Lipase-catalysed hydrolysis of short-chain substrates in solution and in emulsion: a kinetic study

Nini, Lylia; Sarda, Louis; Comeau, Louis-Claude; Boitard, Elisabeth; Dubès, J.P.; Chahinian, Henri

doi:10.1016/s1388-1981(01)00172-x

Cited by 50 publications

(37 citation statements)

References 47 publications

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“…The major component of the cuticle is cutin, an insoluble polyester composed mainly of saturated C 16 and C 18 hydroxy and epoxy fatty acids (14,16,27,38). During the process of scouring in the textile industry, the cuticle layer has to be removed in order to improve the wettability of cotton fiber, which then facilitates uniform dyeing and finishing.…”

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confidence: 99%

Characterization of Thermobifida fusca Cutinase-Carbohydrate-Binding Module Fusion Proteins and Their Potential Application in Bioscouring

Zhang

Chen

et al. 2010

Appl Environ Microbiol

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Cutinase from Thermobifida fusca is thermally stable and has potential application in the bioscouring of cotton in the textile industry. In the present study, the carbohydrate-binding modules (CBMs) from T. fusca cellulase Cel6A (CBM Cel6A ) and Cellulomonas fimi cellulase CenA (CBM CenA ) were fused, separately, to the carboxyl terminus of T. fusca cutinase. Both fusion enzymes, cutinase-CBM Cel6A and cutinase-CBM CenA , were expressed in Escherichia coli and purified to homogeneity. Enzyme characterization showed that both displayed similar catalytic properties and pH stabilities in response to T. fusca cutinase. In addition, both fusion proteins displayed an activity half-life of 53 h at their optimal temperature of 50°C. Compared to T. fusca cutinase, in the absence of pectinase, the binding activity on cotton fiber was enhanced by 2% for cutinase-CBM Cel6A and by 28% for cutinase-CBM CenA , whereas in the presence of pectinase, the binding activity was enhanced by 40% for the former and 45% for the latter. Notably, a dramatic increase of up to 3-fold was observed in the amount of released fatty acids from cotton fiber by both cutinase-CBM fusion proteins when acting in concert with pectinase. This is the first report of improving the scouring efficiency of cutinase by fusing it with CBM. The improvement in activity and the strong synergistic effect between the fusion proteins and pectinase suggest that they may have better applications in textile bioscouring than the native cutinase.Cotton fiber has a multilayered structure, with its outermost surface being the cuticle that is cross-linked to the primary cell wall of cotton fiber by esterified pectin substances. The major component of the cuticle is cutin, an insoluble polyester composed mainly of saturated C 16 and C 18 hydroxy and epoxy fatty acids (14,16,27,38). During the process of scouring in the textile industry, the cuticle layer has to be removed in order to improve the wettability of cotton fiber, which then facilitates uniform dyeing and finishing. Traditionally, this process is performed by hot hydrolysis in alkaline medium, which not only consumes large quantities of water and energy but also causes severe pollution and fiber damage (20,21,33). Therefore, environment-friendly scouring methods based on biocatalysts have been actively sought (2,30,36).Cutinase is a multifunctional esterase capable of degrading the cutin component of the cuticle. Earlier reports showed that the fungal cutinase from Fusarium solani pisi has potential use for cotton cuticle degradation and exhibits a good synergistic effect with pectinase, an enzyme utilized to degrade pectin, in the scouring of cotton fiber (1,7,8,14). Moreover, site-directed mutagenesis has been performed to replace the specific amino acid residues near the active site of cutinase (3) to improve its hydrolytic activity toward polyesters. More recently, a cutinase from the thermophilic bacterium Thermobifida fusca has been identified and overexpressed in Escherichia coli in our laboratory (10). The...

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confidence: 99%

Characterization of Thermobifida fusca Cutinase-Carbohydrate-Binding Module Fusion Proteins and Their Potential Application in Bioscouring

Zhang

Chen

et al. 2010

Appl Environ Microbiol

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“…It rules out the idea of a strict correlation between the interfacial activation process and the presence of a lid. However, it must be kept in mind that some authors emphasize the ambiguity of the phenomenon of interfacial activation (33,34). We have looked for tilted peptides in the lid sequences of several PLRP2 displaying or not an interfacial activation process.…”

