Immobilization of Trypsin on Chitosan Nonwoven Using Glutaraldehyde

Kim, Jung Soo; Lee, So Hee; Song, Wha Soon

doi:10.5850/jksct.2013.37.7.852

Cited by 3 publications

(5 citation statements)

References 35 publications

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“…Chitosan nonwoven was pre-activated with glutaraldehyde at the optimal treatment conditions according to previous work [36]. A previous study indicated glutaraldehyde pretreatment conditions concerning pH, glutaraldehyde concentration, and crosslinking time for chitosan nonwovens to increase the activity of immobilized enzymes.…”

Section: Methodsmentioning

confidence: 99%

Immobilization of Trypsin from Porcine Pancreas onto Chitosan Nonwoven by Covalent Bonding

Kim

Lee

2019

Polymers

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The present study deals with the potential application of chitosan nonwoven for biomedical textiles based on enzyme immobilization. For this, chitosan nonwoven was first cross-linked with glutaraldehyde to introduce aldehyde groups at optimal conditions. To immobilize the enzyme trypsin onto glutaraldehyde-pre-activated chitosan nonwoven, several parameters such as pH, enzyme concentration, and reaction times were investigated. In addition, the pH, thermal stability, storage stability, and reusability of immobilized trypsin were examined. We found that the optimal immobilization conditions for trypsin were pH 8.5, enzyme concentration of 8% (owf), and treatment time of 30 min. Trypsin was immobilized at 25 °C efficiently. The immobilized trypsin showed lower pH stability and better thermal stability than free trypsin. The immobilized trypsin showed 50% of its initial activity after being used 15 times and 80% of that after 20 days of storage at 4 °C. SEM analysis also confirmed that trypsin was immobilized on chitosan nonwoven.

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Section: Methodsmentioning

confidence: 99%

Immobilization of Trypsin from Porcine Pancreas onto Chitosan Nonwoven by Covalent Bonding

Kim

Lee

2019

Polymers

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“…Optimization of the crosslinking conditions using GA on PET BSA Figure 5 shows the optimization of crosslinking conditions using GA with various pH values (a), GA concentrations (b) and crosslinking times (c). To optimize the pH of the crosslinking solution, the crosslinking time using GA was controlled to 120 min (Figure 5 (a)), according to Kim et al 23 At pH values lower than 8.0, the reaction between the aldehyde groups in GA and amino groups in enzymes decreased during the crosslinking process. 24 However, the relative activity increased at pH 8.0 or more, consistent with the results of a previous study 25 that reported that increasing the pH to obtain alkaline conditions promotes GA crosslinking and increases the amount of immobilized enzyme in a linear manner.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, the crosslinking time was shortened to 90 min in contrast to the 120 min used in a previous study. 23 Thus, the conditions for GA crosslinking on the PET BSA fabric were optimized to pH 10.0, 3% (v/v) GA, and 90 min crosslinking time. Hereafter, PET GA has been used to refer to samples crosslinked with GA under optimal conditions on PET BSA .…”

