• Korean Journal of Food Science and Technology
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• v.41 no.2
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• pp.179-185
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• 2009

Soybean grits, fortified with various bioactive components, were produced by solid-state fermentation using Bacillus subtilis HA. ${\alpha}$-Amylase activity gradually increased during fermentation over 5 days. Fibrinolytic and protease activities were highest in the soybean grits fermented for 7 days. The grits fermented for 5 days also showed the highest tyrosine content, indicating a higher peptide content. Peptides of low molecular weight (below 3,000 daltons) and browning pigments increased with increasing fermentation time. The fermented soybean grits showed higher contents of total phenolic compounds, to approximately 18 mg/g. DPPH free radical scavenging effects were higher in the soybean grits fermented for 3 days. Also, ABTS radical scavenging effects were greater in the fermented grits compared to the unfermented grits. Overall, the soybean grits fermented by solid-state fermentation for 5 days showed enhanced production of bioactive compounds and greater antioxidant properties.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1175-1182
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• 2021

We investigated the effect of bovine lactoferricin (Lfcin-B), a peptide derived from bovine lactoferrin, on activation of intestinal epithelial cells in IEC-6 intestinal cell, and protection against in vivo rotavirus (RV) infection. Treatment with Lfcin-B significantly enhanced the growth of IEC-6 cells and increased their capacity for attachment and spreading in culture plates. Also, Lfcin-B synergistically augmented the binding of IEC-6 cells to laminin, a component of the extracellular matrix (ECM). In the analysis of the intracellular mechanism related to Lfcin-B-induced activation of IEC-6 cells, this peptide upregulated tyrosine-dependent phosphorylation of focal adhesion kinase (FAK) and paxillin, which are intracellular proteins associated with cell adhesion, spreading, and signal transduction during cell activation. An experiment using synthetic peptides with various sequences of amino acids revealed that a sequence of 9 amino acids (FKCRRWQWR) corresponding to 17-25 of the N-terminus of Lfcin-B is responsible for the epithelial cell activation. In an in vivo experiment, treatment with Lfcin-B one day before RV infection effectively prevented RV-induced diarrhea and significantly reduced RV titers in the bowels of infected mice. These results suggest that Lfcin-B plays meaningful roles in the maintenance and repair of intestinal mucosal tissues, as well as in protecting against intestinal infection by RV. Collectively, Lfcin-B is a promising candidate with potential applications in drugs or functional foods beneficial for intestinal health and mucosal immunity.

• Korean Journal of Food Science and Technology
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• v.46 no.2
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• pp.165-172
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• 2014

Mixed fermentation of cow milk was performed by sequential co-cultures with Bacillus subtilis and Lactococcus lactis. After a first fermentation step with B. subtilis for 6 h, the number of viable cells increased to $2.5{\times}10^8$ CFU/mL. The second fermentation step with L. lactis resulted in increased viable cells $1.09{\times}10^{10}$ CFU/mL for 3 days and increased acidity. However, the number of viable B. subtilis cells was decreased greatly to $5{\times}10^1$ CFU/mL following fermentation with L. lactis. Milk proteins were markedly hydrolyzed by the first fermentation after 2 h, and the second fermentation induced curd formation in milk. However, after 4 h, the first fermentation resulted in higher whey separation and 80 mg% tyrosine content. Gamma-aminobutyric acid (GABA) production was dependent upon the degree of protein hydrolysis by first fermentation. Second fermentation resulted in 0.14% GABA. The milk fermented by B. subtilis indicated the rough surface of yogurt depended upon the degree of protein hydrolysis. In conclusion, set-type yogurt was efficiently produced by co-culturing of milk, and fortifying with peptides, GABA, and probiotics.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.9
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• pp.211-226
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• 2019

Pig CD45, the leukocyte common antigen, is encoded by the PTPRC gene and CD45 is a T cell-type specific tyrosine phosphatase with alternative splicing of its exons. The CD45 is a coordinated regulator of T cell antigen receptor (TCR) signal transduction achieved by dephosphorylating the phosphotyrosine of its substances, including $CD3{\zeta}$ chain of TCR, Lck, Fyn, and Zap-70 kinase. A dysregulation of CD45 is associated with a multitude of immune disease and has been a target for immuno-drug discovery. To characterize its key structural features with the effects of regulating TCR signaling, this study predicted the unknown structure of pig CD45RO (the smallest isoform) and the complex structure bound to the ITAM (REEpYDV) of $CD3{\zeta}$ chain via homology modeling and docking the peptide, based on the known human CD45 structures. These features were integrated into the structural plasticity of extracellular domains and functional KNRY and PTP signature motifs (the role of a narrow entrance into ITAM binding site) of the tyrosine phosphatase domains in a cytoplasmic region from pig CD45RO. This contributes to the selective recognition of phosphotyrosine from its substrates by adjusting the structural stability and binding affinity of the complex. The characterized features of pigCD45RO can be applied in virtual screening of the T-cell specific immunomodulator.

