Immobilization of trypsin on chitosan gels: Use of different activation protocols and comparison with other supports

Manrich, Anny; Galvão, Célia M.A.; Jesus, Charles D.F.; Giordano, Roberto; Giordano, Raquel de Lima Camargo

doi:10.1016/j.ijbiomac.2007.11.007

Cited by 46 publications

(22 citation statements)

References 20 publications

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“…This behavior may be the cause of the lower RA observed for all cases where GLU was used, as shown in Table 1. Besides, it has been reported that glutaraldehyde can also react with other groups of the enzyme [19]. In consequence, higher amounts of protein could be immobilized on supports activated with Table 1), but the immobilized enzyme activity and the enzyme recovery were higher for derivatives obtained using glyoxyl groups (GLY and EPI).…”

Section: Resultsmentioning

confidence: 99%

“…It has natural reactive amine groups, is biocompatible, is available in various forms (hydrogel, membrane, fiber, and film), is nontoxic, and is readily susceptible to chemical modification [3,14]. The interest in modifying chitosan using different activation agents, and the use of polyelectrolyte complexes, have arisen recently mostly for applications in the field of enzyme immobilization [1,19,28]. Several biopolymers have been used to prepare chitosan hybrid hydrogels, such as gelatin, collagen, carrageenan, alginate, polyvinyl alcohol (PVA), and carboxymethylcellulose (CMC) [3].…”

Section: Introductionmentioning

confidence: 99%

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Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts

Mendes

Castro

Rodrigues

et al. 2010

J Ind Microbiol Biotechnol

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This work aimed at the production of stabilized derivatives of Thermomyces lanuginosus lipase (TLL) by multipoint covalent immobilization of the enzyme on chitosan-based matrices. The resulting biocatalysts were tested for synthesis of biodiesel by ethanolysis of palm oil. Different hydrogels were prepared: chitosan alone and in polyelectrolyte complexes (PEC) with κ-carrageenan, gelatin, alginate, and polyvinyl alcohol (PVA). The obtained supports were chemically modified with 2,4,6-trinitrobenzene sulfonic acid (TNBS) to increase support hydrophobicity, followed by activation with different agents such as glycidol (GLY), epichlorohydrin (EPI), and glutaraldehyde (GLU). The chitosan-alginate hydrogel, chemically modified with TNBS, provided derivatives with higher apparent hydrolytic activity (HA(app)) and thermal stability, being up to 45-fold more stable than soluble lipase. The maximum load of immobilized enzyme was 17.5 mg g(-1) of gel for GLU, 7.76 mg g(-1) of gel for GLY, and 7.65 mg g(-1) of gel for EPI derivatives, the latter presenting the maximum apparent hydrolytic activity (364.8 IU g(-1) of gel). The three derivatives catalyzed conversion of palm oil to biodiesel, but chitosan-alginate-TNBS activated via GLY and EPI led to higher recovered activities of the enzyme. Thus, this is a more attractive option for both hydrolysis and transesterification of vegetable oils using immobilized TLL, although industrial application of this biocatalyst still demands further improvements in its half-life to make the enzymatic process economically attractive.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts

Mendes

Castro

Rodrigues

et al. 2010

J Ind Microbiol Biotechnol

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“…To end the reaction, sodium borohydride was added to the reaction up to a final concentration of 1 mg/mL and incubated for further 30 min under stirring [11]. Finally, the immobilized preparation was washed with an excess of 25 mM sodium phosphate, pH 7, and stored at 4°C until further use.…”

Section: Immobilization Of Xylanasesmentioning

confidence: 99%

Immobilization and Stabilization of Xylanase by Multipoint Covalent Attachment on Agarose and on Chitosan Supports

Manrich

Komesu

Adriano

et al. 2010

Appl Biochem Biotechnol

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Xylanases have important applications in industry. Immobilization and stabilization of enzymes may allow their reuse in many cycles of the reaction, decreasing the process costs. This work proposes the use of a rational approach to obtain immobilized commercial xylanase biocatalysts with optimized features. Xylanase NS50014 from Novozymes was characterized and immobilized on glyoxyl-agarose, agarose-glutaraldehyde, and agarose-amino-epoxy support and on differently activated chitosan supports: glutaraldehyde-chitosan, glyoxyl-chitosan, and epoxy-chitosan. Two different chitosan matrices were tested. The best chitosan derivative was epoxy-chitosan-xylanase, which presented 100% of immobilization yield and 64% of recovered activity. No significant increase on the thermal stability was observed for all the chitosan-enzyme derivatives. Immobilization on glyoxyl-agarose showed low yield immobilization and stabilization degrees of the obtained derivative. The low concentration of lysine groups in the enzyme molecule could explain these poor results. The protein was then chemically modified with ethylenediamine and immobilized on glyoxyl-agarose. The new enzyme derivatives were 40-fold more stable than the soluble, aminated, and dialyzed enzyme (70 degrees C, pH 7), with 100% of immobilization yield. Therefore, the increase of the number of amine groups in the enzyme surface was confirmed to be a good strategy to improve the properties of immobilized xylanase.

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“…Then, Schiff's bases were reduced using a 120 mM NaBH 4 final concentration at 25 °C during 30 min under agitation and beads were rinsed with distilled water, followed by 50 mM phosphate buffer (pH 7.0) and 0.01 % (w/v) sodium azide. Finally, derivatives were vacuum dried and stored at 4 °C [36].…”

Section: Glyoxyl-agarose L-arabinose Isomerase Immobilizationmentioning

confidence: 99%

Chemical improvement of chitosan-modified beads for the immobilization of Enterococcus faecium DBFIQ E36 l-arabinose isomerase through multipoint covalent attachment approach

Manzo

Sousa

Fenoglio

et al. 2015

Journal of Industrial Microbiology and Biotechnology

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D-tagatose is produced from D-galactose by the enzyme L-arabinose isomerase (L-AI) in a commercially viable bioprocess. An active and stable biocatalyst was obtained by modifying chitosan gel structure through reaction with TNBS, D-fructose or DMF, among others. This led to a significant improvement in L-AI immobilization via multipoint covalent attachment approach. Synthetized derivatives were compared with commercial supports such as Eupergit(®) C250L and glyoxal-agarose. The best chitosan derivative for L-AI immobilization was achieved by reacting 4 % (w/v) D-fructose with 3 % (w/v) chitosan at 50 °C for 4 h. When compared to the free enzyme, the glutaraldehyde-activated chitosan biocatalyst showed an apparent activity of 88.4 U g (gel) (-1) with a 211-fold stabilization factor while the glyoxal-agarose biocatalyst gave an apparent activity of 161.8 U g (gel) (-1) with an 85-fold stabilization factor. Hence, chitosan derivatives were comparable to commercial resins, thus becoming a viable low-cost strategy to obtain high active L-AI insolubilized derivatives.

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Immobilization of trypsin on chitosan gels: Use of different activation protocols and comparison with other supports

Cited by 46 publications

References 20 publications

Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts

Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts

Immobilization and Stabilization of Xylanase by Multipoint Covalent Attachment on Agarose and on Chitosan Supports

Chemical improvement of chitosan-modified beads for the immobilization of Enterococcus faecium DBFIQ E36 l-arabinose isomerase through multipoint covalent attachment approach

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