Section: Resultsmentioning

confidence: 99%

Development of an enzyme-immobilized support using a polyester woven fabric

Shim

Lee

Song

et al. 2016

Textile Research Journal

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In this study, we aimed to develop an enzyme-immobilized support using polyester woven fabrics and to optimize the development process. We obtained information about the storage stability and reusability of the enzyme and showed the applicability of the polyester woven fabric as an enzyme-immobilized support. In particular, the samples hydrolyzed by hydrogen chloride were treated with N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide to activate the surfaces. We evaluated the relative activity of the enzyme immobilization processes, the introduction of spacers, crosslinking and enzyme immobilization and optimized these parameters. The introduction process was controlled to a bovine serum albumin concentration of 1.5% (w/v) and treatment time of 3 h. The crosslinking process was optimized to pH 10.0, a glutaraldehyde concentration of 3% (v/v) and a crosslinking time of 90 min. The immobilization conditions were maintained at pH 8.5, a temperature of 25 C, a time of 45 min and a trypsin concentration of 6% (o.w.f.).Enzyme-immobilized supports have been used in various medical applications, food production processes, ion batteries and filters for waste-water purification. 1,2 Numerous methods for enzyme immobilization on different supports have been described in the literature. Ideal support properties include mechanical stability, hydrophilicity, resistance to microbial attack, porosity and availability at low cost. 2,3 The application of fabrics as immobilization supports could offer several advantages in terms of price and structure compared with other materials such as gels, beads, membranes and films. 2-4 Fabrics are commercially available and relatively cheaper than these or inorganic materials. The fiber-based structures have a high porosity and a large average pore size, so that more enzymes are immobilized, and the immobilized enzymes are protected from the environment. [2][3][4] There are various fabric supports available for enzyme immobilization, which can be classified into natural and synthetic fibers. Natural fibers such as cellulose and chitosan, which have excellent biocompatibility and hydrophilicity, are non-toxic and biodegradable. However, the weak mechanical stability of these natural fibers has greatly limited their application. 3,5 Synthetic fibers such as polylactic acid, polystyrene, polyamide and polyester (polyethylene terephthalate, PET) can be used for enzyme supports due to their good mechanical stability. However, their hydrophobicity is the major disadvantage. It is well known that the hydrophobic interaction between the enzyme and the support is one of the main reasons for protein denaturation. 3,5 One method to address these problems is to introduce spacers between the enzymes and the support. 6 This is a common method to reduce undesirable interactions between the functional groups of support surfaces and the enzyme molecule, and to lower steric hindrance. 7 In particular, the introduction of spacers increases the distance between the enzyme and the hydrophobic surfaces of sy...

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“…The amount of immobilized trypsin was determined by measuring absorbance at 410 nm with L-BAPNA as the substrate. 7,22,23 After the immobilization process, the recovered activity of the immobilized trypsin was calculated from the following equation: 7,24–28 where Ra is the activity recovery of the immobilized enzyme (%), A i is the activity of the immobilized enzyme (U), and A f is the activity of the same amount of free enzyme in solution as that immobilized on the support (U).…”

Section: Methodsmentioning

confidence: 99%

“…The activity of immobilized trypsin was measured each time using the method described above. 22,23,34…”

Section: Stability Assessmentmentioning

confidence: 99%

Covalent immobilization of enzyme on aminated woven poly (lactic acid) via ammonia plasma: evaluation of the optimum immobilization conditions

Song

Yeo

et al. 2016

Textile Research Journal

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The present study aims to develop an immobilization support from woven poly (lactic acid) (PLA) and establish the optimum immobilization conditions for trypsin. Woven PLA was modified by ammonia-based plasma treatment in order to incorporate amine groups on its surface. X-ray photoelectron spectroscopy analysis showed that the N1s composition of PLA increased significantly, from 0.66% to 5.92%, after ammonia-based plasma processing. Trypsin from porcine pancreas was immobilized onto modified woven PLA by covalent binding after activating PLA with glutaraldehyde (GA). The results indicated that the optimal GA treatment conditions were as follows: pH of 10.0, 2% GA (v/v), and 180 min crosslinking time. In addition, the optimum immobilization conditions were as follows: pH of 8.5, 10% (owf) of trypsin concentration, 30 min, and 25℃. Under the optimum conditions, the amount of immobilized enzyme on woven PLA was 0.28 mg/mg and specific activity was 3.763 U/mg. In addition, the pH and thermal stabilities of the immobilized trypsin were improved. The immobilized trypsin retained approximately 55% of its initial activity after 20 days of storage and exhibited the potential for repetitive use through approximately 15 cycles. GA crosslinking and trypsin immobilization were found to improve the roughness of the PLA surface and increase its hydrophobicity. The data indicate that modified woven PLA, used as an immobilization support, shows suitable properties for use as a biocatalytic material in enzymatic applications.

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Immobilization of Trypsin on Chitosan Nonwoven Using Glutaraldehyde

Cited by 3 publications

References 35 publications

Immobilization of Trypsin from Porcine Pancreas onto Chitosan Nonwoven by Covalent Bonding

Immobilization of Trypsin from Porcine Pancreas onto Chitosan Nonwoven by Covalent Bonding

Development of an enzyme-immobilized support using a polyester woven fabric

Covalent immobilization of enzyme on aminated woven poly (lactic acid) via ammonia plasma: evaluation of the optimum immobilization conditions

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