• Journal of Microbiology and Biotechnology
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• v.13 no.4
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• pp.483-486
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• 2003

3'-[S-Methyl-cysteinyl]-3'-amino-3'-deoxy-N,N- dimethyl adenosine, cystocin, is a biosynthesized antibiotic material newly identified from Streptomyces sp. GCA0001. Its structure was found to be similar to puromycin, where the terminal tyrosine is replaced by a methyl cysteine. NMR data prove that the 3-ammo ribose is connected to dimethylaminopurine through the anomeric carbon at 1'-carbon. The methyl cysteinyl unit is connected to the amino unit of ribose by peptide bond. The verification of the structure was performed by comparing the puromycin nucleosides resulted from the hydrolysis of cystocin and puromycin, respectively. Antibiotic activity of cystocin against Streptococcus was found to be two times more potent than that of puromycin.

• Animal cells and systems
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• v.2 no.2
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• pp.243-248
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• 1998

We have partially sequenced glutaminyl cyclases from several mammalian and one avian species and found that the two cysteine residues of the human glutaminyl cyclase are completely conserved. The mammalian glutaminyl cyclase has been reported to possess reactive thiols (Busby, Jr, et aI., 1987, J BioI Chern 262, 8532-8536). Mutagenesis of these cysteine residues, however, resulted in only a slight decrease in enzyme activity. Likewise, the recombinant human enzyme was completely resistant to attempted chemical modification of the putative reactive thiols. Although the human glutaminyl cyclase did not appear to have reactive thiols, it was sensitive to diethylpyrocarbonate and acetylimidazole, indicating the presence of functionally important histidine and tyrosine residues which could act as acid/base catalysts. Almost identical deuterium solvent isotope effect (1.2 vs 1.3) upon the reaction by the human and papaya enzymes, respectively, provides an evidence both animal and plant glutaminyl cyclases catalyze pyroglutamyl-peptide formation by intramolecular cyclization.

• Food Science of Animal Resources
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• v.42 no.1
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• pp.46-60
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• 2022

In this study, angiotensin-I-converting enzyme inhibitory (ACEI) activity was evaluated in fermented goat milk fermented by lactic acid bacteria (LAB) from fermented foods and breast milk. Furthermore, the potential for ACEI peptides was identified in fermented goat milk with the highest ACEI activity. The proteolytic specificity of LAB was also evaluated. The 2% isolate was inoculated into reconstituted goat milk (11%, w/v), then incubated at 37℃ until pH 4.6 was reached. The supernatant produced by centrifugation was analyzed for ACEI activity and total peptide. Viable cell counts of LAB and titratable acidity were also evaluated after fermentation. Peptide identification was carried out using nano liquid chromatography mass spectrometry (LC-MS/MS), and potential as an ACEI peptide was carried out based on a literature review. The result revealed that ACEI activity was produced in all samples (20.44%-60.33%). Fermented goat milk of Lc. lactis ssp. lactis BD17 produced the highest ACEI activity (60.33%; IC₅₀ 0.297±0.10 mg/mL) after 48 h incubation, viable cell counts >8 Log CFU/mL, and peptide content of 4.037±0.27/mL. A total of 261 peptides were released, predominantly derived from casein (93%). The proteolytic specificity of Lc. lactis ssp. lactis BD17 through cleavage on the amino acid tyrosine, leucine, glutamic acid, and proline. A total of 21 peptides were identified as ACEI peptides. This study showed that one of the isolates from fermented food, namely Lc. lactis ssp. lactis BD17, has the potential as a starter culture for the production of fermented goat milk which has functional properties as a source of antihypertensive peptides.

