Preparation and characterization of cross-linked enzyme aggregates (CLEAs) of recombinant thermostable alkylsulfatase (SdsAP) from Pseudomonas sp. S9

Shengping, Li; Y, Su; Liu, Yadan; Sun, Lifang; Yu, Meng‐Fei; Wu, Yunkun

doi:10.1016/j.procbio.2016.09.013

Cited by 20 publications

(16 citation statements)

References 33 publications

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“…The combination of enzyme and glutaraldehyde ties all available amino groups on the surface of the enzyme together; hence, the basic groups on the enzyme surface gave a positive charge to the enzyme protein and consequently shifted the optimum pH to the lower side. That is, an increase in the number of basic groups occurred after immobilization and the enzyme acquired a more polycationic character …”

Section: Resultsmentioning

confidence: 99%

“…That is, an increase in the number of basic groups occurred after immobilization and the enzyme acquired a more polycationic character. 29,30 For the determination of pH stability, the free enzyme and CLEAs were incubated in 100 mmol L −1 citrate and phosphate buffer at various pH values. As can be seen from Fig.…”

Section: Effect Of Ph On -Galactosidase Activity and Stabilitymentioning

confidence: 99%

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Cross‐linked α‐galactosidase aggregates: optimization, characterization and application in the hydrolysis of raffinose‐type oligosaccharides in soymilk

Bayraktar

Önal

2019

J Sci Food Agric

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BACKGROUND Cross‐linked enzyme aggregates (CLEAs) of α‐galactosidase, partially purified from maize (Zea mays) flour, were prepared. The impact of various parameters on enzyme activity was examined to optimize the immobilization procedure. Biochemical characterization of the free and immobilized enzyme was carried out. Stability (thermal, pH, storage and operational stability) and reusability tests were performed. The potential use of the free enzyme and the CLEAs in hydrolysis processes of raffinose‐type oligosaccharides present in soymilk was investigated. RESULTS α‐galactosidase CLEAs were prepared with 47% activity recovery under optimum conditions [1:5 (v/v) enzyme solution:saturated ammonium sulfate solution ratio; 7.5 mg protein and 0.1% (v/v) glutaraldehyde, 6 h, 4 °C, 150 rpm]. α‐galactosidase CLEAs exhibited increased stability in comparison to the free enzyme. The CLEAs and the free enzyme showed a maximum activity at 40°C and their optimal pH values were5.5 and 6.0, respectively. Kinetic constants (KM, Vmax and kcat) were calculated for the free enzyme and the CLEAs in the presence of p‐nitrophenyl‐α‐d‐galactopyranoside, stachyose, melibiose and raffinose. The effect of various chemicals and sugars on enzyme activity showed that both enzyme forms were significantly inhibited by HgCl2 and galactose. The CLEAs hydrolyzed 85% of raffinose and 96% of stachyose. CONCLUSION The α‐galactosidase CLEAs, with their satisfactory enzymatic characteristics, have much potential for use in the food and feed industry. © 2019 Society of Chemical Industry

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Section: Resultsmentioning

confidence: 99%

Section: Effect Of Ph On -Galactosidase Activity and Stabilitymentioning

confidence: 99%

Cross‐linked α‐galactosidase aggregates: optimization, characterization and application in the hydrolysis of raffinose‐type oligosaccharides in soymilk

Bayraktar

Önal

2019

J Sci Food Agric

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“…This bifunctional reagent responds with reactive NH 2 groups (mainly Lys) on the protein surface [ 11 ]. Glutaraldehyde is widely used as the cross-linking agent [ 28 ]. Varying concentrations of glutaraldehyde (2, 10, 20, 50, 100 mM) were used ( Table 1 ) [ 10 , 29 , 30 ].…”

Section: Methodsmentioning

confidence: 99%

Preparation and Optimisation of Cross-Linked Enzyme Aggregates Using Native Isolate White Rot Fungi Trametes versicolor and Fomes fomentarius for the Decolourisation of Synthetic Dyes

