Immobilization of a recombinant thermostable esterase (Pf2001) from Pyrococcus furiosus on microporous polypropylene: Isotherms, hyperactivation and purification

Almeida, Rodrigo Volcan; Branco, Roberta Vieira; Peixoto, Bruno; Lima, Cíntia da Silva; Alquéres, Sylvia Maria Campbell; Martins, Orlando B.; Antunes, Octávio Augusto Ceva; Freire, Denise Maria Guimarães

doi:10.1016/j.bej.2007.09.019

Cited by 32 publications

(22 citation statements)

References 30 publications

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“…This result is higher than previously reported, a lipase extract from Geobacillus thermoleovarans immobilized in polypropylene resulted in 8.7 mg/g of adsorbed protein per support [7], and a lipase from Candida rugosa immobilized in polypropylene resulted in 8.1 mg/g of adsorbed protein per support [17]. A recombinant thermostable esterase (Pf2001) from Pyrococcus furiosus immobilized in polypropylene Accurel MP-1000 resulted in 35.5 mg/g of adsorbed protein per support [6]. According to Almeida et al (2008), there is a large variation of results depending on the adsorption system, where enzyme characteristics, support superficial area, and particle and pore size seem to play an important role in the adsorption process.…”

Section: Resultscontrasting

confidence: 69%

“…A recombinant thermostable esterase (Pf2001) from Pyrococcus furiosus immobilized in polypropylene Accurel MP-1000 resulted in 35.5 mg/g of adsorbed protein per support [6]. According to Almeida et al (2008), there is a large variation of results depending on the adsorption system, where enzyme characteristics, support superficial area, and particle and pore size seem to play an important role in the adsorption process. Effect of temperature on hydrolysis activity of free and immobilized enzyme was evaluated at 30, 40, 50, and 60°C (Fig.…”

Section: Resultsmentioning

confidence: 97%

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Immobilization of a Recombinant Esterase from Lactobacillus plantarum on Polypropylene Accurel MP1000

Kolling

Suguino

Brod

et al. 2010

Appl Biochem Biotechnol

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A recombinant esterase from Lactobacillus plantarum was immobilized on hydrophobic support polypropylene Accurel MP1000 by adsorption. Adsorption efficiency was 83%, and the immobilized protein was 12.4 mg/g of support. Esterase activity was determined using p-nitrophenyl butyrate as substrate, and highest activities were observed at 50 °C for immobilized enzyme and 30 °C for free enzyme extract. Concerning thermal stability, after enzyme incubation at 80 °C for 30 min, immobilized and free enzyme retained 91% and 56% of initial activity, respectively. Immobilized enzyme presented lower V(max) and higher K(m) than free enzyme. Protein was not released from the support, and esterase activity increased after 3 cycles of reuse.

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Section: Resultscontrasting

confidence: 69%

Section: Resultsmentioning

confidence: 97%

Immobilization of a Recombinant Esterase from Lactobacillus plantarum on Polypropylene Accurel MP1000

Kolling

Suguino

Brod

et al. 2010

Appl Biochem Biotechnol

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“…This enzyme was immobilized on microporous polypropylene at low ionic strength, showing the hyperactivation phenomenon [33]. In this work the same enzyme was immobilized on more hydrophobic supports showing, once more, the hyperactivation phenomenon.…”

Section: Discussionmentioning

confidence: 83%

Immobilization and Characterization of a Recombinant Thermostable Lipase (Pf2001) from Pyrococcus furiosus on Supports with Different Degrees of Hydrophobicity

et al. 2010

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We studied the immobilization of a recombinant thermostable lipase (Pf2001Δ60) from the hyperthermophilic archaeon Pyrococcus furiosus on supports with different degrees of hydrophobicity: butyl Sepabeads and octadecyl Sepabeads. The enzyme was strongly adsorbed in both supports. When it was adsorbed on these supports, the enzyme showed 140 and 237% hyperactivation, respectively. The assessment of storage stability showed that the octadecyl Sepabeads immobilized enzyme showed 100% of residual activity after 30 days of storage. However, the greatest stability at 70°C was obtained in butyl Sepabeads immobilized enzyme, which retained 77% activity after 1 hour incubation. The maximum activity of the immobilized preparations was obtained with the pH between 6 and 7, at 70°C. Thus, this study achieved a new extremophilic biocatalyst with greater stability, for use in several biotechnological processes.

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“…As a result, immobilization of NStcI was successfully performed with an immobilization efficiency of 81.94% (based on esterase activity), and an amount of adsorbed enzyme of 4 mg of adsorbed protein/g of support (Tables 1 and 2). Lipases are selectively adsorbed onto Accurel MP1000, a feature that results in the purification of the lipase as well as in its immobilization, due to their hydrophobic domains [6–9, 34]. May be we have obtained this behavior after adsorption of NStcI esterase as we have a low protein yield (42.48%), while immobilization efficiency was high (81.94%) (Table 1).…”

Section: Resultsmentioning

confidence: 92%

“…Adsorption of a protein onto a solid surface is a widely used method for enzyme immobilization. Different hydrolases have been immobilized on porous polypropylene Accurel [6–9]. …”

Section: Introductionmentioning

confidence: 99%

Immobilization and Biochemical Properties of the Enantioselective Recombinant NStcI Esterase ofAspergillus nidulans

et al. 2013

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The recombinant NStcI A. nidulans esterase was adsorbed on Accurel MP1000, where protein yield and immobilization efficiency were 42.48% and 81.94%, respectively. Storage stability test at 4°C and RT showed 100% of residual activity after 40 days at both temperatures. The biocatalyst retains more than 70% of its initial activity after 3 cycles of repeated use. Biochemical properties of this new biocatalyst were obtained. Maximum activity was achieved at pH 11 and 30°C, while the best stability was observed with the pH between 9 and 11 at 40°C. NStcI thermostability was increased after immobilization, as it retained 47.5% of its initial activity after 1 h at 60°C, while the free enzyme under the same conditions displayed no activity. NStcI preserved 70% of its initial activity in 100% hexane after 72 h. Enzymatic kinetic resolution of (R,S)-1-phenylethanol was chosen as model reaction, using vinyl acetate as acyl donor. After optimization of reaction parameters, the highest possible conversion (42%) was reached at 37°C, a w of 0.07, and 120 h of bioconversion in hexane with an enantiomeric excess of 71.7%. NStcI has selectivity for (R)-enantiomer. The obtained E value (31.3) is in the range considered useful to resolve enantiomeric mixtures.

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Immobilization of a recombinant thermostable esterase (Pf2001) from Pyrococcus furiosus on microporous polypropylene: Isotherms, hyperactivation and purification

Cited by 32 publications

References 30 publications

Immobilization of a Recombinant Esterase from Lactobacillus plantarum on Polypropylene Accurel MP1000

Immobilization of a Recombinant Esterase from Lactobacillus plantarum on Polypropylene Accurel MP1000

Immobilization and Characterization of a Recombinant Thermostable Lipase (Pf2001) from Pyrococcus furiosus on Supports with Different Degrees of Hydrophobicity

Immobilization and Biochemical Properties of the Enantioselective Recombinant NStcI Esterase ofAspergillus nidulans

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