• Bulletin of the Korean Chemical Society
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• v.23 no.5
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• pp.647-654
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• 2002

The synthesis and characterization of third- and fifth-generation poly(propylene imine) dendrimers terminated with imidazole moieties is reported. Functionalization was achieved using simple peptide coupling reagents. These materials were characte rized by MALDI-MS, NMR, and titration. The use of these endgroup-functionalized dendrimers as catalysts for the hydrolysis of 2,4-dinitrophenyl acetate is described. Molecular simulations provide a basis for interpreting the catalytic data.

• Mass Spectrometry Letters
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• v.9 no.2
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• pp.46-50
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• 2018

The effects of temperature and acetonitrile (ACN) concentration on microwave-assisted weak-acid hydrolysis of proteins were investigated. Myoglobin was hydrolyzed for 1 h using 2% formic acid and a microwave with different concentrations of ACN (0, 5, and 10%) at various temperatures (50, 60, 70, 80, 90, and $100^{\circ}C$). The numbers of peptides identified with each concentration of ACN were the same for each temperature. The greatest number of peptides (18 total) was obtained with hydrolysis at $100^{\circ}C$, and 6 of these were a result of additional removal of aspartic acid at the C-terminus. Hydrolysis at $80^{\circ}C$ resulted in 13 peptides, of which only 1 was generated by the additional removal of aspartic acid, and 12 were observed with hydrolysis at $100^{\circ}C$. Our results demonstrate that microwave-assisted weak-acid hydrolysis of proteins can be performed successfully at $80^{\circ}C$, which could be beneficial for limiting side reactions and generating larger peptide sequences.

• Food Science of Animal Resources
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• v.29 no.5
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• pp.633-639
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• 2009

The principal objective of the current study was to prepare low molecular weight peptides from milk proteins using enzymatic hydrolysis techniques, in an effort to assess the antioxidant activity of these peptides. The casein and whey proteins isolated from fresh milk were treated with several proteolytic enzymes, such as chymotrypsin, pepsin, and trypsin and the resulting low molecular weight peptides were collected by TCA precipitation. Their identity was confirmed by SDS-PAGE analysis. The hydrolysis experiments indicated that whey protein treated with chymotrypsin displayed the highest degree of protein hydrolysis. The antioxidant activity of milk protein hydrolysates was determined by measuring the ABTS-radical scavenging activity. The results of these experiments showed that hydrolysis of the milk protein was effective in increasing their antioxidant activities. Especially, the tryptic digested casein displayed the highest radical scavenging activity (80.7%). The hydrolyzed low molecular weight milk protein was isolated using an ultrafiltration membrane. The casein hydrolysate passed through a membrane with molecular weight cut-off (MWCO) of 3 kDa displayed the strongest antioxidant activity.

• Applied Biological Chemistry
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• v.46 no.2
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• pp.130-133
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• 2003

A method for separating heme-iron from hemoglobin (Hgb) hydrolysate by dialysis was developed. Recovery of heme-iron increased with increasing Hgb concentration, whereas rejection of peptide and separation effciency expressed by HP ratio (heme-iron/peptide) did not show significant differences. HP ratio increased with increases in the degree of hydrolysis of Hgb and $KH_2PO_4$ concentrations of dialysis solution. Recovery of heme-iron decreased with increase in the pH of dialysis solution due to wash-out of heme-iron across the dialysis membrane caused by increase in solubility of heme-iron. Rejections of peptide were 74.5 and 87.5% (2 and 5 kDa of cut off size, respectively), whereas recovery of heme-iron decreased from 86.5 (2 kDa) to 63.1% (25 kDa). Amounts of heme-iron and peptide of dried heme-iron product were 21.7 and 77.0%, and HP ratio and production yield were 28.2 and 6.5%, respectively.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.84-91
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• 2012

Two staphylococcal lipases were obtained from Staphylococcus epidermidis S2 and Staphylococcus aureus S11 isolated from sebaceous areas on the skin of the human face. The molecular mass of both enzymes was estimated to be 45 kDa by SDS-PAGE. S2 lipase displayed its highest activity in the hydrolysis of olive oil at $32^{\circ}C$ and pH 8, whereas S11 lipase showed optimal activity at $31^{\circ}C$ and pH 8.5. The S2 lipase showed the property of cold-adaptation, with activation energy of 6.52 kcal/mol. In contrast, S11 lipase's activation energy, at 21 kcal/mol, was more characteristic of mesophilic lipases. S2 lipase was stable up to $45^{\circ}C$ and within the pH range from 5 to 9, whereas S11 lipase was stable up to $50^{\circ}C$ and from pH 6 to 10. Both enzymes had high activity against tributyrin, waste soybean oil, and fish oil. Sequence analysis of the S2 lipase gene showed an open reading frame of 2,067 bp encoding a signal peptide (35 aa), a pro-peptide (267 aa), and a mature enzyme (386 aa); the S11 lipase gene, at 2,076 bp, also encoded a signal peptide (37 aa), pro-peptide (255 aa), and mature enzyme (399 aa). The two enzymes maintained amino acid sequence identity of 98-99% with other similar staphylococcal lipases. Their microbial origins and biochemical properties may make these staphylococcal lipases isolated from facial sebaceous skin suitable for use as catalysts in the cosmetic, medicinal, food, or detergent industries.

