• 한국버섯학회지
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• 제14권4호
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• pp.162-167
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• 2016

Lactic acid bacteria (LAB) producing phenyllactic acid (PLA), which is known as antimicrobial compound, was isolated from button mushroom bed and the isolated LAB was identified to Lactobacillus casei by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. casei was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23 mM in CFS when L. casei was grown in MRS broth containing 5 mM phenylpyruvic acid as precursor for 16 h. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. casei with average growth inhibitions ranging from 34.58% to 65.15% (p < 0.005), in which R. solani was the most sensitive to 65.15% and followed by C. aculatum, and B. cinerea. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range of 0.35 mg mL-1 (2.11 mM) to 0.7 mg mL-1 (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens were not affected by the heating or protease treatment. However, pH modification in CFS to 6.5 resulted in an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS was caused by acidic compounds like PLA or organic acids rather than protein or peptide molecules.

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.440-446
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• 1989

Penicillium sp. CB-20의 polygalacturonase 생성을 위한 최적조건은 탄소원으로 펙틴을 사용하여 16 시간 배양시 최대 활성을 나타내었으며, Sephadex G-25, G-75 및 G-150을 사용한 gel filtration과 DEAE-cellulose, DEAE-Sephadex A-50에 의 한 ion exchange chromatography를 통하여 이 효소를 약 30배 가량 정제할 수 있었고, 수율은 2.31%였다. 정제효소는 polyacrylamide gel electrophoresis에 의하여 단일 밴드로 확인 되었으며, 분자량은 SDS PAGE에 의하여 21,000 정도로 측정되었다. 효소의 결정구조는 마름모꼴을 형성하고 있었으며 아미노산 조성은 17종류로써 glutamic acid, glycine, hlstidine의 함량이 비교적 많았다.

• 한국미생물·생명공학회지
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• 제43권2호
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• pp.91-96
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• 2015

Prion은 양의 scrapie, 소의 bovine spongiform encephalopathy와 사람의 CJD와 같은 다양한 신경 퇴행성 질환을 유발시키는 단백질 병원체이다. 정상 prion 단백질인 PrP^C가 병원성 PrP^Sc로 바뀌는 과정에 대해서는 많은 연구가 진행되었고, PrP^Sc로의 단백질 구조 변화가 다양한 환경적 요소에 의해서 영향 받는 것으로 추측된다. 바다조류로부터 분리된 MAAs는 다양한 스트레스 환경에서 조류를 보호해주는 것으로 알려져 있다. 이와 같은 사실에 기초하여 mycosporineglycine, porphyra-334와 shinorine 3종의 MAAs로 처리한 prion 감염 신경세포 주에서 prion 단백질 축적의 변화를 평가하였다. PK 저항성을 갖는 PrP^Sc를 western blot 방법으로 확인한 결과, MAA에 의해서 PrP^Sc 단백질의 증식을 관찰하였다.

• Journal of Microbiology and Biotechnology
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• 제24권2호
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• pp.245-253
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• 2014

A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDS-PAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the ${\alpha}$ and ${\beta}$ chains of fibrinogen followed by the ${\gamma}$ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at $45^{\circ}C$ and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

• Journal of Microbiology and Biotechnology
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• 제25권9호
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• pp.1493-1501
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• 2015

A Bacillus species, EMD4, with strong antibacterial activity was isolated from ganjang (soy sauce) and identified as B. subtilis. B. subtilis EMD4 strongly inhibited the growth of B. cereus ATCC14579 and B. thuringiensis ATCC33679. The antibacterial activity was stable at pH 3-9 but inactive at pH 10 and above. The activity was fully retained after 15 min at 80℃ but reduced by 50% after 15 min at 90℃. The activity was completely destroyed by proteinase K and protease treatment, indicating its proteinaceous nature. The bacteriocin (BacEMD4) was partially purified from culture supernatant by ammonium sulfate precipitation, and Q-Sepharose and Sephadex G-50 column chromatographies. The specific activity was increased from 769.2 AU/mg protein to 8,347.8 AU/mg protein and the final yield was 12.6%. The size of BacEMD4 was determined to be 3.5 kDa by Tricine SDS-PAGE. The N-terminal amino acid sequence was similar with that of Subtilosin A. Nucleotide sequencing of the cloned gene confirmed that BacEMD4 was Subtilosin A. BacEMD4 showed bactericidal activity against B. cereus ATCC14579.

