Immobilization of Thiol Proteases onto Porous Poly(vinyl alcohol) Beads

Hayashi, Toshio; Hyon, Suong‐Hyu; Cha, Wol-Suk; Ikada, Yoshito

doi:10.1295/polymj.25.489

Cited by 20 publications

(12 citation statements)

References 18 publications

(8 reference statements)

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“…Figure 5 shows that the pH range at which the immobilized enzyme had high activity (>50%) was considerably widened compared to that of the free one, probably due to diffusional limitations of the immobilized enzyme molecules. Similar behavior has been described for the immobilization of papain on poly(vinyl alcohol) beads activated by hexamethylene diisocyanate (28). Additionally, a very significant activity change for free enzyme, at pH 8, was observed depending on buffer composition ( Figure 5A).…”

Section: Immobilization Conditions Using Alginate-glutaraldehydesupporting

confidence: 78%

Neutrase Immobilization on Alginate−Glutaraldehyde Beads by Covalent Attachment

Ortega

Pérez-Mateos

Pilar

et al. 2008

J. Agric. Food Chem.

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Neutrase, a commercial preparation of Bacillus subtilis , was covalently immobilized on alginate-glutaraldehyde beads. Immobilization conditions and characterization of the immobilized enzyme were investigated. Central composite design and response surface methods were employed to evaluate the effects of immobilization parameters, such as glutaraldehyde concentration, enzyme loading, immobilization pH, and immobilization time. Under optimized working conditions (2% alginate, 6.2% glutaraldehyde, 61.84 U mL(-1) Neutrase, pH 6.2, and 60 min) the immobilization yield was about 50%. The immobilized enzyme exhibited higher K(m) compared to the soluble enzyme. The pH-activity profile was widened upon immobilization. The optimum temperature was shifted from 50 to 60 degrees C, and the apparent activation energy was decreased from 47.7 to 22.0 kJ mol(-1) by immobilization. The immobilized enzyme also showed significantly enhanced thermal stability.

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Section: Immobilization Conditions Using Alginate-glutaraldehydesupporting

confidence: 78%

Neutrase Immobilization on Alginate−Glutaraldehyde Beads by Covalent Attachment

Ortega

Pérez-Mateos

Pilar

et al. 2008

J. Agric. Food Chem.

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“…Proteases have been immobilized on various supports like gold colloids [8], carbohydrate polymers [9][10][11], polyvinyl alcohol beads [12] and microbial polysaccharides [13]. Recently attempts to immobilize protease on TiO 2 , magnetic, bimetallic Ag-Au, silica nanoparticles have also been successful [14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Immobilization of halophilic Bacillus sp. EMB9 protease on functionalized silica nanoparticles and application in whey protein hydrolysis

Sinha

Khare

2014

Bioprocess Biosyst Eng

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The present work targets the fabrication of an active, stable, reusable enzyme preparation using functionalized silica nanoparticles as an effective enzyme support for crude halophilic Bacillus sp. EMB9 protease. The immobilization efficiency under optimized conditions was 60%. Characterization of the immobilized preparation revealed marked increase in pH and thermal stability. It retained 80% of its original activity at 70 °C while t 1/2 at 50 °C showed a five-fold enhancement over that for the free protease. Kinetic constants K m and V max were indicative of a higher reaction velocity along with decreased affinity for substrate. The preparation could be efficiently reused up to 6 times and successfully hydrolysed whey proteins with high degree of hydrolysis. Immobilization of a crude halophilic protease on a nanobased scaffold makes the process cost effective and simple.

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“…1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 Immobilization of the enzyme papain using the two-step mechanism. Papain is a cysteineprotease from C. papaya and was selected for its potential applications in biotechnology [34][35][36] and biomedicine, i.e. as wound debriding formulations used for the treatment of diabetic ulcers or infected wounds 24,37 .…”

Section: Reaction Of 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide (Ementioning

confidence: 99%

TEMPO-Oxidized Nanofibrillated Cellulose as a High Density Carrier for Bioactive Molecules

et al. 2015

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Controlled and efficient immobilization of specific biomolecules is a key technology to introduce new, favorable functions to materials suitable for biomedical applications. Here, we describe an innovative and efficient, two-step methodology for the stable immobilization of various biomolecules, including small peptides and enzymes onto TEMPO oxidized nanofibrillated cellulose (TO-NFC). The introduction of carboxylate groups to NFC by TEMPO oxidation provided a high surface density of negative charges able to drive the adsorption of biomolecules and take part in covalent cross-linking reactions with 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDAC) and glutaraldehyde (Ga) chemistry. Up to 0.27 μmol of different biomolecules per mg of TO-NFC could be reversibly immobilized by electrostatic interaction. An additional chemical cross-linking step prevented desorption of more than 80% of these molecules. Using the cysteine-protease papain as model, a highly active papain-TO-NFC conjugate was achieved. Once papain was immobilized, 40% of the initial enzymatic activity was retained, with an increase in kcat from 213 to >700 s(-1) for the covalently immobilized enzymes. The methodology presented in this work expands the range of application for TO-NFC in the biomedical field by enabling well-defined hybrid biomaterials with a high density of functionalization.

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Immobilization of Thiol Proteases onto Porous Poly(vinyl alcohol) Beads

Cited by 20 publications

References 18 publications

Neutrase Immobilization on Alginate−Glutaraldehyde Beads by Covalent Attachment

Neutrase Immobilization on Alginate−Glutaraldehyde Beads by Covalent Attachment

Immobilization of halophilic Bacillus sp. EMB9 protease on functionalized silica nanoparticles and application in whey protein hydrolysis

TEMPO-Oxidized Nanofibrillated Cellulose as a High Density Carrier for Bioactive Molecules

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