Parameters in preparation and characterization of cross linked enzyme aggregates (CLEAs)

Talekar, Sachin; Joshi, Asavari; Joshi, Gandhali; Kamat, Priyanka; Haripurkar, Rutumbara; Kambale, Shashikant

doi:10.1039/c3ra40818c

Cited by 204 publications

(152 citation statements)

References 170 publications

(202 reference statements)

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“…Talekar et al (2012) stabilized alpha-amylase from Bacillus amyloliquefaciens immobilized using (NH4)2SO4. As the concentration of this salt increased the enzyme activity also increased.…”

Section: Formulation Of Concentrated Metabolic Liquid With Cutinolytimentioning

confidence: 99%

Production of heterologous cutinases by E. coli and improved enzyme formulation for application on plastic degradation

Gomes¹,

Matamá²,

Cavaco‐Paulo³

et al. 2013

Electron. J. Biotechnol.

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Background: The hydrolytic action of cutinases has been applied to the degradation of plastics. Polyethylene terephthalate (PET) have long half-life which constitutes a major problem for their treatment as urban solid residues. The aim of this work was to characterize and to improve stable the enzyme to optimize the process of degradation using enzymatic hydrolysis of PET by recombinant cutinases. Results:The wild type form of cutinase from Fusarium solani pisi and its C-terminal fusion to cellulose binding domain N1 from Cellulomonas fimi were produced by genetically modified Escherichia coli. The maximum activity of cutinases produced in Lactose Broth in the presence of ampicillin and isopropyl β-D-1-thiogalactopyranoside (IPTG) was 1.4 IU/mL. Both cutinases had an optimum pH around 7.0 and they were stable between 30 and 50ºC during 90 min. The addition of glycerol, PEG-200 and (NH4)2SO4 to the metabolic liquid, concentrated by ultra filtration, stabilized the activity during 60 days at 28ºC. The treatment of PET with cutinases during 48 hrs led to maxima weight loss of 0.90%. Conclusions: Recombinant microbial cutinases may present advantages in the treatment of poly(ethylene terephthalate) PET through enzymatic treatments.

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“…Talekar et al (2012) stabilized alpha-amylase from Bacillus amyloliquefaciens immobilized using (NH4)2SO4. As the concentration of this salt increased the enzyme activity also increased.…”

Section: Formulation Of Concentrated Metabolic Liquid With Cutinolytimentioning

confidence: 99%

Production of heterologous cutinases by E. coli and improved enzyme formulation for application on plastic degradation

Gomes¹,

Matamá²,

Cavaco‐Paulo³

et al. 2013

Electron. J. Biotechnol.

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“…In the case of CLEA-β-SPI-12, a small shift to lower temperature (approximately 5 °C) of the maximum activity temperature was observed. It was expected that optimal pH and temperature of enzymes immobilized as CLEAs were shifted because covalent bonds established between amino groups of the enzyme and glutaraldehyde, changing the microenvironment and the conformational flexibility of the enzyme [49]. Talekar et al [72] reported a shift to higher temperature (around 15 °C) for β-amylase from Bacillus amyloliquefaciens after immobilization by CLEA technique.…”

Section: Effects Of Ph and Temperature On The Activity And The Stabilmentioning

confidence: 99%

“…It was expected that optimal pH and temperature of enzymes immobilized as CLEAs were shifted because covalent bonds established between amino groups of the enzyme and glutaraldehyde, changing the microenvironment and the conformational flexibility of the enzyme [49]. Talekar et al [72] reported a shift to higher temperature (around 15 °C) for β-amylase from Bacillus amyloliquefaciens after immobilization by CLEA technique. However, Mafra et al [58] a small shift to lower temperature (approximately 5 • C) of the maximum activity temperature was observed.…”

Section: Effects Of Ph and Temperature On The Activity And The Stabilmentioning

confidence: 99%

“…As explained in introduction, this may have a double positive effect: to improve the crosslinking step and to reduce the diffusion problems by diluting the enzyme into inert protein [46,48,49]. Thus, the effects of the feeder proteins (BSA or SPI) and glutaraldehyde concentrations for the same β-amylase concentration on the global yield were studied (Section 3.4).…”

Section: Optimization Of β-Amylase Immobilization Using Cleas Technologymentioning

confidence: 99%

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Maltose Production Using Starch from Cassava Bagasse Catalyzed by Cross-Linked β-Amylase Aggregates

et al. 2018

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Barley β-amylase was immobilized using different techniques. The highest global yield was obtained using the cross-linked enzyme aggregates (CLEA) technique, employing bovine serum albumin (BSA) or soy protein isolate (SPI) as feeder proteins to reduce diffusion problems. The CLEAs produced using BSA or SPI showed 82.7 ± 5.8 and 53.3 ± 2.4% global yield, respectively, and a stabilization effect was observed upon immobilization at neutral pH value, e.g., after 12 h at 55 • C, the free β-amylase is fully inactivated, while CLEAs retained 25 and 15% of activity (using BSA and SPI, respectively). CLEA using SPI was selected because of its easier recovery, being chosen to convert the residual starch contained in cassava bagasse into maltose. This biocatalyst permitted to reach almost 70% of maltose conversion in 4 h using 30.0 g/L bagasse starch solution (Dextrose Equivalent of 15.88) and 1.2 U of biocatalyst per gram of starch at pH 7.0 and 40 • C. After 4 reuses (batches of 12 h) the CLEA using SPI maintained 25.50 ± 0.01% of conversion due to the difficulty of recovering.

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“…This approach provided a simple strategy for obtaining excellent enzyme immobilization rate and desired stability for various biotechnological applications 7,8 . Moreover, glutaraldehyde was also exploited for preparing crosslinked enzyme aggregates (CLEAs) mainly in case of enzymes that have low density of surface reactive groups and therefore may pose problem to obtain a final solid catalyst 9,10 . The process of CLEAs formation involves the treatment of enzymes immobilized on supports that are non-reactive to glutaraldehyde in order to prevent enzyme leakage followed by the crosslinking of enzymes adsorbed on aminated supports.…”

Section: Fig 1: Chemical Properties Of Glutaraldehydementioning

confidence: 99%

Role of Glutaraldehyde in Imparting Stability to Immobilized β-Galactosidase Systems

Satar

Jafri

Rasool

et al. 2018

Braz. arch. biol. technol.

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This review article highlights the role of glutaraldehyde as a matrix activator/stabilizer in imparting higher operational and thermal stability to β-galactosidase (βG) for biotechnological applications. Glutaraldehyde has been used extensively as a crosslinking agent as well as for functionalization of matrices to immobilize β-galactosidase. Immobilized β-galactosidase systems (IβGS) obtained as a result of glutaraldehyde treatment has been employed to hydrolyze whey and milk lactose in batch reactors, continuous packed-bed and fluidized bed reactors under various operational conditions. Moreover, these IβGS have also been utilized for the production of galactooligosaccharides in food, dairy and fermentation industries. It was observed that glutaraldehyde provided

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Parameters in preparation and characterization of cross linked enzyme aggregates (CLEAs)

Abstract: Parameters in preparation and characterization of cross linked enzyme aggregates (CLEAs)

Cited by 204 publications

References 170 publications

Production of heterologous cutinases by E. coli and improved enzyme formulation for application on plastic degradation

Production of heterologous cutinases by E. coli and improved enzyme formulation for application on plastic degradation

Maltose Production Using Starch from Cassava Bagasse Catalyzed by Cross-Linked β-Amylase Aggregates

Role of Glutaraldehyde in Imparting Stability to Immobilized β-Galactosidase Systems

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