Immobilization and characterization of porcine pancreas lipase

Bagi, K.; Simon, Lawrence M.; Szajáni, B.

doi:10.1016/s0141-0229(96)00190-1

Cited by 84 publications

(45 citation statements)

References 17 publications

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“…The standard temperature corresponding to maximum hydrolysis, as well as maximum extent of erucic acid formation, was found to be 35 ºC. This finding is supported by an earlier study (Bagi et al, 1997) where the optimum temperature of porcine pancreas lipase at pH 8.9 was found to be 35 ºC. In the present study, the increase in rate of reaction was higher than the denaturation of lipase at a temperature less than 35 ºC; as a result, the net rate of hydrolysis increased.…”

Section: Effect Of Temperaturesupporting

confidence: 87%

Effects of process variables and additives on mustard oil hydrolysis by porcine pancreas lipase

Goswami

Basu

2012

Braz. J. Chem. Eng.

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-Selective hydrolysis of brown mustard oil (from Brassica juncea) with regioselective porcine pancreas lipase was studied in this work. Buffer and oil phase were considered as the continuous and dispersed phases, respectively. Effects of speed of agitation, pH of the buffer phase, temperature, buffer-oil ratio and enzyme concentration on hydrolysis were observed. The best combination of process variables was: 900 rpm, pH 9, 35 ºC, buffer-oil ratio of 1:1 and enzyme concentration of 10 mg/g oil. These standard conditions led to 50% hydrolysis and selective production of 55% erucic acid in 6 h. Cations like Mg 2+ and Ca 2+ increased hydrolysis, but Cu 2+ strongly inhibited it. Organic solvents decreased hydrolysis, though the decrease was minimum for isooctane. A mixed surfactant comprising of Span 80 and Tween 80 increased erucic acid production by 57% at a buffer-oil ratio of 0.2:1.

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Section: Effect Of Temperaturesupporting

confidence: 87%

Effects of process variables and additives on mustard oil hydrolysis by porcine pancreas lipase

Goswami

Basu

2012

Braz. J. Chem. Eng.

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“…The most popular approaches have been chemical or physical immobilization onto support matrices (17)(18)(19) or physical entrapment in reversed micelles (20,21), which improve the lipase stability and enable the reuse of the biocatalyst. Other approaches used have included chemical or physical modifications of the enzyme (22,23), isolation of natural lipases with stability in organic solvents (24,25), and protein engineering (26,27).…”

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confidence: 99%

Protein Engineering by Random Mutagenesis and Structure-Guided Consensus of Geobacillus stearothermophilus Lipase T6 for Enhanced Stability in Methanol

Dror

Shemesh

Dayan

et al. 2014

Appl Environ Microbiol

116

107

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The abilities of enzymes to catalyze reactions in nonnatural environments of organic solvents have opened new opportunities for enzyme-based industrial processes. However, the main drawback of such processes is that most enzymes have a limited stability in polar organic solvents. In this study, we employed protein engineering methods to generate a lipase for enhanced stability in methanol, which is important for biodiesel production. Two protein engineering approaches, random mutagenesis (error-prone PCR) and structure-guided consensus, were applied in parallel on an unexplored lipase gene from Geobacillus stearothermophilus T6. A high-throughput colorimetric screening assay was used to evaluate lipase activity after an incubation period in high methanol concentrations. Both protein engineering approaches were successful in producing variants with elevated half-life values in 70% methanol. The best variant of the random mutagenesis library, Q185L, exhibited 23-fold-improved stability, yet its methanolysis activity was decreased by one-half compared to the wild type. The best variant from the consensus library, H86Y/ A269T, exhibited 66-fold-improved stability in methanol along with elevated thermostability (؉4.3°C) and a 2-fold-higher fatty acid methyl ester yield from soybean oil. Based on in silico modeling, we suggest that the Q185L substitution facilitates a closed lid conformation that limits access for both the methanol and substrate excess into the active site. The enhanced stability of H86Y/A269T was a result of formation of new hydrogen bonds. These improved characteristics make this variant a potential biocatalyst for biodiesel production.

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“…Covalent immobilization C. rugosa lipase on agarose and silica was also studied, and the retained activity of the immobilized catalyst was found to be between 40% and 80% depending on the support (Moreno et al, 1997). This decrease in activity is generally compensated by an improved stability towards pH and temperature, as well as an improved operational stability allowing recycling of the enzyme (Bagi et al, 1997a). Furthermore due to the membrane thickness and the presence of large numbers of activated groups, it was possible to achieve a maximum enzyme loading of 2.4 mg enzyme/g wet weight of capsules compared to only 0.8 mg/g by adsorption at pH 7.…”

Section: Immobilization By Covalent Linkingmentioning

confidence: 99%

A novel reactive perstraction system based on liquid‐core microcapsules applied to lipase‐catalyzed biotransformations

Wyss¹,

Stockar²,

Marison³

2005

Biotech & Bioengineering

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A novel reactive perstraction system has been developed based on liquid-core capsules, involving an enzyme-catalyzed reaction coupled with simultaneous in situ product recovery. Lipase-catalyzed reactions, hydrolysis of triprionin and nitrophenyl laurate, were selected to test the system and demonstrate the feasibility of immobilization of enzymes to the membranes of liquid-core capsules and the ability to extract hydrophobic products of the reaction within the capsule core. The lipase from Candida rugosa was immobilized to the microcapsules by adsorption and by covalent binding through activation with glutaraldehyde. In both cases improved temperature and operational stability were achieved. Both types of immobilization resulted in a basic shift of the pH optimum for activity, from 7.5 to 9.0. The presence of an organic phase within the capsule core allowed direct product separation and lead to a decrease in product inhibition of the lipase-catalyzed reaction.

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Immobilization and characterization of porcine pancreas lipase

Cited by 84 publications

References 17 publications

Effects of process variables and additives on mustard oil hydrolysis by porcine pancreas lipase

Effects of process variables and additives on mustard oil hydrolysis by porcine pancreas lipase

Protein Engineering by Random Mutagenesis and Structure-Guided Consensus of Geobacillus stearothermophilus Lipase T6 for Enhanced Stability in Methanol

A novel reactive perstraction system based on liquid‐core microcapsules applied to lipase‐catalyzed biotransformations

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