Substrate Specificity and Kinetics of <i>Candida rugosa</i> Lipase in Organic Media<sup>a</sup>

Janssen, A.E.M.; Vaidya, A. M.; Halling, Peter J.

doi:10.1111/j.1749-6632.1996.tb33210.x

Cited by 14 publications

(28 citation statements)

References 9 publications

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“…In all the samples tested low specificity was observed with C6. This behavior was previously reported by other authors working with commercial14, 18, 36 and Lip2 purified from natural sources,37 and it has been related to the crystal structures of the CRL isoenzymes Lip1,38, 39 and Lip3 40. They all have a long, narrow, hydrophobic tunnel that can accommodate long acyl‐chain substrates.…”

Section: Resultssupporting

confidence: 75%

Recombinant Candida rugosa lipase 2 from Pichia pastoris: Immobilization and use as biocatalyst in a stereoselective reaction

Benaiges

Alarcón

Fuciños

et al. 2010

Biotechnology Progress

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The characterization of the recombinant Candida rugosa Lip2 (r-Lip2) isoenzyme obtained from fed-batch cultures of Pichia pastoris under PAOX promoter was carried out, determining the optimal pH and temperature as well as their catalytic performance in both hydrolysis and synthesis reactions comparing with purified native Lip2 (n-Lip2) previously determined. The substrate specificity of r-Lip2 in hydrolysis reactions was determined with a series of triacylglycerols and p-nitrophenyl esters of variable acyl chain length. r-Lip2 showed the maximum specificity for both substrates towards medium-chain esters (C-8), similar behavior was observed with n-Lip2. However, significant differences were observed towards unsaturated substrates (triolein) or short-chain esters. A statistical design applied to study the effect of pH and temperature on lipase stability shown that r-Lip2, like n-Lip2, was more sensitive to pH than temperature changes. Nevertheless, the overall stability of soluble r-Lip2 was lower than soluble n-Lip2. The stability of r-lip2 was significantly improved by immobilization onto EP100, an excellent support for lipases with yields around 95% for offered lipolytic activity lower than 600 AU/mL. Finally, immobilized r-Lip2 was tested in the resolution of ibuprofen in isooctane by means of enantioselective esterification using 1-butanol as esterifying agent. r-Lip2 showed a better performance in terms of enantiomeric excess (74%) and enatiomeric factor (96%) than n-Lip2 (56 and 80%, respectively) for the same conversion (40%). Thus, r-Lip2 should be considered a good and pure biocatalyst, easy to produce and with a remaining activity of ca. 90% after one reaction cycle when immobilized on EP100.

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Section: Resultssupporting

confidence: 75%

Recombinant Candida rugosa lipase 2 from Pichia pastoris: Immobilization and use as biocatalyst in a stereoselective reaction

Benaiges

Alarcón

Fuciños

et al. 2010

Biotechnology Progress

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“…This parameter which measures the availability of the species for the biocatalyst, has been recognized as the key parameter for studying enzymes in nonconventional media [4]. It enables an easier comparison of enzyme activity in organic solvent and gaseous environment to be achieved, as substrate-solvent interactions are taken into account [5,6]. Indeed, depending on their polarity, solvents show different ability to solvate the substrates, and this influences the thermodynamic activity of the substrates and thereby the measured enzyme activity [7].…”

Section: Introductionmentioning

confidence: 99%

Water plays a different role on activation thermodynamic parameters of alcoholysis reaction catalyzed by lipase in gaseous and organic media

Graber

Bousquet

Sousa

et al. 2003

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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The effect of water on the alcoholysis of methyl propionate and n-propanol catalyzed by immobilized Candida antarctica lipase B (CALB) has been compared in a continuous solid-gas reactor and in an organic liquid medium. The enthalpic and entropic contributions of water to the Gibbs free energy of activation in the gas phase were different from the ones in the organic phase, the inverse trends being observed for the variation of both DeltaH* and DeltaS* with water activity. Different phenomena were identified for their influence on the thermodynamic parameters. When increasing a(w), the enhanced flexibility of the enzyme was predominant in the gas phase whereas substrate-solvent interactions due to an increased polarity of the solvent affected mainly the thermodynamic parameters in the organic phase. The observed variations of DeltaG* with water activity were in accordance with kinetics results previously obtained in both reaction media.

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“…It is well known that in esterification protonation of carbonyl acid takes place which makes it more electrophilic that undergoes protonation by alcohol forming ester and water as byproduct. This mechanism was also explained by various researches in terms of the kinetic model . First stearic acid binds with enzyme forming acyl enzyme complex which results in the generation of the first product, i.e.…”

Section: Methodsmentioning

confidence: 94%

Acoustic cavitation assisted lipase B catalysed synthesis of polyethylene glycol stearate in a solvent free system via esterification: synthesis and optimization

Nhivekar

Rathod

2019

J of Chemical Tech & Biotech

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BACKGROUND: The current work describes the application of ultrasound and its improving effect on the synthesis of polyethylene glycol (PEG) stearate using Fermase (Fermase CALB™10,000 Candida antarctica Lipase B) as a biocatalyst via esterification of PEG600 and stearic acid. The influence of various parameters like molar ratio, temperature, speed of agitation, enzyme loading, effect of addition of molecular sieves and other factors associated with ultrasound, i.e. duty cycle and power, has been studied and optimized.RESULT: A total of 84.34% of PEG stearate was obtained at ultrasound frequency of 22 kHz with 50 W power, 60 ∘ C temperature, stirring speed 200 rpm, mole ratio of PEG600-stearic acid 1:1, 3% (w/w) molecular sieves, immobilized enzyme loading 0.5% (w/w) and duty cycle 50%. The enzyme showed reusability up to the fifth cycle. The reaction is conducted in a solvent free system which is an additional benefit. CONCLUSION: A maximum conversion of 84.34% was obtained in 3 h of reaction time in the presence of ultrasound as compared to 6 h in the absence of ultrasound denoting the intensified effect of ultrasound on the synthesis.

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Substrate Specificity and Kinetics of Candida rugosa Lipase in Organic Media^a

Cited by 14 publications

References 9 publications

Recombinant Candida rugosa lipase 2 from Pichia pastoris: Immobilization and use as biocatalyst in a stereoselective reaction

Recombinant Candida rugosa lipase 2 from Pichia pastoris: Immobilization and use as biocatalyst in a stereoselective reaction

Water plays a different role on activation thermodynamic parameters of alcoholysis reaction catalyzed by lipase in gaseous and organic media

Acoustic cavitation assisted lipase B catalysed synthesis of polyethylene glycol stearate in a solvent free system via esterification: synthesis and optimization

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Substrate Specificity and Kinetics of Candida rugosa Lipase in Organic Mediaa

Cited by 14 publications

References 9 publications

Recombinant Candida rugosa lipase 2 from Pichia pastoris: Immobilization and use as biocatalyst in a stereoselective reaction

Recombinant Candida rugosa lipase 2 from Pichia pastoris: Immobilization and use as biocatalyst in a stereoselective reaction

Water plays a different role on activation thermodynamic parameters of alcoholysis reaction catalyzed by lipase in gaseous and organic media

Acoustic cavitation assisted lipase B catalysed synthesis of polyethylene glycol stearate in a solvent free system via esterification: synthesis and optimization

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Substrate Specificity and Kinetics of Candida rugosa Lipase in Organic Media^a