Effect of mutations in Candida antarctica B lipase

Patkar, Shamkant; Vind, Jesper; Kelstrup, Ebbe; Christensen, Morten Würtz; Svendsen, Allan; Borch, Kim; Kirk, Ole

doi:10.1016/s0009-3084(98)00032-2

Cited by 59 publications

(39 citation statements)

References 13 publications

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“…We have previously reported thermostability, activity, and enantioselectivity for a few single point mutated CALB variants. [13,14] More recently, other groups have also generated and reported CALB variants with improved properties. [15] A group from Schering-Plough used error-prone PCR to produce mutants with increased thermostability [16] and DNA family shuffling to produce other mutants with increased hydrolysis activity toward a prochiral diester substrate.…”

Section: Introductionmentioning

confidence: 99%

Understanding the Plasticity of the α/β Hydrolase Fold: Lid Swapping on the Candida antarctica Lipase B Results in Chimeras with Interesting Biocatalytic Properties

et al. 2009

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The best of both worlds. Long molecular dynamics (MD) simulations of Candida antarctica lipase B (CALB) confirmed the function of helix alpha5 as a lid structure. Replacement of the helix with corresponding lid regions from CALB homologues from Neurospora crassa and Gibberella zeae resulted in new CALB chimeras with novel biocatalytic properties. The figure shows a snapshot from the MD simulation. The Candida antarctica lipase B (CALB) has found very extensive use in biocatalysis reactions. Long molecular dynamics simulations of CALB in explicit aqueous solvent confirmed the high mobility of the regions lining the channel that leads into the active site, in particular, of helices alpha5 and alpha10. The simulation also confirmed the function of helix alpha5 as a lid of the lipase. Replacing it with corresponding lid regions from the CALB homologues from Neurospora crassa and Gibberella zeae resulted in two new CALB mutants. Characterization of these revealed several interesting properties, including increased hydrolytic activity on simple esters, specifically substrates with C(alpha) branching on the carboxylic side, and much increased enantioselectivity in hydrolysis of racemic ethyl 2-phenylpropanoate (E>50), which is a common structure of the profen drug family.

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Section: Introductionmentioning

confidence: 99%

Understanding the Plasticity of the α/β Hydrolase Fold: Lid Swapping on the Candida antarctica Lipase B Results in Chimeras with Interesting Biocatalytic Properties

et al. 2009

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“…In some cases, simultaneous improvement of thermostability and enzyme activity were obtained [Khurana et al, 2011;Kolling et al, 2010], while findings to the contrary have been reported in other studies [Patkar et al, 1998;Sharma et al, 2014]. Here, substitution of structural Lys173 with Ala increased both activity and thermostability of the lipase variant.…”

Section: Biochemical Characteristicsmentioning

confidence: 62%

“…NTU 03 had reduced specific activity. Similarly, a T103G mutation in Candida antarctica B lipase increased thermal stability of the lipase and reduced its specific activity [Patkar et al, 1998]. …”

Section: Introductionmentioning

confidence: 99%

Rational Design of K173A Substitution Enhances Thermostability Coupled with Catalytic Activity of <b><i>Enterobacter</i></b> sp. Bn12 Lipase

2014

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ELBn12 is a lipase isolated from Enterobacter sp. Bn12 with potential application in biotechnology. Homology modeling and rational design were applied to improve thermal stability of the lipase. K173A substitution introduced an AXXXA motif on the lipase model and it may have role in dimerization and thermostability of the protein. Site-directed mutagenesis was performed to construct the lipase variant. The mutated lipase was expressed in Escherichia coli pLysS and partially purified. Thermostability of the mutated lipase after 1 h of incubation at 70°C was twice that of wild-type lipase under the same conditions. Catalytic activity of the variant was about 1.5-fold towards tricaprylin at 60°C and pH 8.0; moreover, the lipase variant preserved its stability within the pH range of 7.0-11.0. Substitution of superficial hydrophilic Lys with hydrophobic Ala residue increased stability of the mutated lipase in the presence of nonionic surfactants, but this substitution caused lower stability towards polar solvents. Analysis of circular dichroism spectroscopy showed that the K173A mutation altered the secondary structure of the lipase into a more helical one. In conclusion, results of this study demonstrate the positive role of generation of a stabilizing protein motif through rational protein engineering that improves the enzyme characteristics.

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“…[93] Some reports determined that the optimal range for carrying out these reactions was in the range of 70-90 8C. [94][95][96] Frampton et al [97] reported the synthesis of siloxane-containing polyesters and polyamides by using immobilized CalB at temperatures that werea bove the denaturation point of the CalB molecule.…”

Section: Thermal Tolerance Of Calb In the Presence Of Siloxanesmentioning

confidence: 99%

Biocatalysis in Silicon Chemistry

Frampton

Zelisko

2017

Chemistry An Asian Journal

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The application of biocatalytic or chemoenzymatic techniquesi ns ilicon chemistry serves two roles:i tp rovides ag reater understanding of the processing of silicon species by natural systems, such as plants, diatoms, and sponges, as well openingu pa venues to green methodologies in the field. In the latter case, biocatalytic approaches have been applied to the synthesis of small-molecule systems and polymeric materials. Often these biocatalytic approaches allow access to molecular structures under mild conditions and, in some cases, molecular structures that are not accessible throught raditional chemical methodologies. Ar eview of recent advances in the applications of biocatalysis in silicon chemistry is presented.

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Effect of mutations in Candida antarctica B lipase

Cited by 59 publications

References 13 publications

Understanding the Plasticity of the α/β Hydrolase Fold: Lid Swapping on the Candida antarctica Lipase B Results in Chimeras with Interesting Biocatalytic Properties

Understanding the Plasticity of the α/β Hydrolase Fold: Lid Swapping on the Candida antarctica Lipase B Results in Chimeras with Interesting Biocatalytic Properties

Rational Design of K173A Substitution Enhances Thermostability Coupled with Catalytic Activity of <b><i>Enterobacter</i></b> sp. Bn12 Lipase

Biocatalysis in Silicon Chemistry

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