Preparation and characterization of cross-linked enzyme aggregates (CLEAs) of recombinant poly-3-hydroxybutyrate depolymerase from Streptomyces exfoliatus

Hormigo, Daniel; García-Hidalgo, Javier; Acebal, Carmen; Mata, Isabel de la; Arroyo, Miguel

doi:10.1016/j.biortech.2011.09.035

Cited by 45 publications

(25 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The half-life of the CLEAs at 65 o C was 40 min, whereas that of the free enzyme was 23 min, which means that the thermostability of the CLEAs at 65 o C was improved by 1.7-fold compared to that of the free enzyme. Such thermostabilization of CLEAs-forms of other enzymes has been observed in many previous studies (22,23), and the stabilization of the enzyme conformation by rigidification of the enzyme structure through multipoint covalent attachment has been expected as the main cause for the stabilization of CLEAs (24,25).…”

Section: Resultsmentioning

confidence: 53%

See 1 more Smart Citation

Optimizing the preparation conditions and characterization of cross-linked enzyme aggregates of a monoamine oxidase

Kim

2016

Food Sci Biotechnol

View full text Add to dashboard Cite

Monoamine oxidases are useful in determination of biogenic monoamines, particularly histamine and tyramine. In this study, cross-linked enzyme aggregates (CLEAs) technique was applied to improve the stability of a monoamine oxidase from (AMAO). Under the optimized condition (50% of saturated ammonium sulfate, 5 mM glutaraldehyde, 2.0 mg/mL AMAO, 4 h-cross-linking at 25°C, pH 8.0), CLEAs-AMAO was recovered with a yield of 82% based on the subjected total enzyme activity. Both pH activity and stability at alkaline pHs of CLEAs-AMAO were significantly improved compared to those of the free enzyme, resulting in the shift of optimum pH to pH 8.0 and a broader pH profile. The half-life of the CLEAs at 65°C was elongated by 1.7-fold compared to that of the free enzyme, suggesting the thermal stability of AMAO was also improved by the CLEAs formation.

show abstract

Section: Resultsmentioning

confidence: 53%

“…3A). The shift of the optimal pHs and stabilization of the CLEAs at alkaline pHs are often reported in literatures (21)(22)(23).…”

Section: Resultsmentioning

confidence: 78%

Optimizing the preparation conditions and characterization of cross-linked enzyme aggregates of a monoamine oxidase

Kim

2016

Food Sci Biotechnol

View full text Add to dashboard Cite

show abstract

“…The improved thermal stability presented by CLEAs was reported earlier. For example, the CLEAs of poly-3-hydroxybutyrate depolymerase retained 50% activity after incubation at 70 • C for 40 min, while the free enzyme was completely deactivated after incubation under the same condition [29]; the CLEAs of laccases from two fungal strains retained more than 50% of their activity at 50 • C, whereas free laccases rapidly decreased in activity after 40 • C [30]; the CLEAs of xylanase from Geobacillus thermodenitrificans X1 maintained 53% of activity after 4 h at 70 • C in comparison to free xylanase that retained only 15% of activity [10]. This enhancement of thermal stability CLEAs may be partially attributed to the covalent cross-linking among enzyme aggregates.…”

Section: Effect Of the Temperature And Ph On Activity And Stabilitymentioning

confidence: 99%

Characterization of Cross-Linked Enzyme Aggregates of the Y509E Mutant of a Glycoside Hydrolase Family 52 β-xylosidase from G. stearothermophilus

et al. 2021

View full text Add to dashboard Cite

Cross-linked enzyme aggregates (CLEAs) of the Y509E mutant of glycoside hydrolase family 52 β-xylosidase from Geobacillus stearothermophilus with dual activity of β-xylosidase and xylanase (XynB2Y509E) were prepared. Ammonium sulfate was used as the precipitant agent, and glutaraldehyde as cross-linking agent. The optimum conditions were found to be 90% ammonium sulfate, 12.5 mM glutaraldehyde, 3 h of cross-linking reaction at 25 °C, and pH 8.5. Under these (most effective) conditions, XynB2Y509E-CLEAs retained 92.3% of their original β-xylosidase activity. Biochemical characterization of both crude and immobilized enzymes demonstrated that the maximum pH and temperature after immobilization remained unchanged (pH 6.5 and 65 °C). Moreover, an improvement in pH stability and thermostability was also found after immobilization. Analysis of kinetic parameters shows that the Km value of XynB2Y509E-CLEAs obtained was slightly higher than that of free XynB2Y509E (1.2 versus 0.9 mM). Interestingly, the xylanase activity developed by the mutation was also conserved after the immobilization process.

show abstract

“…Whereas, methodology of CLEA does not require purified enzymes, as it involve cross-linking of precipitated enzymes which need not be in it's pure form (Aytar and Bakir, 2008;Pan et al, 2011;Talekar et al, 2012a;Talekar et al, 2012b). The general methodology of CLEA has been widely used in various earlier studies (Agyei and He, 2015;Cui et al, 2014;García-García et al, 2012;Hormigo et al, 2012;Maria et al, 2011;Pan et al, 2011;Talekar et al, 2012a;Vaidya et al, 2012;Wang et al, 2011) and is also used in this study.…”

Section: Introductionmentioning

confidence: 99%

Solid state fermentation with recovery of Amyloglucosidase from extract by direct immobilization in cross linked enzyme aggregate for starch hydrolysis

Gupta

Jana

Kumar

et al. 2015

Biocatalysis and Agricultural Biotechnology

View full text Add to dashboard Cite

Preparation and characterization of cross-linked enzyme aggregates (CLEAs) of recombinant poly-3-hydroxybutyrate depolymerase from Streptomyces exfoliatus

Cited by 45 publications

References 30 publications

Optimizing the preparation conditions and characterization of cross-linked enzyme aggregates of a monoamine oxidase

Optimizing the preparation conditions and characterization of cross-linked enzyme aggregates of a monoamine oxidase

Characterization of Cross-Linked Enzyme Aggregates of the Y509E Mutant of a Glycoside Hydrolase Family 52 β-xylosidase from G. stearothermophilus

Solid state fermentation with recovery of Amyloglucosidase from extract by direct immobilization in cross linked enzyme aggregate for starch hydrolysis

Contact Info

Product

Resources

About