• Journal of Life Science
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• v.21 no.2
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• pp.309-315
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• 2011

Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.961-967
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• 2005

Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.

• Korean Journal of Food Science and Technology
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• v.30 no.4
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• pp.908-915
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• 1998

By using Korean native soybean, traditional meju was prepared in Chuncheon, Kangweondo according to the traditional process. Analysis of physico-chemical, enzymatic and microbiological changes during meju fermentation were carried out in order to obtain a basic information for industrial scale production of meju. The enviroments for natural meju fermentation were $10{\sim}15^{\circ}C$ and $60{\sim}70%{\;}RH$. Moisture content decreased from 59% to 11% (exterior section) and 19% (interior section). the pH of meju rapidly increased up to 8.5 at $33^{rd}{\;}day$ of fermentation and thereafter decreased down to 7.9 at $70^{th}{\;}day$ of fermentation. Souble protein content was 1.47% at initial stage and increased up to $6.31{\sim}7.34%$ at $33^{rd}{\;}day$ of fermentation. Amino nitrogen content was $460{\sim}770{\;}mg%$ at $70^{th}{\;}day$ of fermentation. the color of meju became gradually black and decreased in redness and yellowness. During the process, protease and lipase seemed to play an important role in the digestion of soy protein and fat. Acidic protease activity increased up to $135.9{\sim}152.4{\;}unit/g$ at $33^{rd}{\;}day$ of fermentation and were $181.3{\sim}272.6{\;}unit/g$ at $70^{th}{\;}day$ of fermentation. Lipase activity increased up to 6 unit/g (interior section) and 15 unit/g (exterior section) at $70^{th}{\;}day$ of fermentation. the viable cell count of meju was at the level of $10^8{\;}CFU/g$ during the overall fermentation period. Aerobic halophilic count was $1.51{\times}10^7{\;}CFU/g$ at initial stage and maintained $10^8{\;}CFU/g$ level during the process. Initial anaerobic cell count was $2.0^9{\times}10^4{\;}CFU/g$ and increased up to $10^5{\;}CFU/g$ level at 47 days. Yeast and mold counts were $10^4{\sim}10^5{\;}CFU/g$ for the fermentation period.

• Korean Journal of Food Science and Technology
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• v.45 no.1
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• pp.25-33
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• 2013

Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.4
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• pp.481-487
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• 1999

The optimal conditions of enzymatic hydrolysis for preparation of rapid salted and fermented anchovy sauce (SFAS) using various pretenses such as trypsin, chymotrypsin, crude enzyme from squid liver and viscera, Alcalase, Neutrase and Protamex were studied. SFAS prepared with squid viscera had higher level of VBN (173.6 mg/100 g) when stored for 70 days than other samples, and peroxide values were almost equal among all samples during fermentation period. Total amino acids and nonprotein nitrogenous compounds remarkably increased as SFAS treated with Alcalase or Protamex which exhibited higher the hydrolysis rate of $57\%$ at 60 day than others. The optimal pHs of trypsin, chymotryosin, Alcalase, Neutrase and Protamex on anchovy actomyosin were 7.5, 6.5, 6.5, 7.0 and 5.0, respectively. Optimal temperatures of trypsin, chymotryosin, Alcalase and Neutrase were 55, 45, 60 and $55^{\circ}C$, respectively. Otherwise, Protamex activity increased as temperature increased from 20 to $70^{\circ}C$. Protamex had higher $K_m$ (3.545) and $V_{max}$ value (2.688) than others. Protamex affected less by NaCl had $52.5\%$ activity at the fermentation condition of $20^{\circ}C$ and $25\%$ NaCl. Protamex appeared to be very effective for the hydrolysis of crude actomyosin from ancnovy.

• Korean Journal of Food Science and Technology
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• v.51 no.5
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• pp.480-485
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• 2019

The present study was conducted to compare antioxidant capacities of protein hydrolysates from four different edible insects (Protaetia brevitarsis larvae, Allomyrina dichotoma larvae, Gryllus bimaculatus imago, and Tenebrio molitor larvae) which have recently been registered as food varieties in Korea. Protein hydrolysates were prepared from each insect using enzymatic hydrolysis using alcalase, and were then separated into a fraction containing ${\leq}3kDa$. According to $RC_{50}$ values and trolox equivalent antioxidant capacity results obtained from five different antioxidant analyses, the Gryllus bimaculatus (GB) hydrolysate showed relatively high levels of antioxidant capacity and, in particular, the GB hydrolysate showed considerably strong antioxidant activities in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and in ferric reducing antioxidant power (FRAP) assays. The GB hydrolysate also showed the strongest inhibitory effect on peroxidation of linoleic acid, and its rate of inhibition at $100{\mu}g/mL$ on day 3 of treatment was 60.26%. These results suggest that protein hydrolysates from edible insects including GB represent potential sources of natural antioxidants.

