Improving esterification activity of Burkholderia cepacia lipase encapsulated in silica by bioimprinting with substrate analogues

Cao, Xiongwen; Yang, Jiangke; Shu, Lei; Yu, Benqin; Yan, Yunjun

doi:10.1016/j.procbio.2008.10.003

Cited by 40 publications

(32 citation statements)

References 23 publications

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“…[32] However, there are only a few publications on the mutual effects of sol-gel entrapment and bioimprinting for lipases. [27,33] Imprinting effects of ad hoc selected molecules involving lauric acid (LA), [33] Tween 80 [27] and other molecules (18-crown-6, [27] methyl-b-cyclodextrin [27] ) were investigated so far. Our goal is to investigate the imprinting effect of rationally selected molecules (compounds which were found as substrate mimics in experimental X-ray structures of lipases and their structural analogues) in sol-gel process resulting in robust immobilized biocatalysts for continuous-flow kinetic resolutions.…”

Section: Introductionmentioning

confidence: 99%

Novel Sol‐Gel Lipases by Designed Bioimprinting for Continuous‐Flow Kinetic Resolutions

et al. 2011

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Abstract:The bioimprinting effect in sol-gel immobilization of lipases was studied to develop efficient novel immobilized biocatalysts with significantly improved properties for biotransformations in continuous-flow systems. The bioimprinting candidates were selected systematically among the substrate mimics already found in the active site of experimental lipase structures. Four lipases (Lipase AK, Lipase PS, CaLB and CrL) were immobilized by a sol-gel process with nine bioimprinting candidates using various combinations of tetraethoxysilane (TEOS), phenyltriethoxysilane (PhTEOS), octyltriethoxysilane (OcTEOS) and dimethyldiethylsilane (DMDEOS) as silica precursors. The biocatalytic properties of the immobilized lipases were characterized by enantiomer selective acylation of various racemic secondary alcohols in two different multisubstrate systems (mixture A: a series of alkan-2-ols rac-1a-e and mixture B: heptan-2-ol rac-1f and 1-phenylethanol rac1g). Except with Lipase AK, the most significant activity enhancement was found with the imprinting molecules already found as substrate mimics in Xray structures of various lipases. The synthetic usefulness of the best biocatalysts was demonstrated by the kinetic resolution of racemic 1-(thiophen-2-yl)ethanol (rac-1h) in batch and continuous-flow systems.

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Section: Introductionmentioning

confidence: 99%

Novel Sol‐Gel Lipases by Designed Bioimprinting for Continuous‐Flow Kinetic Resolutions

et al. 2011

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“…Thus, the substrate of the reaction served as the best imprint molecule. Similarly, bioimprinting of CRL with OA activated the lipase in esterification of OA and ethanol (Foresti et al 2005) and bioimprinting of Burkholderia cepacia lipase with lauric acid resulted in activation factor of 40 in esterification of lauric acid and lauryl alcohol in isooctane (Cao et al 2009). Yan et al (2009) reported that, when OA was used as for bioimprinting of Geotrichum candidum lipase, the activation achieved in methyl oleate synthesis was due to a combination of interfacial activation and conformational changes in the active site of the enzyme which matches the substrate.…”

Section: Resultsmentioning

confidence: 99%

Enhancement of activity and selectivity of Candida rugosa lipase and Candida antarctica lipase A by bioimprinting and/or immobilization for application in the selective ethanolysis of fish oil

Kahveci

2011

Biotechnol Lett

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Candida rugosa lipase (CRL) and Candida antarctica lipase A (CALA) with improved activity and selectivity were prepared for use in organic solvent media. CRL bioimprinted with fatty acids exhibited eightfold enhanced transesterification activity in hexane. Combination of bioimprinting and coating with lecithin or with immobilization did not improve the activity further. CALA was immobilized with and without bioimprinting, none of which improved the activity. All modified lipases were tested for selective ethanolysis of fish oil to concentrate omega-3 polyunsaturated fatty acids (PUFA). None of the preparations, except the immobilized ones catalysed ethanolysis. Immobilized CRL-catalyzed ethanolysis giving 27% (v/v) ethyl esters (EE) in 48 h, of which 43 mol% was oleic acid but no PUFA was detected in the EE fraction. Fatty acid selectivity of CALA was significantly improved by immobilization combined with bioimprinting, resulting in 5.5-fold lower omega-3 PUFA in EE.

