Esterase Active in Polar Organic Solvents from the Yeast<i>Pseudozyma</i>sp. NII 08165

Alex, Deepthy; Shainu, Anju; Pandey, Ashok; Sukumaran, Rajeev K.

doi:10.1155/2014/494682

Cited by 13 publications

(10 citation statements)

References 26 publications

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“…Generally, esterases from yeasts (including Aureobasidium spp.) present optimal pH and temperature in the range 7.0-8.0 and 35-60°C, respectively [36,57,59,61]. Thus, the optimal temperature found for esterase from A. pullulans is in agreement with the literature.…”

Section: Tablesupporting

confidence: 89%

“…Tolerance to more polar solvents is considered a desired property for esterases involved in some asymmetric synthesis since these water miscible solvents can act as a homogeneous co-solvent and enhance the solubility of both the organic substrates and products [61,71]. Therefore, an esterase which preserves or improves its activity in polar solvents assumes special relevance in this context.…”

Section: Effect Of Solventsmentioning

confidence: 99%

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Integrated strategy for purification of esterase from Aureobasidium pullulans

Lemes

Silvério

Rodrigues

et al. 2019

Separation and Purification Technology

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Esterases catalyze the cleavage and formation of ester bonds of a broad range of substrates presenting a widespread spectrum of industrial applications. This work aimed to partially purify and characterize an esterase from Aureobasidium pullulans LABIOTEC 01 produced in a culture medium containing olive mill wastewater. Esterase purification was evaluated using different strategies, namely the enzyme recovery by PEG-salt aqueous twophase systems (ATPSs); and the enzyme precipitation with ammonium sulfate, acetone, and ethanol. The best purification factor (18 ± 2) was obtained when the ATPS composed of 20% (w/w) polyethylene glycol (PEG) 6000 and 5.8% (w/w) potassium phosphate buffer (PPB) pH 8.0 was combined with acetone precipitation. The partially purified enzyme presented an optimum pH of 5.0, although it remained active in the pH range of 4.5 to 7.5 (≥ 50% relative activity). The optimum temperature was found to be 60°C. Furthermore, the addition of salts such as FeCl 3 , CuSO 4 and MnCl 2 promoted an increase in the enzymatic activity (above 100%). The enzyme was found to be stable and showed high activity when exposed to polar solvents such as dimethyl sulfoxide, dimethylformamide, and methanol. The use of an integrated strategy of purification combining simple purification methods such as ATPS and precipitation was herein reported for the first time for esterase. This strategy proved to be an interesting approach to partially purify the esterase produced under submerged fermentation by A. pullulans. Furthermore, the enzyme showed potential to be applied in industrial biocatalytic processes using high temperature and different salts or solvents. Also, the production of esterase using olive mill wastewater as substrate demonstrated to be a suitable alternative to reduce and valorize agro-industrial residues.

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Section: Tablesupporting

confidence: 89%

Section: Effect Of Solventsmentioning

confidence: 99%

Integrated strategy for purification of esterase from Aureobasidium pullulans

Lemes

Silvério

Rodrigues

et al. 2019

Separation and Purification Technology

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“…The tolerance of PmEst to water miscible organic solvents is quite significant and could be related to the Pseudozyma sp . NII 08165 esterase [ 15 ] that preserved 80% of activity in presence of 25% ethanol. On the other hand, other examples of thermostable esterases isolated from Thermotoga maritima [ 43 ] and Salimicrobium [ 12 ] showed a dramatic loss of activity (reduction of ~80%) in the presence of 10% ethanol and propanol.…”

Section: Resultsmentioning

confidence: 99%

“…The use of enzymes in organic solvents offers numerous potential advantages compared to traditional aqueous enzymology, such as higher solubility of hydrophobic substrates, reduced incidence of side reactions found in aqueous media, and reduced microbial contamination [ 12 ]. Enzymes active in water miscible solvents are highly desired in biocatalysis where substrate solubility is limited and also when the solvent is desired as an acyl acceptor in transesterification reactions, as in the case of biodiesel production [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

