

et al. 2010

Polym J

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The effects of poly(c-glutamic acid) (cPGA), a water-soluble poly(amino acid), on the enzyme activity and stability have been investigated. The activity of carbonic anhydrase (CA) was distinctly improved in the presence of 0.30-2.0% cPGA at pH 7. cPGA was also effective to enhance the activity of lipase and a-amylase. cPGA efficiently suppressed denaturation of the enzyme by the thermal treatment and repeated freeze-thaw process. The interaction of cPGA and CA was investigated by using fluorescence probe-labeled protein. The Michaelis-Menten kinetics on the CA-catalyzed hydrolysis of p-nitrophenyl acetate in the absence or presence of cPGA were performed, which demonstrated that the enhancement of the CA activity by cPGA is ascribed to the increase of the catalytic constant k cat .

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Rational engineering of enzyme stability

Cited by 410 publications

References 141 publications

The effect of additional disulfide bonds on the stability and folding of ribonuclease A

The effect of additional disulfide bonds on the stability and folding of ribonuclease A

Consensus mutagenesis reveals that non-helical regions influence thermal stability of horseradish peroxidase

Enhancement of enzyme activity and stability by poly(γ-glutamic acid)

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