Spacer effects on enzymatic activity of bromelain immobilized onto porous chitosan beads

Seo, Hiroshi; Itoyama, Koki; Morimoto, Keiji; Takagishi, Toru; Oka, M.; Hayashi, Toshio

doi:10.1016/s0014-3057(97)00213-9

Cited by 18 publications

(4 citation statements)

References 14 publications

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“…Thus, we will only briefly mention these methods but refer the interested reader to the extensive body of literature available. Common methods to covalently conjugate proteins to chitosan include methods based on glutaraldehyde [152], carbodiimide [153,154], glyoxal [155] and epoxy [156] chemistries. Non-covalent methods or a combination of covalent and non-covalent methods have also been used to assemble proteins to chitosan films.…”

Section: Biofunctionalizing Electrodeposited Chitosan Filmsmentioning

confidence: 99%

Chitosan to Connect Biology to Electronics: Fabricating the Bio-Device Interface and Communicating Across This Interface

et al. 2014

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Individually, advances in microelectronics and biology transformed the way we live our lives. However, there remain few examples in which biology and electronics have been interfaced to create synergistic capabilities. We believe there are two major challenges to the integration of biological components into microelectronic systems: (i) assembly of the biological components at an electrode address, and (ii) communication between the assembled biological components and the underlying electrode. Chitosan OPEN ACCESSPolymers 2015, 7 2 possesses a unique combination of properties to meet these challenges and serve as an effective bio-device interface material. For assembly, chitosan's pH-responsive film-forming properties allow it to "recognize" electrode-imposed signals and respond by self-assembling as a stable hydrogel film through a cathodic electrodeposition mechanism. A separate anodic electrodeposition mechanism was recently reported and this also allows chitosan hydrogel films to be assembled at an electrode address. Protein-based biofunctionality can be conferred to electrodeposited films through a variety of physical, chemical and biological methods. For communication, we are investigating redox-active catechol-modified chitosan films as an interface to bridge redox-based communication between biology and an electrode. Despite significant progress over the last decade, many questions still remain which warrants even deeper study of chitosan's structure, properties, and functions.

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Section: Biofunctionalizing Electrodeposited Chitosan Filmsmentioning

confidence: 99%

Chitosan to Connect Biology to Electronics: Fabricating the Bio-Device Interface and Communicating Across This Interface

et al. 2014

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“…However, the described immobilization methods are in some cases rather elaborate and often result in only a monolayer of enzyme having a limited effective activity. It is also suggested that enzyme immobilization affects enzyme activity and stability (Almeida et al, 1993;Danilich et al, 1993;Camacho-Rubio et al, 1996;Seo et al, 1998). Moreover, immobilization techniques are not always completely understood and sometimes not reproducible with respect to resulting enzyme activity, enzyme leakage, etc.…”

Section: Introductionmentioning

confidence: 99%

A flow-through amperometric sensor based on dialysis tubing and free enzyme reactors

Böhm

Pijanowska

Olthuis

et al. 2001

Biosensors and Bioelectronics

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“…Bromelain was covalently immobilized onto the surface of porous chitosan beads without glutaraldehyde [ 30 ], with and without alkyl chain spacers of different lengths. The relative activity of immobilized bromelain was found to be high toward a small ester substrate, N -benzyl- L -arginine ethyl ester (BAEE), but rather low toward casein, a high-molecular-weight substrate [ 31 ].…”

Section: Introductionmentioning

confidence: 99%

Novel Biocatalysts Based on Bromelain Immobilized on Functionalized Chitosans and Research on Their Structural Features

et al. 2022

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Enzyme immobilization on various carriers represents an effective approach to improve their stability, reusability, and even change their catalytic properties. Here, we show the mechanism of interaction of cysteine protease bromelain with the water-soluble derivatives of chitosan—carboxymethylchitosan, N-(2-hydroxypropyl)-3-trimethylammonium chitosan, chitosan sulfate, and chitosan acetate—during immobilization and characterize the structural features and catalytic properties of obtained complexes. Chitosan sulfate and carboxymethylchitosan form the highest number of hydrogen bonds with bromelain in comparison with chitosan acetate and N-(2-hydroxypropyl)-3-trimethylammonium chitosan, leading to a higher yield of protein immobilization on chitosan sulfate and carboxymethylchitosan (up to 58 and 65%, respectively). In addition, all derivatives of chitosan studied in this work form hydrogen bonds with His158 located in the active site of bromelain (except N-(2-hydroxypropyl)-3-trimethylammonium chitosan), apparently explaining a significant decrease in the activity of biocatalysts. The N-(2-hydroxypropyl)-3-trimethylammonium chitosan displays only physical interactions with His158, thus possibly modulating the structure of the bromelain active site and leading to the hyperactivation of the enzyme, up to 208% of the total activity and 158% of the specific activity. The FTIR analysis revealed that interaction between N-(2-hydroxypropyl)-3-trimethylammonium chitosan and bromelain did not significantly change the enzyme structure. Perhaps this is due to the slowing down of aggregation and the autolysis processes during the complex formation of bromelain with a carrier, with a minimal modification of enzyme structure and its active site orientation.

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Spacer effects on enzymatic activity of bromelain immobilized onto porous chitosan beads

Cited by 18 publications

References 14 publications

Chitosan to Connect Biology to Electronics: Fabricating the Bio-Device Interface and Communicating Across This Interface

Chitosan to Connect Biology to Electronics: Fabricating the Bio-Device Interface and Communicating Across This Interface

A flow-through amperometric sensor based on dialysis tubing and free enzyme reactors

Novel Biocatalysts Based on Bromelain Immobilized on Functionalized Chitosans and Research on Their Structural Features

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