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Šafařı́k, Ivo; Ptáčková, L.; Koneracká, M.; Šafařı́ková, Mirka; Τimko, M.; Kopčanský, P.

doi:10.1023/a:1014521021795

Cited by 14 publications

(5 citation statements)

References 8 publications

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“…Heavy metals constitute a major group of aquatic pollutants and their influence on trypsin activity can be used as a tool for detecting xenobiotics [2,33]. Similar to other tropical fish proteases [1-4,6,10], crevalle jack trypsin showed sensitivity to metal ions, particularly Cd 2+ , Al 3+ , Zn 2+ , Cu 2+ , Pb 2+ , and Hg 2+ at 1 mM (Table 3).…”

Section: Resultsmentioning

confidence: 99%

Metal-sensitive and thermostable trypsin from the crevalle jack (Caranx hippos) pyloric caeca: purification and characterization

Costa

Júnior

Amaral

et al. 2013

Chemistry Central Journal

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BackgroundOver the past decades, the economic development and world population growth has led to increased for food demand. Increasing the fish production is considered one of the alternatives to meet the increased food demand, but the processing of fish leads to by-products such as skin, bones and viscera, a source of environmental contamination. Fish viscera have been reported as an important source of digestive proteases with interesting characteristics for biotechnological processes. Thus, the aim of this study was to purify and to characterize a trypsin from the processing by-products of crevalle jack (Caranx hippos) fish.ResultsA 27.5 kDa trypsin with N-terminal amino acid sequence IVGGFECTPHVFAYQ was easily purified from the pyloric caeca of the crevalle jack. Its physicochemical and kinetic properties were evaluated using N-α-benzoyl-DL-arginine-p-nitroanilide (BApNA) as substrate. In addition, the effects of various metal ions and specific protease inhibitors on trypsin activity were determined. Optimum pH and temperature were 8.0 and 50°C, respectively. After incubation at 50°C for 30 min the enzyme lost only 20% of its activity. K m , k cat, and k cat /K m values using BApNA as substrate were 0.689 mM, 6.9 s-1, and 10 s-1 mM-1, respectively. High inhibition of trypsin activity was observed after incubation with Cd2+, Al3+, Zn2+, Cu2+, Pb2+, and Hg2+ at 1 mM, revealing high sensitivity of the enzyme to metal ions.ConclusionsExtraction of a thermostable trypsin from by-products of the fishery industry confirms the potential of these materials as an alternative source of these biomolecules. Furthermore, the results suggest that this trypsin-like enzyme presents interesting biotechnological properties for industrial applications.

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Section: Resultsmentioning

confidence: 99%

Metal-sensitive and thermostable trypsin from the crevalle jack (Caranx hippos) pyloric caeca: purification and characterization

Costa

Júnior

Amaral

et al. 2013

Chemistry Central Journal

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“…Due to its narrow specificity in peptide cleavage almost exclusively directed toward L ‐lysyl and L ‐argininyl bonds,17 trypsin has been widely applied in various applications including the production of anti‐thrombotic peptides from milk casein and the manufacture of hypoallergenic infant food,18 and as a tool to identify and quantify xenobiotics 19. It has also been used in the identification of proteinaceous markers that define the progression and prognosis of human diseases,20 and frequently as an essential protease for peptide mapping and identification in proteomics.…”

Section: Introductionmentioning

confidence: 99%

Functionalization Strategies for Protease Immobilization on Magnetic Nanoparticles

Teoh

Gooding

et al. 2010

Adv Funct Materials

139

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A comprehensive study on the general functionalization strategies for magnetic nanoparticles (MNPs) is presented in this work. Using well‐established techniques as well as modified protocols, the wide range of functional moieties grafted on γ‐Fe2O3 (maghemite) nanosurfaces include those of amine, aldehyde, carboxylic, epoxy, mercapto, and maleimide ends. Among the modified protocols are the one‐step water‐catalyzed silanization with mercaptopropyltrimethoxysilane, resulting in dense distal thiols, and the direct functionalization with a heterogeneous bifunctional linker N‐[p‐maleimidophenyl]isocynanate (PMPI). The former results in a protective Stöber type coating while simultaneously reducing the iron oxide core to magnetite (Fe3O4). The conjugation of trypsin, hereby chosen as model biomolecule, onto the differently functionalized MNPs is further demonstrated and assessed based on its activity, kinetics, and thermo‐/long‐term stability as well as reusability. Besides aqueous stability and ease in recovery by magnetic separation, the immobilized trypsin on MNPs offers superior protease durability and reusability, without compromising the substrate specificity and sequence coverage of free trypsin. The MNP‐based proteases can be used as valuable carriers in proteomics and miniaturized total analysis devices. The applicability of the functional surfaces devised in the current study is also relevant for the conjugation of other biomolecules beyond trypsin.

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“…The possible clinical use of magnetically guided particles for drug delivery to tumours and elsewhere within the body has been studied for many years. There is a vast literature on the topic [1][2][3] and ongoing commercial development of various methods and materials for use in such therapies, e.g. [4][5][6], is taking place.…”

Section: Introductionmentioning

confidence: 99%

Magnetic guidance of ferrofluidic nanoparticles in anin vitromodel of intraocular retinal repair

2003

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Agarose gel at a concentration of 0.6% was used to simulate the vitreous body of the eye during the infusion of a ferrofluid and the subsequent magnetic concentration of it onto a surrogate retinal surface. The 10 nm Fe 3 O 4 particles in the ferrofluid served to mimic the cobalt particles in a silicone magnetic fluid that is being developed for use as a tamponading agent in magnetic fluid therapies designed to alleviate retinal detachments and other types of retinopathy. Magnetically guided interstitial diffusion of the nanoparticles through up to 20 mm of the gel over periods of 72 h was shown to be possible, thus demonstrating that essentially all points on the retinal surface are reachable from elsewhere in the ocular interior. The nanodynamics of the magnetic and viscous forces at work on the particles during movement through the gel are discussed; in particular the diffusion speeds of the particles are estimated and compared with observations.

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Cited by 14 publications

References 8 publications

Metal-sensitive and thermostable trypsin from the crevalle jack (Caranx hippos) pyloric caeca: purification and characterization

Metal-sensitive and thermostable trypsin from the crevalle jack (Caranx hippos) pyloric caeca: purification and characterization

Functionalization Strategies for Protease Immobilization on Magnetic Nanoparticles

Magnetic guidance of ferrofluidic nanoparticles in anin vitromodel of intraocular retinal repair

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