Identification of Antihyperuricemic Peptides in the Proteolytic Digest of Shark Cartilage Water Extract Using <i>in Vivo</i> Activity-Guided Fractionation

Murota, Itsuki; Taguchi, Satoko; Sato, Nobuyuki; Park, Eun Young; Nakamura, Yasushi; Satô, Kenji

doi:10.1021/jf405504u

Cited by 57 publications

(62 citation statements)

References 16 publications

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“…The type of bioactive peptides generated from a particular protein is dependent on the primary sequence of the protein and the specificity of proteases used to generate peptides [ 3 , 20 ]. Proteins and peptides from cartilages of Chondrichthyes have been investigated as a good source for searching antiangiogenesis, antioxidant, and antihyperuricemic agents [ 21 , 22 , 23 ]. Therefore, in the experiment, crude proteins of spotless smoothhound cartilage were extracted using HCl-Guanidine buffer with yield of 1.69% ± 0.03% (on dry cartilage), and hydrolyzed for 4 h using trypsin at pH 8.0, 40 °C with a total enzyme dose of 2.5%.…”

Section: Resultsmentioning

confidence: 99%

Bioactive Peptides from Cartilage Protein Hydrolysate of Spotless Smoothhound and Their Antioxidant Activity In Vitro

et al. 2018

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In the experiment, crude proteins from spotless smoothhound (Mustelus griseus), cartilages were isolated by HCl-Guanidine buffer, and its hydrolysate was prepared using trypsin at pH 8.0, 40 °C with a total enzyme dose of 2.5%. Subsequently, three antioxidant peptides were purified from the hydrolysate using membrane ultrafiltration, anion-exchange chromatography, gel filtration chromatography, and reverse phase high-performance liquid chromatography. The amino acid sequences of isolated peptides were identified as Gly-Ala-Glu-Arg-Pro (MCPE-A); Gly-Glu-Arg-Glu-Ala-Asn-Val-Met (MCPE-B); and Ala-Glu-Val-Gly (MCPE-C) with molecular weights of 528.57, 905.00, and 374.40 Da, respectively, using protein amino acid sequence analyzer and mass spectrum. MCPE-A, MCPE-B and MCPE-C exhibited good scavenging activities on 2,2-diphenyl-1-picrylhydrazyl radicals (DPPH•) (EC50 3.73, 1.87, and 2.30 mg/mL, respectively), hydroxyl radicals (HO•) (EC50 0.25, 0.34, and 0.06 mg/mL, respectively), 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid radicals (ABTS+•) (EC50 0.10, 0.05, and 0.07 mg/mL, respectively) and superoxide anion radicals (normalO2−•) (EC50 0.09, 0.33, and 0.18 mg/mL, respectively). MCPE-B showed similar inhibiting ability on lipid peroxidation with butylated hydroxytoluene (BHT) in a linoleic acid model system. Furthermore, MCPE-A, MCPE-B, and MCPE-C could protect H2O2-induced HepG2 cells from oxidative stress by decreasing the content of malonaldehyde (MDA) and increasing the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione reductase (GSH-Rx). Glu, Gly, Met, and Pro in their sequences and low molecular weight could be attributed to the antioxidant activities of three isolated peptides. These results suggested that GAERP (MCPE-A), GEREANVM (MCPE-B), and AEVG (MCPE-C) from cartilage protein hydrolysate of spotless smoothhound might serve as potential antioxidants and be used in the pharmaceutical and health food industries.

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Section: Resultsmentioning

confidence: 99%

Bioactive Peptides from Cartilage Protein Hydrolysate of Spotless Smoothhound and Their Antioxidant Activity In Vitro

et al. 2018

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“…The XO inhibition assay was carried out following the method of Murota et al . () with slight modifications. Fifty microlitre xanthine (6 mmol L −1 ), 50 μL XO (15 mU mL −1 ), 200 μL TPH, TPH‐M or TPH‐D were mixed in the wells of a 96‐well plate, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Murota et al . () found that Ser‐Pro‐Pro‐Tyr‐Trp‐Pro‐Tyr exhibited significantly different antihyperuricemic activity after oral administration or intravenous injection. In addition, during food processing and storage, Maillard reaction occurs spontaneously mainly through the interaction between reducing sugars and amino groups.…”