Environmental Factors Modulating the Stability and Enzymatic Activity of the Petrotoga mobilis Esterase (PmEst)

et al. 2016

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Enzymes isolated from thermophilic organisms found in oil reservoirs can find applications in many fields, including the oleochemical, pharmaceutical, bioenergy, and food/dairy industries. In this study, in silico identification and recombinant production of an esterase from the extremophile bacteria Petrotoga mobilis (designated PmEst) were performed. Then biochemical, bioinformatics and structural characterizations were undertaken using a combination of synchrotron radiation circular dichroism (SRCD) and fluorescence spectroscopies to correlate PmEst stability and hydrolytic activity on different substrates. The enzyme presented a high Michaelis-Menten constant (KM 0.16 mM) and optimum activity at ~55°C for p-nitrophenyl butyrate. The secondary structure of PmEst was preserved at acid pH, but not under alkaline conditions. PmEst was unfolded at high concentrations of urea or guanidine through apparently different mechanisms. The esterase activity of PmEst was preserved in the presence of ethanol or propanol and its melting temperature increased ~8°C in the presence of these organic solvents. PmEst is a mesophilic esterase with substrate preference towards short-to medium-length acyl chains. The SRCD data of PmEst is in agreement with the prediction of an α/β protein, which leads us to assume that it displays a typical fold of esterases from this family. The increased enzyme stability in organic solvents may enable novel applications for its use in synthetic biology. Taken together, our results demonstrate features of the PmEst enzyme that indicate it may be suitable for applications in industrial processes, particularly, when the use of polar organic solvents is required.

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“…In recent years, esterases have been widely used in food production, detergent products, pharmaceuticals, perfumes, degradation of pollutants and the synthesis of optically pure compounds [ 4 , 5 ] owing to their broad array of substrate specificity and versatility in the reactions they catalyze [ 3 ]. Esterases with novel properties have a more and broad application prospect: cold-adapted esterases are applied to the industrial reaction at low temperature and benefit energy conservation [ 6 – 8 ]; salt-tolerant esterases are suitable for the reactions under a high salt concentration [ 9 – 11 ]; organic solvent-tolerant esterases/lipases are necessary for the substrates which are insoluble to water and the trans-esterification reaction in biodiesel production [ 5 , 12 , 13 ]. Nevertheless, there have been only a few reports available about these esterases with novel properties, implying the necessity to find more novel esterases.…”

Section: Introductionmentioning

confidence: 99%

A novel cold-adapted esterase from Enterobacter cloacae: Characterization and improvement of its activity and thermostability via the site of Tyr193Cys

Gao

Bandikari

et al. 2018

Microb Cell Fact

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BackgroundIn industries lipolytic reactions occur in insensitive conditions such as high temperature thus novel stout esterases with unique properties are attracts to the industrial application. Protein engineering is the tool to obtain desirable characters of enzymes. A novel esterase gene was isolated from South China Sea and subjected to a random mutagenesis and site directed mutagenesis for higher activity and thermo-stability compared to wild type.ResultsA novel esterase showed the highest hydrolytic activity against p-nitrophenyl acetate (pNPA, C2) and the optimal activity at 40 °C and pH 8.5. It was a cold-adapted enzyme and retained approximately 40% of its maximum activity at 0 °C. A mutant, with higher activity and thermo-stability was obtained by random mutagenesis. Kinetic analysis indicated that the mutant Val29Ala/Tyr193Cys shown 43.5% decrease in Km, 2.6-fold increase in Kcat, and 4.7-fold increase in Kcat/Km relative to the wild type. Single mutants V29A and Y193C were constructed and their kinetic parameters were measured. The results showed that the values of Km, Kcat, and Kcat/Km of V29A were similar to those of the wild type while Y193C showed 52.7% decrease in Km, 2.7-fold increase in Kcat, and 5.6-fold increase in Kcat/Km compared with the wild type. The 3-D structure and docking analysis revealed that the replacement of Tyr by Cys could enlarge the binding pocket. Moreover Y193C also showed a better thermo-stability for the reason its higher hydrophobicity and retained 67% relative activity after incubation for 3 h at 50 °C.ConclusionsThe superior quality of modified esterase suggested it has great potential application in extreme conditions and the mutational work recommended that important information for the study of esterase structure and function.Electronic supplementary materialThe online version of this article (10.1186/s12934-018-0885-z) contains supplementary material, which is available to authorized users.

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Esterase Active in Polar Organic Solvents from the YeastPseudozymasp. NII 08165

Cited by 13 publications

References 26 publications

Integrated strategy for purification of esterase from Aureobasidium pullulans

Integrated strategy for purification of esterase from Aureobasidium pullulans

Environmental Factors Modulating the Stability and Enzymatic Activity of the Petrotoga mobilis Esterase (PmEst)

A novel cold-adapted esterase from Enterobacter cloacae: Characterization and improvement of its activity and thermostability via the site of Tyr193Cys

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