• Food Science and Preservation
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• v.19 no.4
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• pp.470-476
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• 2012

Enzymatic action and microbial growth degrade the quality of fresh-cut lettuce. Browning, a bad smell, and softening during storage are the major forms of quality deterioration. Health-oriented consumers tend to avoid foods treated with chemicals to maintain their freshness. This study was conducted to evaluate the change in the quality of fresh-cut lettuce with combined low-temperature blanching (LB) and ultrasonication (US). The optimum condition was selected using the response surface methodology (RSM), through a regression analysis with the following independent variables; the ultrasonication time (X1), blanching temperature (X2), blanching time (X3), and dependent variable; ${\Delta}E$ value (y). It was found that the condition with the lowest ${\Delta}E$ value occurred with combined 90s US and $45^{\circ}C$ 90s LB (US+LB). The combined treatment group (US+LB) was stored at $10^{\circ}C$ for 9 days with the control group and each single-treatment group, with low-temperature blanching and ultrasonication. Overall, the US+LB group had a significantly high $L^*$ value, which indicates significantly low $a^*$, $b^*$, ${\Delta}E$, browning index, PPO, and POD activity values, and a low total bacteria count (p < 0.05). The US+LB group also had the highest sensory score (except for aroma and texture; p > 0.05).

• Korean Journal of Food Science and Technology
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• v.53 no.4
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• pp.382-390
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• 2021

In this study, solubilized yuzu pulp powder (EYP) was produced using enzyme treated yuzu pulp powder (YP) and used to manufacture yuzu granules (0-20% EYP content). The physicochemical, product stability, and functional properties of Yuzu granules were compared among five enzyme treatments. Among the five treatments, CL had the highest YP solubilization yield (48.68%). Microstructural observation of EYP using FE-SEM revealed that its surface became irregular and porous after enzymatic treatment. Compared to YP, EYP had 2 times lower insoluble dietary fibers and 3 times lower hemicellulose and cellulose content. Among the yuzu granules, IV (yuzu granules with 15% EYP) had an excellent water and oil holding capacity and flowability. IV granule had the highest narirutin and hesperidin content of 3.4 mg and 2.2 mg/g DW, respectively and the highest antioxidant (68.4%) and tyrosinase inhibitory activities (82.5%). Therefore, EYP or granule with EYP can be used as a functional component in food industry or pharmaceutical field.

• Journal of the Korean Applied Science and Technology
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• v.38 no.3
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• pp.824-831
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• 2021

1, 2-Octanediol galactoside (OD-gal) has been synthesized from 1, 2-octanediol (OD), as a safer cosmetic preservative, using recombinant Escherichia coli β-galactosidase (β-gal). To confirm the molecular structure of synthesized OD-gal, mass spectrometry and NMR (¹H- and ¹³C-) spectroscopy of OD-gal were carried out. In the reaction mixture, a sodium adduct ion of OD-gal (m/z=331.1732) was identified using mass spectrometry analysis. In addition, ¹H NMR spectrum of OD-gal showed multiple peaks corresponding to the galactosyl group, which is evidence of galactosylation on OD. Downfield proton peaks at δ_H 4.39 ppm and multiple peaks from δ_H 3.98~3.55 ppm were indicative of galactosylation on OD. Up field proton peaks at δ_H 1.52~1.26 ppm and 0.89 ppm showed the presence of CH₂ and CH₃ protons of OD. ¹³C NMR spectrum revealed the presence of 24 carbons suggestive of α- and β-anomers of OD-gal. Among 14 carbon peaks from each anomer, the 4 peaks at δ_C 31.4, 29.0, 22.3, and 13.7 ppm were assigned to be overlapped showing only 24 peaks out of a total of 28 peaks. The mass value from mass spectrometry analysis of OD-gal, and ¹H and ¹³C NMR spectral data were in good agreement with the expecting structure of OD-gal. Finally, we identified a galactose molecule from the hydrolysate of OD-gal using β-gal. We are expecting that through future study it will eventually be able to develop a safe cosmetic preservative.

• Journal of Life Science
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• v.31 no.8
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• pp.719-728
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• 2021

Melanin pigments are abundantly distributed in mammalian skin, hair, eyes, and nervous system. Under normal physiological conditions, melanin protects the skin against various environmental stresses and acts as a physiological redox buffer to maintain homeostasis. However, abnormal melanin accumulation results in various hyperpigmentation conditions, such as chloasma, freckles, senile lentigo, and inflammatory pigmentation. Tyrosinase, a copper-containing enzyme, plays an important role in the regulation of the melanin pigment biosynthetic pathway. Although several whitening agents based on tyrosinase inhibition have been developed, their side effects, such as allergies, DNA damage, mutagenesis, and cytotoxicity of melanocytes, limit their applications. In this study, we synthesized 4-chromanone derivatives (MHY compounds) and investigated their ability to inhibit tyrosinase activity. Of these compounds, (E)-3-(4-hydroxybenzylidene)chroman-4-one (MHY1294) more potently inhibited the enzymatic activity of tyrosinase (IC₅₀ = 5.1±0.86 μM) than kojic acid (14.3±1.43 μM), a representative tyrosinase inhibitor. In addition, MHY1294 showed competitive inhibitory action at the catalytic site of tyrosinase and had greater binding affinity at this site than kojic acid. Furthermore, MHY1294 effectively inhibited α-melanocyte stimulating hormone (α-MSH)-induced melanin synthesis and intracellular tyrosinase activity in B16F10 melanoma cells. The results of the present study indicate that MHY1294 may be considered as a candidate pharmacological agent and cosmetic whitening ingredient.