DNase-I-like enzyme from the carp liver—Inhibition by muscle and endogenous actin

Malicka-Błaszkiewicz, Maria; Majcher, Ilona; Nowak, Dorota

doi:10.1016/0020-711x(94)90137-6

Cited by 5 publications

(2 citation statements)

References 21 publications

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“…It is unclear why the CaMV 35S promoter fragment in the blood and muscle of carp consumed GM SBM diets could not be detected. The presence of DNase in the hepatopancreas of carp has been reported [20]. Krawczenko et al [21] reported that carp hepatopancreatic DNase has the ability to digest both native and denatured DNA.…”

Section: Discussionmentioning

confidence: 96%

Suitability of genetically modified soybean meal in a dietary ingredient for common carp Cyprinus carpio

et al. 2009

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The effect of genetically modified (GM) soybean meal (SBM) in a feed ingredient on growth performance of common carp was investigated in comparison to nonGM SBM. GM SBM was included at 34 and 48% in two experimental diets that were formulated with fish meal (FM) to obtain approximately 38% protein in diet. Two other experimental diets were formulated to contain the same levels of nonGM SBM. The diets were fed to juvenile common carp (22 g initial mean weight) for 12 weeks. The cauliflower mosaic virus (CaMV) 35S promoter fragment (205 base pairs) of the GM SBM was examined in fish muscle and blood samples at the twelfth week. From the twelfth week, the GM groups were fed with nonGM diets to determine the residual span of the transferred promoter fragment. There was no significant difference in growth and feed performance between GM and nonGM groups at two inclusion levels after 12 weeks. The CaMV 35S promoter fragment was not detected in fish muscles or blood receiving either level of GM SBM diet. The results demonstrated that the availability of GM SBM was similar to that of nonGM SBM and the GM SBM would be a suitable and safe ingredient in feed for common carp.

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Section: Discussionmentioning

confidence: 96%

Suitability of genetically modified soybean meal in a dietary ingredient for common carp Cyprinus carpio

et al. 2009

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“…Due to its high content of disulfide bonds and a highly cross-linked structure, shrimp DNase I is relatively stable to heat or SDS treatment [8]. A DNase-I-like enzyme, which is inhibited by muscle and liver actin, has been isolated from carp liver [9]. We investigated fish DWase 1 because fish are vertebrates but have a fused hepatopancreas similarly to shrimp.…”

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confidence: 99%

Purification and Characterization of Tilapia (Oreochromis mossambicus) Deoxyribonuclease I

Hsiao

Wang

et al. 1997

European Journal of Biochemistry

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DNase I of tilapia (Oreochromis rnossambicus) was purified to homogeneity. Tilapia DNase I is most active at pH 8.5 with Mg" as activator. The CaZ+/Mg2+ pair has a synergistic effect on activation. The enzyme is readily inactivated by heating above 55 "C, but is not inactivated by trypsin or 2-mercaptoethanol under alkaline conditions, with or without CaCI,. Its isoelectric point is 6.0. The 258-amino-acid sequence of tilapia DNase I was derived from overlapping sequences of tryptic, chymotryptic and CNBr peptides. The purified enzyme has two variants differing by a single Lys+Arg mutation at position 125. The polypeptide chain has one disulfide bridge and one carbohydrate side chain. By mass spectrometry, the purified enzyme shows many molecular mass forms differing by Lys/Arg substitution and sugar-chain length. [5]. The enzymatic properties of DNase I in these three species are very similar, and their primary structures are highly conserved [6]. It has been reported 171 that although the enzymatic properties of shrimp DNase I are very similar to those of mammalian DNase I, the protein structure is very different. Due to its high content of disulfide bonds and a highly cross-linked structure, shrimp DNase I is relatively stable to heat or SDS treatment [8]. A DNase-I-like enzyme, which is inhibited by muscle and liver actin, has been isolated from carp liver [9]. We investigated fish DWase 1 because fish are vertebrates but have a fused hepatopancreas similarly to shrimp. RNA was isolated from a piece of fresh small intestine of the fish with TRIzol Reagent (Life Technologies). Iodoacetic acid and P-mercaptoethanol were obtained from Wako. DEAE-cellulose (DE-52) was from Whatman. Hydroxyapatite (DNA grade) was from Bio-Rad. Trypsin and phenyl-Sepharose 4B were from Sigma. Sephadex G-I 00, ampholytes and concanavalin-A-Sepharose were from Pharmacia. Purified bovine DNase I was obtained via the method described previously [lo]. MATERIALS AND METHODS MaterialsDNase I assay. DNase I activity was determined by the hyperchromicity method [3, 101. 1 U enzyme activity causes an increase in absorbance of 1 at 260 nm in a 1-cm cuvette, per min per ml of assay mixture at 25°C.Purification of tilapia DNase 1. Approximately 450 g of tilapia hepatopancreas was minced and homogenized with a homogenizer (Heidolph DIAX 600) in 400ml 20mM TrislHCl, pH 8.0. Cell debris were removed by centrifugation in a Beckman JA14 rotor at 15 000 rpm for 1 h. The supernatant was filtered through two layers of cheese cloth, and the clear solution, termed crude extract, was subjected to (NH,),SO, fractionation between 35 % and 65 70 saturation. The precipitate obtained was dissolved in SO ml 20 mM Tris/HCl, pH 8.0, and dialyzed against the same buffer for 12 h. The dialyzed solution was passed through a DEAE-cellulose column (5 cmX15 cm), and the column was washed with 900ml of the same buffer. The absorbed proteins were eluted with a linear gradient of NaCl from 0 to 0.2 M at a flow rate of 60 ml/h. DNase I was eluted in approximately 0.05 M ...

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The Detection of Enzyme Activity Following Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis

Bischoff

Shi

Kennelly

1998

Analytical Biochemistry

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DNase-I-like enzyme from the carp liver—Inhibition by muscle and endogenous actin

Cited by 5 publications

References 21 publications

Suitability of genetically modified soybean meal in a dietary ingredient for common carp Cyprinus carpio

Suitability of genetically modified soybean meal in a dietary ingredient for common carp Cyprinus carpio

Purification and Characterization of Tilapia (Oreochromis mossambicus) Deoxyribonuclease I

The Detection of Enzyme Activity Following Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis

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