Vršanská

Voběrková

Jiménez

et al. 2017

IJERPH

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The key to obtaining an optimum performance of an enzyme is often a question of devising a suitable enzyme and optimisation of conditions for its immobilization. In this study, laccases from the native isolates of white rot fungi Fomes fomentarius and/or Trametes versicolor, obtained from Czech forests, were used. From these, cross-linked enzyme aggregates (CLEA) were prepared and characterised when the experimental conditions were optimized. Based on the optimization steps, saturated ammonium sulphate solution (75 wt.%) was used as the precipitating agent, and different concentrations of glutaraldehyde as a cross-linking agent were investigated. CLEA aggregates formed under the optimal conditions showed higher catalytic efficiency and stabilities (thermal, pH, and storage, against denaturation) as well as high reusability compared to free laccase for both fungal strains. The best concentration of glutaraldehyde seemed to be 50 mM and higher efficiency of cross-linking was observed at a low temperature 4 °C. An insignificant increase in optimum pH for CLEA laccases with respect to free laccases for both fungi was observed. The results show that the optimum temperature for both free laccase and CLEA laccase was 35 °C for T. versicolor and 30 °C for F. fomentarius. The CLEAs retained 80% of their initial activity for Trametes and 74% for Fomes after 70 days of cultivation. Prepared cross-linked enzyme aggregates were also investigated for their decolourisation activity on malachite green, bromothymol blue, and methyl red dyes. Immobilised CLEA laccase from Trametes versicolor showed 95% decolourisation potential and CLEA from Fomes fomentarius demonstrated 90% decolourisation efficiency within 10 h for all dyes used. These results suggest that these CLEAs have promising potential in dye decolourisation.

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“…Temperature was an important factor affecting enzyme activity. In Figure 5a, the free lipase showed high activity at a temperature range from 50 to 60 C and an optimal reaction temperature of 50 C. However, the CLEAs displayed high activity at a broader temperature range of alkylsulfatase-CLEAs, 29,30 while the optimal reaction temperature at lower temperature was different from the protease-CLEAs and alkylsulfatase-CLEAs. 29,31 Then we analyzed the thermal stability and measured the half-life (t 1/2 ) of the CLEAs.…”

Section: Optimum Temperature and Thermal Stabilitymentioning

confidence: 97%

Preparation and characterization of a nontoxic cross‐linked lipase aggregate by using sodium tripolyphosphate and chitosan

Yan

Shi

Ding

et al. 2021

J of Applied Polymer Sci

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Cross-linked enzyme aggregates (CLEAs) have the advantages of high-enzyme activity and stability, and have an application potential in food and feed processing. In order to address the food and feed safety of CLEAs and have ideal enzymatic properties, a new design of CLEAs of lipase from Aspergillus niger is developed by using sodium tripolyphosphate (TPP) as a cross-linking agent and chitosan as an auxiliary agent. The enzyme activity recovery of the developed CLEAs reaches 126%. The half-life of the CLEAs is 11.4-fold higher than that of free enzyme at the optimum reaction temperature of CLEAs. The surface structure of the CLEAs reveals cluster and porous structure. The variations of the secondary structures of the CLEAs indicate an increase of α-helix and decrease of β-sheet, β-turn, and random coil. The adding of chitosan is beneficial to enzyme activity recovery and thermal stability of the CLEAs. TPP and chitosan are safe and nontoxic, and the CLEAs prepared with TPP and chitosan is suitable for the food and feed industries.

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Preparation and characterization of cross-linked enzyme aggregates (CLEAs) of recombinant thermostable alkylsulfatase (SdsAP) from Pseudomonas sp. S9

Cited by 20 publications

References 33 publications

Cross‐linked α‐galactosidase aggregates: optimization, characterization and application in the hydrolysis of raffinose‐type oligosaccharides in soymilk

Cross‐linked α‐galactosidase aggregates: optimization, characterization and application in the hydrolysis of raffinose‐type oligosaccharides in soymilk

Preparation and Optimisation of Cross-Linked Enzyme Aggregates Using Native Isolate White Rot Fungi Trametes versicolor and Fomes fomentarius for the Decolourisation of Synthetic Dyes

Preparation and characterization of a nontoxic cross‐linked lipase aggregate by using sodium tripolyphosphate and chitosan

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