• Applied Chemistry for Engineering
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• 제31권1호
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• pp.67-72
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• 2020

Biomaterials are frequently functionalized with Arg-Gly-Asp (RGD) peptides to provide cell adhesion sites. In this study, RGD peptides were enzymatically coupled on to the surface of fibroin microspheres. Papain exhibited a strong preference for dansyl phenylalanine for the peptide formation with fibroin microspheres. Thus, RGD1 peptide was designed to carry cysteine to both sides of the sequence, glycine as a spacer and two residues of phenylalanine at the C-terminal (CRGDCGFF). The enzymatic modification facilitated by an increasing amount of substrate and by the presence of organic solvent, dimethylsulfoxide at 25% (v/v). Microspheres coupled with RGD1, showed a significantly different precipitation property and an increased apparent volume, possibly due to the steric hindrance of RGD peptides on the surface. Transmission electron microscopy also confirmed the presence of cysteine residues in RGD1 coupled on the surface of microspheres stained with gold nanoparticles. RGD1-microspheres significantly facilitated the growth of murine fibroblast 3T3 cells even under non-adhesion culture conditions.

• Food Science of Animal Resources
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• 제35권4호
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• pp.533-540
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• 2015

The aim of this study was to investigate the effects of aqueous extract from Sarcodon aspratus on tenderization of the bovine longissimus dorsi muscles in comparison with commercial proteolytic enzymes. Furthermore, meat quality and muscle protein degradation were examined. We marinated meat with 2% Sarcodon aspratus extract, 2% kiwi extract, and 0.2% papain. Beef chunks (3×3×3 cm³) were marinated with distilled water (control), Sarcodon aspratus extract (T1), kiwi extract (T2) or papain (T3) for 48 h at 4℃. There were no significant differences in muscle pH and lightness between control and treated samples. T1 had the lowest redness (p<0.01), and higher cooking loss and water holding capacity than control and T2 (p<0.05). T1 and T3 exhibited lower shear force values than control (p<0.05). Total protein solubility did not differ significantly between T1 and control, but T1 had less myofibrillar protein solubility than control and T2 (p<0.001). The degradation of myosin heavy chain in T1 and T3 was observed. This degradation of myofibrillar protein suggests that Sarcodon aspratus extract could influence tenderization. These results show that aqueous extract of Sarcodon aspratus extract actively affect the tenderness of the bovine longissimus dorsi muscle.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.81-85
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• 2000

An elastase inhibitor, SMFEI02, was isolated from culture broth of Streptomyces lavendulae SMF11. The inhibitor was purified by ultrafiltration followed by XAD-7 column and Dowex-1 anion-exchange chromatographies, and preparative HPLC. The molecular formula was determined to be $C_{14}H_{16}N_2O_2$ (MW244) by HRFAB-MS analysis. The inhibitor was identified to be a diketopiperazine cyclo(S-Phe-S-Pro) by the optical rotation value and MNR spectral data, and showed inhibitory activities for trypsin, chymotrypsin, cathepsin B, and papain as well as elastase with the Ki values ranging from 1.78mM to $2.86{\;}\mu\textrm{m}$. The inhibition showed a competitive mode for elastase, chymotrypsin, and cathepsin B, whereas it showed a noncompetitive mode for trypsin and papain.

• Food Science of Animal Resources
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• 제40권2호
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• pp.172-182
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• 2020

In the present study, we determined the degree of hydrolysis (DH) of porcine blood plasma proteins, albumin, and globulin hydrolyzed by six proteases (alcalase, neutrase, flavourzyme, protamex, trypsin, and papain) for various reaction times. Moreover, antimicrobial activities of hydrolysates against five pathogenic microorganisms (Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium, Escherichia coli, and Shigella flexneri) were investigated. Alcalase, trypsin, and papain hydrolysates of the three porcine blood proteins showed higher DH values than hydrolysates produced by the other three proteases. DH of the three porcine blood proteins hydrolyzed by the six proteases failed to increase after 2 h of hydrolysis. In antimicrobial tests, hydrolysates (hydrolysis time of 2 h) showed antibacterial activity only against B. cereus. Albumin hydrolysates showed higher antimicrobial activity than globulin and plasma hydrolysates. Albumin hydrolysates obtained with flavourzyme, protamex, and trypsin showed higher antimicrobial activity than those obtained with the other three proteases.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.283-288
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• 2011

The jellyfish Nemopilema nomurai was hydrolyzed with papain and a novel dipeptide purified via ultrafiltration, gel filtration chromatography with Sephadex LH-20, and reverse phase chromatography using $C_{18}$ and $C_{12}$ columns. The IR, 1H NMR, 13C NMR, and MS spectrometer analyses showed that the dipeptide comprised tyrosine-isoleucine (Tyr-Ile). The $IC_{50}$ and $K_i$ values were $6.56{\pm}1.12$ and $3.10{\pm}0.28\;{\mu}M$, respectively, indicating competitive inhibition of angiotensin-I-converting enzyme (ACE). As a novel ACE-inhibitory active peptide, Tyr-Ile may have potential for use in antihypertensive therapy.

