• BMB Reports
• /
• v.55 no.4
• /
• pp.175-180
• /
• 2022

Peptides are gaining substantial attention as therapeutics for human diseases. However, they have limitations such as low bioavailability and poor pharmacokinetics. Periostin, a matricellular protein, can stimulate the repair of ischemic tissues by promoting angiogenesis. We have previously reported that a novel angiogenic peptide (amino acids 142-151) is responsible for the pro-angiogenic activity of periostin. To improve the in vivo delivery efficiency of periostin peptide (PP), we used proteins self-assembled into a hollow cage-like structure as a drug delivery nanoplatform in the present study. The periostin peptide was genetically inserted into lumazine synthase (isolated from Aquifex aeolicus) consisting of 60 identical subunits with an icosahedral capsid architecture. The periostin peptide-bearing lumazine synthase protein cage nanoparticle with 60 periostin peptides multivalently displayed was expressed in Escherichia coli and purified to homogeneity. Next, we examined angiogenic activities of this periostin peptide-bearing lumazine synthase protein cage nanoparticle. AaLS-periostin peptide (AaLS-PP), but not AaLS, promoted migration, proliferation, and tube formation of human endothelial colony-forming cells in vitro. Intramuscular injection of PP and AaLS-PP increased blood perfusion and attenuated severe limb loss in the ischemic hindlimb. However, AaLS did not increase blood perfusion or alleviate tissue necrosis. Moreover, in vivo administration of AaLS-PP, but not AaLS, stimulated angiogenesis in the ischemic hindlimb. These results suggest that AaLS is a highly useful nanoplatform for delivering pro-angiogenic peptides such as PP.

• Bulletin of the Korean Chemical Society
• /
• v.32 no.8
• /
• pp.2577-2583
• /
• 2011

We have designed a 20-residue hybrid peptide CA(1-8)-MA(1-12) (CAMA) incorporating residues 1-8 of cecropin A (CA) and residues 1-12 of magainin 2 (MA) with high bacterial cell selectivity. CAMA-P2 is an ${\alpha}$-helical antimicrobial peptide designed from a CAMA hybrid peptide and substitution of Gly-Ile-Gly hinge sequence of CAMA to Pro influences the flexibility at central part of CAMA. Based on structure-activity relationships of CAMA peptides, to investigate the effects of the total positive charges on antimicrobial activity of CAMA-P2, the $Ser^{14}{\rightarrow}$Lys analogue (CAMA-syn1) was synthesized. The role of tryptophan at C-terminal ${\alpha}$-helix on its antimicrobial activity as well as synergistic activity was also investigated using $Ser^{14}{\rightarrow}$Lys/$Phe^{18}{\rightarrow}$Trp analogue (CAMA-syn2). Also, we designed CAMA-syn3 by substitution of $Lys^{16}$ located opposite side of substituted $Lys^{14}$ of CAMA-syn1 with Leu residue, resulting in increase of hydrophobicity and amphipathicity of the peptide. All of CAMA-syn analogues showed good antimicrobial activities similar to those of CAMA and CAMA-P2. The CAMA-syn1 and CAMA-syn2 showed low hemolytic activity and cytotoxicity against human keratinocyte Haca-T cells while CAMA-syn3 showed hemolytic activity and cytotoxicity at its MIC value. We then investigated their abilities to act synergistically in combination with the antimicrobial flavonoids and synthetic compounds screened in our laboratory. The results showed that all peptides exhibited synergistic effects with dihydrobinetin, while only CAMA-syn2 exhibited synergistic effects with YKAs3001 against both S. aureus and MRSA, suggesting that Trp residue at C-terminus of CAMA-syn2 may facilitate the polar antibiotic flavonoids and synthetic compounds to permeabilize the membrane. This study will be useful for the development of new antibiotic peptides with potent antimicrobial and synergistic activity but without cytotoxicity.