Section: Introductionmentioning

confidence: 98%

“…Interestingly, some individual amino acids (Val, Trp) and tryptophan‐containing peptides have been reported to exhibit negative effects on XO activity (Nongonierma & Fitzgerald, ; Nongonierma et al ., ). Besides, shark cartilage hydrolysate (Murota et al ., ) and walnut protein hydrolysate (Li et al ., ) showed antihyperuricemic activity after oral administration. Additionally, our previous study found that tuna protein hydrolysate (TPH) could effectively inhibit XO activity in vitro (Su et al ., ).…”

Section: Introductionmentioning

confidence: 99%

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Antihyperuricemic effect of tuna protein hydrolysate and derived products after in vitro digestion or Maillard reaction on oteracil potassium‐induced hyperuricemia rats

Yang

Zou

Zhao

et al. 2018

Int J of Food Sci Tech

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To investigate the effect of Maillard reaction (MR) and gastrointestinal digestion (GID) on the antihyperuricemic activity of tuna protein hydrolysate (TPH), hyperuricemia rats were treated with TPH, its MR products (TPH-M) and gastrointestinal digesta (TPH-D) in this work. Although the XO inhibitory activity of TPH-M was higher than TPH, TPH-M exhibited even lower serum-uric-acid-lowering activity than TPH. Additionally, there was no significant (P < 0.05) difference observed in the inhibition of XO activity in vitro and antihyperuricemic activity of TPH and TPH-D, indicated that the antihyperuricemic activity of TPH would not decrease after GID. Additionally, TPH, TPH-M and TPH-D could effectively inhibit XO activity and downregulate XO mRNA expression in vivo, suggesting the mechanisms of antihyperuricemic effect was related to XO inhibitory activity and XO mRNA expression. As a whole, TPH could be a useful functional ingredient against hyperuricemia.Antihyperuricemic effect of tuna hydrolysate and derived products Y. Liu et al. 264

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“…Obviously, non-specific proteases will release far more peptides. The traditional strategies involve assay-guided consecutive separation, including membrane filtration, isoelectric focusing, gel filtration, ion exchange and reverse phase chromatography, which need the analysis of activities fraction by fraction [ 18 , 19 , 20 , 21 ]_ENREF_14. Although these procedures are feasible, they are time-consuming and labor-intensive and give rise to problems such as poor isolation yields and loss of biological activity during operation.…”

Section: Introductionmentioning

confidence: 99%

Selective Extraction and Antioxidant Properties of Thiol-Containing Peptides in Soy Glycinine Hydrolysates

et al. 2018

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A highly selective procedure to extract thiol-containing peptides (TCPs) from complicated soy glycinin hydrolysates (SGHs) was described. This procedure included the reduction of disulfide bonds by 1,4-dithiothreitol (DTT) and enrichment of TCPs through Thiopropyl-Sephrose 6B covalent chromatography. TCPs were confirmed using a strategy based on mass shift after differential alkylation of sulfhydryl groups with iodoacetamide and N-ethylmaleimide by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF-MS). The antioxidant activities of TCPs were evaluated using chemical assays. DTT reduction increased the concentration of sulfhydryl groups from 1.8 μmol/g to 113.8 μmol/g. The efficiency of the extraction was improved by optimizing the loading of sample, extraction and desorption time and the content of desorption reagent. Both of the adsorption and desorption process were found to fit a pseudo-second order model. MALDI-TOF-MS showed that 36 of the 45 extracted peptides were TCPs. The EC50 of TCPs for DPPH, hydroxyl radical, and superoxide anion radical was 0.1, 1.49 and 0.084 mg/mL, respectively. The reducing power of TCPs (0.2 mg/mL) was of 0.375. These results suggest that the combination of DTT reduction and Thiopropyl-Sepharose 6B covalent chromatograph was a successful pathway to extract TCPs from SGHs and the TCPs could be used as potential antioxidants.

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Identification of Antihyperuricemic Peptides in the Proteolytic Digest of Shark Cartilage Water Extract Using in Vivo Activity-Guided Fractionation

Cited by 57 publications

References 16 publications

Bioactive Peptides from Cartilage Protein Hydrolysate of Spotless Smoothhound and Their Antioxidant Activity In Vitro

Bioactive Peptides from Cartilage Protein Hydrolysate of Spotless Smoothhound and Their Antioxidant Activity In Vitro

Antihyperuricemic effect of tuna protein hydrolysate and derived products after in vitro digestion or Maillard reaction on oteracil potassium‐induced hyperuricemia rats

Selective Extraction and Antioxidant Properties of Thiol-Containing Peptides in Soy Glycinine Hydrolysates

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