• Journal of the Korean Chemical Society
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• v.5 no.1
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• pp.33-37
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• 1961

We have shown that carboxy-peptidase destroys the biological activity of angiotensin octa-and deca-peptides. Since Proline occurs as the seventh amino acid from the amino end of the chain and since carboxypeptidase does not cleave proline from a peptid chain, it is evident that the heptapeptid H.asp-arg-val-tyr-ileu-his-pro.OH is formed by this hydrolysis. This peptide must then be biologically inactive. In order to determine whether the phenyl group of the C-terminal amino acid was the necessary requirement for biological activity of the octapeptide, $ala^8$ angiotensin octapeptide(amino acids of peptides numbered from amino end) was synthesized. For this synthesis the four dipeptides were prepared: carbobenzoxy-L-prolyl-L-alanine-P-nitrobenzyl-ester, m.p. $134-135^{\circ}C,$ carbobenzoxy-L-isoleucyl-imidazole benzyl-L-histidine methyl ester, m.p. $114-116^{\circ}C,$ carbobenzoxy-L-valyl-L-tyrosine hydrazide and carbobenzoxy B-benzyl-L-aspartyl-nitro-L-arginine. The first three dipeptides were obtained as crystalline compounds. Imidazole-benzyl-L-histidine was used in the hope that it would block the histidine imidazole against side reactions in steps subsequent to the formation of the C-terminal tetrapeptide. Also, it was through that the imidazole benzylated peptides would be easier to crystallize. This, however, was not the case. The tetrapeptide, carbobenzoxy-L-isoleucyl-L-im, benzyl-histidyl, L-prolyl-L-alanine-nitrobenzyl ester was not obtained in a crystalline form. Neither could the mono-or dihydrobromide of the tetrapeptide free base be induced to crystallize. Carbobenzoxy-L-valyl-L-tyrosine azide was condensed with the tetrapeptide free base to yield the protected hexapeptide; carbobenzoxy-L-valyl-L-tyrosyl-L-isoleucyl-L-im, benzyl, histidyl-L-Prolyl-L-alanine-nitrobenzyl ester. Upon removal of the carbobenzoxy group with hydrogen bromide in acetic acid an amorphous free base hexapeptide ester was obtained. This compound gave the correct C, H, N analysis and contained the six amino acids in the correct ratio. The octapeptide was obtained by condensing this hexapeptide with carbobenzoxy-B-benzyl-L-aspartyl-nitro, L-arginine using the mixed anhydride method of condensation. This amorphous product was proven to be homogenous by chromatography in two solvent systems and upon hydrolysis yielded the eight amino acids in correct ratio. The five protecting groups were removed from the octapeptide by hydrogenolysis over palladium black catalyst. Biological assay of the free peptide indicated that it possessed less than 0.1 per cent of both pressor and oxytocic activity of the phenylalanine8 angiotensin. This suggests that the phenyl group is a point of attachment between angiotensin and its biological receptor site.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3053-3059
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• 2012

Human epidermal growth factor receptor 2 (HER2) is a member of the epidermal growth factor receptor family of receptor tyrosine kinases that plays important roles in all processes of cell development. Their overexpression is related to many cancers, including examples in the breast, ovaries and stomach. Anticancer therapies targeting the HER2 receptor have shown promise, and monoclonal antibodies against subdomains II and IV of the HER2 extra-cellular domain (ECD), Pertuzumab and Herceptin, are currently used in treatments for some types of breast cancers. Since anti HER2 antibodies targeting distinct epitopes have different biological effects on cancer cells; in this research linear and conformational B cell epitopes of HER2 ECD, subdomain III, were identified by bioinformatics analyses using a combination of linear B cell epitope prediction web servers such as ABCpred, BCPREDs, Bepired, Bcepred and Elliprro. Then, Discotope, CBtope and SUPERFICIAL software tools were employed for conformational B cell epitope prediction. In contrast to previously reported epitopes of HER2 ECD we predicted conformational B cell epitopes $P1_C$: 378-393 (PESFDGDPASNTAPLQ) and $P2_C$: 500-510 (PEDECVGEGLA) by the integrated strategy and P4: PESFDGD-X-TAPLQ; P5: PESFDGDP X TAPLQ; P6: ESFDGDP X NTAPLQP; P7: PESFDGDP-X-NTAPLQ; P8: ESFDG-XX-TAPLQPEQL and P9: ESFDGDP-X-NTAPLQP by SUPERFICIAL software. These epitopes could be further used as peptide antigens to actively immune mice for development of new monoclonal antibodies and peptide cancer vaccines that target different epitopes or structural domains of HER2 ECD.