• Korean Journal of Food Science and Technology
• /
• v.37 no.5
• /
• pp.827-832
• /
• 2005

Bacillus subtilis JM3 protease from naturally fermented anchovy sauce was partially purified in 40-60% ammonium sulfate fraction. To accelerate fermentation of anchovy sauce, 2 and 4% crude B. subtilis JM3 proteases were added to 6 month-ripened anchovy sauce, and their hydrolysis degrees and amino-nitrogen contents were investigated at different storage times. Low molecular weight (LMW) peptide was purified by ultrafiltration ana gel permeation chromatography from anchovy sauce manufactured with B, subtilis JM3 protease. Anchovy sauces with 2 and 4% proteases increased hydrolysis rate by 27 and 32%, respectively. Amino-nitrogen contents of anchovy sauces fermented with 2 and 4% proteases were twofold higher than that of control. Control showed five peptide peaks on Bio-Rad P2 gel permeation chromatography spectrum, whereas anchovy sauces with 2 and 4% B. subtilis JM3 proteases showed six and seven peaks, respectively. ACE inhibitory activity was highest in peak 6 (43.75%) of anchovy sauce with 2% protease, followed by peak 5 (34.82%) of control. DPPH radical-scavenging effect was higher than 50% in all samples. Cytotoxicity was highest in peak 3 (44.12%) of control, fellowed by peak 5 (42.04%) of anchovy sauce with 4% protease.

• Food Science of Animal Resources
• /
• v.35 no.6
• /
• pp.738-747
• /
• 2015

Angiotensin-converting enzyme (ACE) inhibitory activity was evaluated for the low-molecular-weight fraction (<3 kDa) obtained from milk fermentation by Bifidobacterium longum KACC91563. The ACE inhibitory activity in this fraction was 62.3%. The peptides generated from the <3 kDa fraction were identified by liquid chromatography-electrospray ionization quantitative time-of-flight mass spectrometry analysis. Of the 28 peptides identified, 11 and 16 were identified as β-casein (CN) and α_s1-CN, respectively. One peptide was identified as κ-CN. Three peptides, YQEPVLGPVRGPFPIIV, QEPVLGPVRGPFPIIV, and GPVRGPFPIIV, from β-CN corresponded to known antihypertensive peptides. We also found 15 peptides that were identified as potential antihypertensive peptides because they included a known antihypertensive peptide fragment. These peptides were as follows: RELEELNVPGEIVE (f1-14), YQEPVLGPVRGPFP (f193-206), EPVLGPVRGPFPIIV (f195-206), PVLGPVRGPFPIIV (f196-206), VLGPVRGPFPIIV (f197-206), and LGPVRGPFPIIV (f198-206) for β-CN; and APSFSDIPNPIGSENSEKTTMPLW (f176-199), SFSDIPNPIGSENSEKT- TMPLW (f178-199), FSDIPNPIGSENSEKTTMPLW (f179-199), SDIPNPIGSENSEKTTMPLW (f180-199), DIPNPIGSENSEKTTMPLW (f181-199), IPNPIGSENSEKTTMPLW (f182-199), PIGSENSEKTTMPLW (f185-199), IGSENSEKTTMPLW (f186-199), and SENSEKTTMPLW (f188-199) for α_s1-CN. From these results, B. longum could be used as a starter culture in combination with other lactic acid bacteria in the dairy industry, and/or these peptides could be used in functional food manufacturing as additives for the development of a product with beneficial effects for human health.

• Mass Spectrometry Letters
• /
• v.12 no.3
• /
• pp.81-84
• /
• 2021

Collagen, which accounts for one-third of human protein, is reduced due to human aging, and much attention is focused on making collagen into food to prevent such aging. Gel permeation chromatography with Reflective Index (RI) detection (GPC/RI) was chosen as the most suitable instrument to confirm molecular weight distribution, and we explored the use of this technique for analysis of collagen peptide molecular sizes and distributions. Data reliability was verified by matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometric analysis. The data were considered meaningful for comparative analysis of molecular weight distribution patterns.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.17 no.1
• /
• pp.137-141
• /
• 2008

We previously isolated and cloned a cDNA of abaecin from the Bombus ignitus. In an effort to produce a large amount of soluble abaecin at low cost, we successfully expressed the peptide in Escherichia coli that are highly sensitive to its mature form. For this, we fused the peptide encoding 39 amino acids of mature B. ignitus abaecin to the thioredoxin gene together with a C-terminal 6xHis tag. An enterokinase cleavage site was introduced between the 6xHis tag and mature abaecin to allow final release of the recombinant peptide. A high yield of 9.6 mg soluble fusion protein from 200 ml of bacterial culture was purified by $Ni^{2+}$-charged His-Bind resin affinity column, and 1.4 mg of pure active recombinant abaecin was readily obtained by enterokinase cleavage, followed by affinity chromatograph. The molecular mass of recombinant abaecin peptide was determined by Tricin-SDS-PAGE analysis. The recombinant abaecin exhibited antibacterial activity against Gram-negative bacteria.