• KSBB Journal
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• v.13 no.1
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• pp.114-118
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• 1998

The effects of fibron solubilization conditions on molecular weight distribution of enzymatically-hydrolyzed silk peptides were investigated. The weight-averaged molecular weights of silk proteins prepared by solubilization with calcium chloride, ethylenediamine and sulfuric acid were 41600, 3308, and 1268 dalton, respectively. Silk peptides in the average molecular weight range of 600-1200 dalton were obtained by protease treatment from solubilized silk fibroin. After the acid hydrolysis of silk protein using hydrochloric acid for 24 hr, silk protein was hydrolyzed to peptides whose average molecular weight and free amino acid content were 145 dalton and 80%, respectively. It was possible to control molecular weight distribution of silk peptides by the combination of solubilization and hydrolysis methods. Among the various treatment methods, acid solubilization followed by protease treatment had an advantage of molecular weight control for the peptide production.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.10
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• pp.1460-1468
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• 2003

An experiment was conducted to quantify the flow of soluble non-ammonia nitrogen (SNAN) in the liquid phase of ruminal (RD) and omasal digesta (OD), and to investigate diurnal pattern in SNAN flow in OD. Five ruminally cannulated Finnish-Ayrshire dairy cows in a $5{\times}5$ Latin square design consumed a basal diet of grass silage and barley grain, and that supplemented with four protein feeds (kg/d DM basis) as follows: skimmed milk powder (2.1), wet distiller' solubles (3.0), untreated rapeseed meal (2.1) and treated rapeseed meal (2.1). Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 1.0 h interval during a 12 h feeding cycle. Both RD and OD were acidified, centrifuged to remove microbes and precipitated with trichloroacetic acid followed by centrifugation. The SNAN fractions (free amino acid (AA), peptide and soluble protein) in RD and OD were assessed using ninhydrin assay. Free AA, peptide and soluble protein averaged 60.0, 89.4 and 2.1 g/d, respectively, for RD, and 81.8, 121.5 and 2.5 g/d, respectively, for OD. Although free AA flow was relatively high, mean peptide flow was quantitatively the most important fraction of SNAN, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis. Diurnal pattern in flow of peptide including free AA in OD during a 12 h feeding cycle peaked 1 h post-feeding, decreased by 3 h post-feeding and was relatively constant thereafter. Protein supplementation showed higher flow of peptide including free AA immediately after feeding compared with no supplemented diet. There were no differences among protein supplements in diurnal pattern in flow of peptide including free AA in OD.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.33-42
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• 2021

Due to the potential of antioxidants to scavenge free radicals in human body, it is important to be able to prepare antioxidant peptides that meet the industrial requirements for cosmetics and food. Here, we determined in vivo/in vitro activities of antioxidant peptide from P. fucata (PFAOP) prepared by bio-fermentation method. The antioxidant property test results showed the DPPH, hydroxyl, superoxide radical-scavenging, and cellular antioxidant activity. EC50 values of PFAOPs were 0.018 ± 0.005, 0.126 ± 0.008, 0.168 ± 0.005, and 0.105 ± 0.005 mg/ml, respectively, exhibiting higher antioxidant activities than glutathione (p < 0.05). Moreover, anti-proliferation and cytotoxicity activity results illustrated PFAOP has a potent anti-proliferative activity against HepG2, Caco-2, and MCF-7 carcinoma cells with no cytotoxicity. Moreover, the protocols we developed in this work demonstrated several excellent advantages in PFAOP preparation compared to enzymatic hydrolysis or chemical synthesis methods and provide a theoretical foundation for higher-value application of marine-derived functional peptides.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.509-513
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• 1999

The molecular size reduction and the formation of bitterness during a tryptic hydrolysis of soybean 11S glycinin were determined by using quantitative analysis and organoleptic evaluation. The 11S glycinin of 90% purity was prepared by cryoprecipitation and Con A Sepharose 4B affinity chromatography, and hydrolyzed with trypsin in a pH-stat reactor for 4 h. Bitterness was formed within 1 h of hydrolysis, and then slowly increased up to $3.5\times10^{-5}$ M quinine-HCl equivalent. The extent of hydrolysis (DH) was 7% at 1 h and increased up to 12% by the end of the reaction. The -amino nitrogen content increased from an initial 0.7 mM to 7 mM at the end of the period. The SDS-PAGE analysis showed that the acidic subunit of 11S glycinin was mostly hydrolyzed. The GP-HPLC analysis indicated that the bitterness was mainly contributed by the peptide fractions of molecular weights of 360-2,100 Da.

• Fisheries and Aquatic Sciences
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• v.25 no.7
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• pp.357-379
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• 2022

Increased fisheries products have raised by-products that are discarded due to low economic value. In addition, marine by-products are still rich in protein and nutritional value that have biological activities and give benefits to human health. Meanwhile, there is raised pressure for sustainability practices in marine industries to reduce waste and minimize the detrimental effect on the environment. Thus, valorization by-products through bioactive peptide mining are crucial. This review focus on various ways to obtain bioactive peptides from marine by-products through protein hydrolysis, for instance chemical hydrolysis (acid and based), biochemical hydrolysis (autolysis and enzymatic hydrolysis), microbial fermentation, and subcritical water hydrolysis. Nevertheless, these processes have benefits and drawbacks which need to be considered. This review also addresses various biological activities that are favorable in pharmaceutical industries, including antioxidant, antihypertensive, anticancer, anti-obesity, and other beneficial bioactivities. In addition, some potential marine resources of Indonesia for the marine biopeptide from their by-product or undesired marine commodities would be addressed as well.