• Korean Journal of Food Science and Technology
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• 제5권3호
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• pp.174-177
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• 1973

Total and free tryptophan contents in the fore shank muscle of korean cow treated with proteolytic enzymes have been analyzed by the spectrophotometric method and the results are as follows: 1. By adding 0.01%, 0.05%, 0.1% of proteolytic enzymes, the total tryptophan and free tryptophan have incresed steadily 2. The rate of increase of the total tryptophan content was highest when 0.05% of the enzymes for the meat weight were added. 3. The change in free tryptophan incident to the addition of proteolytic enzymes was insignificant.

• IMMUNE NETWORK
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• 제13권6호
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• pp.249-256
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• 2013

How Th2 immunity develops in vivo remains obscure. Basophils have been considered key innate cells producing IL-4, a cytokine essential for Th2 immunity. Increasing evidence suggests that basophils are dispensable for the initiation of Th2 immunity. In this study, we revisited the role of basophils in Th2 immune responses induced by various types of adjuvants. Mice deficient in IL-3 or IL-3 receptor, in which basophil lymph node recruitment is completely abolished, fully developed wild type level Th2 CD4 T cell responses in response to parasite antigen or papain immunization. Similar finding was also observed in mice where basophils are inducibly ablated. Interestingly, IL-4-derived from non-T cells appeared to be critical for the generation of IL-4-producing CD4 T cells. Other Th2 promoting factors including IL-25 and thymic stromal lymphopoietin (TSLP) were dispensable. Therefore, our results suggest that IL-3- and basophil-independent in vivo Th2 immunity develops with the help of non-T cell-derived IL-4, offering an additional mechanism by which Th2 type immune responses arise in vivo.

• Journal of Microbiology and Biotechnology
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• 제16권4호
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• pp.524-530
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• 2006

Two protease inhibitors of 67 and 18 kDa, respectively, were purified from the eggs of glass fish, Liparis tanakai, by affinity chromatography and electro-elution method. The high molecular weight (HMW) protein was purified with a specific inhibitory activity, yield, and purity of 18.46 U/mg, 0.07%, and 131.86 fold, respectively, and was further characterized: Optimal temperature and pH for inhibitory activity of the HMW glassfish egg protease inhibitor were $40^{\circ}C$ and pH 6, respectively, and it was stable between $5^{\circ}C\;and\;50^{\circ}C$ in the pH range of 5-6 with maximal stability at pH 6. It was shown to be a competitive inhibitor against papain with an inhibition constant $(K_i)$ of 97.02 nM. Moreover, the 67 kDa protein inhibited cathepsin, a cysteine protease, more effectively than did an egg-white protease inhibitor. The HMW glassfish egg protease inhibitor was classified as a member of the family III (kininogen).

• Food Science and Biotechnology
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• 제17권4호
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• pp.873-877
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• 2008

Native soy protein isolate (SPI) was hydrolyzed with 4 different proteolytic enzymes, including bromelain, papain, Neutrase, and Flavourzyme. SPI hydrolysates with the degree of hydrolysis (DH) in range of 6 to 15% were prepared by each enzyme. The angiotensin 1 converting enzyme (ACE) inhibitory and the antioxidant activities of the SPI hydrolysates, such as superoxide dismutase-like activity and inhibition of the linoleic acid autoxidation, were evaluated. Overall, as the DH increased, all evaluated bioactivities of the SPI hydrolysates significantly increased. The significantly highest ACE inhibitory and antioxidant activities were found in hydrolysates made with papain and bromelain, respectively. SPI hydrolysates by Flavourzyme showed the significantly lowest activity in all tested bioactivities. The results suggested that ACE inhibitory and antioxidant activities of SPI hydrolysates were determined by the DH and by the enzyme used.

• Journal of the Society of Cosmetic Scientists of Korea
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• 제26권1호
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• pp.107-124
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• 2000

A bacterial strain No. HB-5, which was capable of producing a pretense in the culture conditions, was isolated from the soil . The pretense was purified from cultural filtrate of Bacillus sp. HB-5 by membrane ultrafiltration and DEAE- cellulose chromatography, gel filtration on Sephadex G-100. The molecular weight was estimated to be 60k4a. The optimal pH and temperature for the activity of the purified pretense pH were 11 and 5$0^{\circ}C$ , respectively. The enzyme was stable within a pH range 8-12 and up to 6$0^{\circ}C$ . The enzyme activity was highly inhibited by PMSF at 1mM. The proteolytic actions of pretense and papain on human epidermis keratins which are major protein impurities on the skin, were compared. The bacterial pretense degraded more effectively than papain. Product containing 2% protease exhibited 21% increase on the skin coloration index. These results suggest that cosmetic product containing pretense produced by Bacillus sp. HB-5 could remove the adherent keratin layer and then make a softer skin.