• Journal of the Korean Applied Science and Technology
• /
• v.30 no.3
• /
• pp.560-566
• /
• 2013

In this research we extracted water-soluble collagen peptide from flatfish skin and compared it with commercially available collagen peptide extracted from Tilapia scale currently placed on the market in the aspect of physiochemical property. The physical property and nutritional components of FSCP appeared almost similarly to those of TSCP, and also in calorie, FSCP marking 3.82 Kcal showed no differences from TSCP marking 3.84 Kcal. As for forming amino acids, in aspartic acid, serine, histidine, tyrosine, methionine, FSCP had higher content than TSCP, but in OH-proline, proline and alanine FSCP had lower content than TSCP. Especially the content of essential amino acids of FSCP marked 22.74% with a higher content compared with 13.64% of TSCP. In the distribution of molecular weight FSCP with 1,000 Da showed comparatively low compared with TSCP, and in emulsion property and stability FSCP and TSCP showed similar excellent trend.

• Microbiology and Biotechnology Letters
• /
• v.46 no.4
• /
• pp.326-333
• /
• 2018

In this study, we identified methods to improve heat stability and skin permeability of functional protein biopolymers, such as growth factors, enzymes, and peptides. The biopolymers participate in cellular activation and catalytic functions in vivo. Therefore, when applied to cosmetics, their efficacies are expected to be helpful for skin care. However, they have disadvantages that include instability to heat and low skin permeability due to their high molecular weight. To overcome these problems, we searched for a composition that increases heat stability. Stability was improved using a polymeric humectant having a long polyethylene glycol length, compared with a mono-molecular structure humectant. Next, to enhance skin permeation, a permeation enhancing peptide was selected from a phage library. The permeation enhancing peptide can be commonly used to promote the permeation of growth factors, enzymes, and peptides. Screening was performed on the polymeric humectant formulation. One dominant peptide from the modified-screening method was identified. Furthermore, it was confirmed that the permeability of the peptide was better than that of the peptide developed through a screening system based on phosphate-buffered saline. The data indicate that the polymeric humectant formulation will be helpful for increasing the heat stability of protein ingredients and that skin permeability could be increased by a formulation-specific, penetration-enhancing peptide.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.58 no.1
• /
• pp.50-56
• /
• 2013

For accurate analysis of low molecular peptides using SELDI-TOF MS (surface enhanced laser desorption/ionization time of flight mass spectrometry), the optimized analytical conditions should be established for a specific biological sample. This study was conducted to optimize SELDI-TOF MS analytical conditions for profiling low molecular peptide below 10 kDa presented in sorghum seeds. Analytical conditions were as follows; (1) protein chips: CM10 (weak cation exchanger) and Q10 (strong anion exchanger), (2) dilution factors of binding buffer: 1/2, 1/5, 1/10, 1/20, 1/50, 1/100, and 1/200, (3) the stringency of Q10 binding buffer: 10 mM and 100 mM, and (4) protein extraction buffers: sodium borate, sodium borate + acetone, phenol, and TCA buffers. Optimum dilution factors were selected as 1/20 and 1/50 in both protein chips, CM10 and Q10. Low stringency of Q10 binding buffer (10mM) detected more peptide peaks than high stringency (100 mM). Selected protein extraction buffers of sorghum seed for SELDI-TOF MS analysis was the sodium borate buffer in the range of 2~10 kDa, while the phenol buffer was more suitable in the range of 10~20 kDa.

• Journal of Applied Biological Chemistry
• /
• v.62 no.1
• /
• pp.101-107
• /
• 2019

The objectives of this study were to evaluate the anti-aging effects and investigate the effect of simulated gastrointestinal (GI) digestion on the anti-aging properties and intestinal permeation of the potential fish collagen hydrolysates (FCH). Therefore, procollagen synthesis, matrix metalloproteinase-1 (MMP-1) production, and Caco-2 cell permeability were analyzed before and after in vitro digestion for FCHs, low-molecular weight fractions (<1 kDa), and high molecular weight fractions (>1 kDa). After being subjected to GI digestion, the level of MMP-1 inhibition was maintained, although the procollagen production was significantly (>20%) lower with all samples. Also, the digested FCHs and their <1 kDa fraction yielded 9.1 and 13.8% increased peptide transport, respectively, compared to undigested samples. Based on the effective intestinal permeation and high digestive enzyme stability, the <1 kDa fraction of FCHs is a potential bioactive material suitable for anti-aging applications in the food and cosmetics industries.