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“…The enzyme is already proven to have versatile application [26][27][28][29] and its crystal structure 30 has been resolved earlier. Like other lipases it poses a catalytic triad of Ser-His-Asp buried deep in to an oxyanion hole (Ser has been indicated by yellow arrow in Fig.…”

Section: Structural Analysis Of Lipasementioning

confidence: 99%

Facile fabrication of lipase to amine functionalized gold nanoparticles to enhance stability and activity

2017

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Among various techniques of immobilization, EDC/NHS cross linking is a simple and single step process for covalent coupling between enzymes and nanoparticles. Here we describe immobilization of lipase on amine functionalized gold nanoparticles (AuNPs-NH 2 ) to attain enhanced activity and stability. To achieve a suitable orientation, it is necessary to understand the contribution of different functional groups on the enzyme's surface. Therefore, the crystal structure of lipase was analyzed using a computational method (PyMOL) to find the exposed acidic amino acid residues that can be exploited for conjugation. Confirmation of conjugation (AuNP-NH 2 -lipase) was determined by various techniques such as agarose gel electrophoresis, zeta measurement, FTIR-spectroscopy and TEM. Further, catalytic parameters (V max , K M,app , K cat , and K cat /K M,app ) have been studied to establish activity enhancement upon immobilization. The data also suggested that, AuNP-NH 2 -lipase has desirable improved parameters such as temperature and storage stability. The thermodynamic parameters for the kinetics of deactivation IntroductionEnzymes have wide application in food, detergent, medicine, diagnostics, energy and many other industrial sectors due to their catalytic activity under mild conditions, specicity, costeffectiveness and greener production conditions. 1,2 However industrial application of free enzymes is restricted due to high lability, poor stability and problems in their recovery and reuse. 3Immobilization is the effective way to overcome these limitations and extend the horizon of their applications.4 Various methods of immobilization include physical adsorption, ionic interaction, covalent bonding between support and enzyme or entrapment within the different matrices.5 Hydrophobic interactions, ionic interactions, hydrogen bonds, van der Waals interactions and solvation are the major forces responsible for non-covalent interactions. Except for covalent attachment, the above mentioned techniques mainly rely on weak interactions, therefore enzymes are prone to detachment from the support surface with time. Covalent bonding mediated immobilization helps in improving enzyme activity at the cost of loss of conformational integrity.6 A very simple method of crosslinking biomolecules to nanoparticles relies on EDC/NHS coupling in which carbodiimides mediate formation of amide bond between carboxy and amine groups.7 High solubility in water, ease to remove byproducts, zero length cross linking molecule and single step reaction provides an edge over other methods in EDC/NHS coupling.8 Several enzymes have been conjugated using this strategy, but they lack in their structure based consideration prior to immobilization. Improper orientation aer immobilization ultimately affects activity; therefore, it is highly essential to understand the structure of protein before immobilization. One of the rationale approaches is to exploit specic surface amino acids for conjugating enzymes with the nanosurface so that the catalytic p...

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Improving esterification activity of Burkholderia cepacia lipase encapsulated in silica by bioimprinting with substrate analogues

Cited by 40 publications

References 23 publications

Novel Sol‐Gel Lipases by Designed Bioimprinting for Continuous‐Flow Kinetic Resolutions

Novel Sol‐Gel Lipases by Designed Bioimprinting for Continuous‐Flow Kinetic Resolutions

Enhancement of activity and selectivity of Candida rugosa lipase and Candida antarctica lipase A by bioimprinting and/or immobilization for application in the selective ethanolysis of fish oil

Facile fabrication of lipase to amine functionalized gold nanoparticles to enhance stability and activity

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