Section: Kinetic Parameters Of Native and Mutant Lipases On Tributyrinmentioning

confidence: 99%

Role of the Lid Hydrophobicity Pattern in Pancreatic Lipase Activity

Thomas

Allouche

Basyn

et al. 2005

Journal of Biological Chemistry

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Pancreatic lipase is a soluble globular protein that must undergo structural modifications before it can hydrolyze oil droplets coated with bile salts. The binding of colipase and movement of the lipase lid open access to the active site. Mechanisms triggering lid mobility are unclear. The *KNILSQIVDIDGI* fragment of the lid of the human pancreatic lipase is predicted by molecular modeling to be a tilted peptide. Tilted peptides are hydrophobicity motifs involved in membrane fusion and more globally in perturbations of hydrophobic/hydrophilic interfaces. Analysis of this lid fragment predicts no clear consensus of secondary structure that suggests that its structure is not strongly sequence determined and could vary with environment. Point mutations were designed to modify the hydrophobicity profile of the [240 -252] fragment and their consequences on the lipase-mediated catalysis were tested. Two mutants, in which the tilted peptide motif was lost, also have poor activity on bile saltcoated oil droplets and cannot be reactivated by colipase. Conversely, one mutant in which a different tilted peptide is created retains colipase dependence. These results suggest that the tilted hydrophobicity pattern of the [240 -252] fragment is neither important for colipase binding to lipase, nor for interfacial binding but is important to trigger the maximal catalytic efficiency of lipase in the presence of bile salt.The pancreatic lipase-colipase complex plays a key role in dietary fat absorption in the intestine by converting triglycerides into more polar products able to cross the brush-border membrane of enterocytes. The lipase is fully active on water-insoluble substrates that form lipid/water interfaces, a phenomenon called interfacial activation. In vivo, oil droplets consist of a bulk substrate phase surrounded by a monolayer of amphiphilic compounds, mainly biliary lipids (phosphatidylcholine, cholesterol and bile salts) that prevent lipase adsorption. To counteract the inhibitory effect of biliary lipids, the pancreas secretes a small protein, colipase, which anchors lipase to the biliary lipid-coated water/ lipid interface. The water-soluble lipase must therefore partition between aqueous and lipid phases before lipolysis can occur.Structural studies (1-3) have shown that lipases possess a two-domain organization, the N-terminal domain bearing the active site and the C-terminal domain bearing the colipase binding site. One specific feature of lipase is the shielding of its catalytic site by a surface loop (lid) controlling the access of the substrate. Therefore, movement of the lid domain is an absolute requirement for the lipase to adopt an active conformation.The role of the lid in lipolysis has been investigated by site-directed mutagenesis, lid exchange, or lid deletion (4 -6). It was first thought to account for the interfacial activation of pancreatic lipase but the presence of a full-length lid in most pancreatic lipase-related proteins that display no interfacial activation rules out this explanation. Actua...

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“…They can also catalyze the reverse esterification reactions, thus producing glycerides from glycerol and fatty acids. Many lipases are also involved in the catalysis of trans-esterification reactions and enantioselective hydrolyses 39,40,41,42 . Most commercial lipases are obtained from fungi (mainly Rhizopus, Candida, and Rhizomucor) and bacteria (Pseudomonas and Chromobacterium) 43 .…”

Section: Lipasementioning

confidence: 99%

Industrial Important Enzymes from Alkaliphiles – An overview

Chinnathambi¹

2015

Biosci., Biotech. Res. Asia

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Alkaliphiles are an interesting group of extremophilic organisms that thrive at pH of 9.0 and above and which are represented by prokaryotes, eukaryotes, and archaea. It is clear that a variety of taxa are represented among the alkaliphiles, some of which are new. Alkaliphiles can be isolated from normal environments such as garden soil, although the number isolated increases dramatically in highly alkaline environments. Many alkaliphile products, notably enzymes, have found widespread practical applications, primarily in the detergent and laundry industries; other potentially important products isolated from alkaliphiles include antibiotics and carotenoids. Alkaliphiles, and their products, are also potentially important for use in the degradation of xenobiotics, and they play a major role in the biogeocycling of key inorganic compounds. This review provides an insight into the huge diversity of alkaliphilic bacteria, the varied products obtained from them, and the need for further investigations on this interesting group of bacteria.

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Lipase-catalysed hydrolysis of short-chain substrates in solution and in emulsion: a kinetic study

Cited by 50 publications

References 47 publications

Characterization of Thermobifida fusca Cutinase-Carbohydrate-Binding Module Fusion Proteins and Their Potential Application in Bioscouring

Characterization of Thermobifida fusca Cutinase-Carbohydrate-Binding Module Fusion Proteins and Their Potential Application in Bioscouring

Role of the Lid Hydrophobicity Pattern in Pancreatic Lipase Activity

Industrial Important Enzymes from Alkaliphiles – An overview

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