• Asian-Australasian Journal of Animal Sciences
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• 제17권8호
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• pp.1062-1068
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• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

• Asian-Australasian Journal of Animal Sciences
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• 제21권2호
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• pp.270-276
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• 2008

The purpose of the present study was to investigate the effects of exogenous non-starch polysaccharides (NSP) enzymes on performance, intestinal content viscosity and digestive enzyme activities of growing pigs fed a rough rice-based diet. A total of 60 crossbred barrows with an initial body weight of 35.16 kg (SD = 0.82) were blocked by body weight and randomly assigned to two treatments with three replications. Each group was fed the diet based on rice with or without exogenous NSP enzymes (2 g/kg of diet). During the 70 days of the feeding trial, all pigs were given free access to feed and water. At the end of the feeding trial, six pigs from each treatment were randomly selected and slaughtered to collect intestinal digesta, intestinal mucosa, and pancreas. The addition of NSP enzymes improved average daily gain (p<0.05) and feed:gain (p<0.05), and decreased viscosity of digesta in the jejunum (p<0.001) and ileum (p<0.01) of pigs. The supplementation of NSP enzymes increased activities of protease (p<0.01), trypsin (p<0.01) and ${\alpha}$-amylase (p<0.05) in duodenal contents. However, digestive enzymes in the pancreas, jejunal and ileal mucosa were unaffected by the supplemental NSP enzymes (p>0.10). The results indicate that the addition of NSP enzymes to rough rice-based diets improved performance of pigs, reduced viscosity and increased digestive activity in the small intestine.

• 동아시아식생활학회지
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• 제12권4호
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• pp.326-333
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• 2002

본 연구는 능이버섯분말의 단백분해 활성을 육류연화에 활용할 수 있는 가능성을 검토한 것이다. 능이버섯을 동결건조와 열풍건조하여 분말을 만들었을 경우 동결건조분말의 단백분해효소활성이 높았으며 분말을 저장할 경우에도 -2$0^{\circ}C$에서 장시간 효소활성을 보존할 수 있었다. 능이버섯가루를 스테이크용 쇠고기에 처리할 때 4$^{\circ}C$보다는 상온에서 처리할 경우 경도가 더 빨리 감소하였으며 따라서 가열감량도 급속히 증가하였다. 기호도 측면에서 확립된 능이버섯가루의 처리농도는 0.1%, 처리시간은 4$^{\circ}C$에서 3시간이 최적으로 나타났다. 상업용연화제로 처리할 경우와 비교하였을 때 기호도면에서 능이버섯가루를 처리하는 것이 전반적으로 우수하였으며 능이버섯분말을 천연연육제로 개발 할 수 있는 가능성을 확인할 수 있었다.

• BMB Reports
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• 제31권5호
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• pp.459-467
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• 1998

A new set of functional group interaction energy descriptors relevant to the ACE (Angiotensin-Converting Enzyme) inhibitory peptide, QSAR (Quantitative Structure Activity Relationships), is presented. The functional group interaction energies approximate the charged interactions and distances between functional groups in molecules. The effective energies of the computationally derived geometries are useful parameters for deriving 3D-QSAR models, especially in the absence of experimentally known active site conformation. ACE is a regulatory zinc protease in the renin-angiotensin system. Therapeutic inhibition of this enzyme has proven to be a very effective treatment for the management of hypertension. The non bond interaction energy values among functional groups of six-feature of ACE inhibitory peptides were used as descriptor terms and analyzed for multivariate correlation with ACE inhibition activity. The functional group interaction energy descriptors used in the regression analysis were obtained by a series of inhibitor structures derived from molecular mechanics and semi-empirical calculations. The descriptors calculated using electrostatic and steric fields from the precisely defined functional group were sufficient to explain the biological activity of inhibitor. Application of the descriptors to the inhibition of ACE indicates that the derived QSAR has good predicting ability and provides insight into the mechanism of enzyme inhibition. The method, functional group interaction energy analysis, is expected to be applicable to predict enzyme inhibitory activity of the rationally designed inhibitors.

• Fisheries and Aquatic Sciences
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• 제20권3호
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• pp.3.1-3.8
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• 2017

Seahorse, a syngnathidae fish, is one of the important organisms used in Chinese traditional medicine. Hippocampus abdominalis, a seahorse species successfully cultured in Korea, was validated for use in food by the Ministry of Food and Drug Safety in February 2016; however. the validation was restricted to 50% of the entire composition. Therefore, to use H. abdominalis as a food ingredient, H. abdominalis has to be prepared as a mixture by adding other materials. In this study, the effect of H. abdominalis on muscles was investigated to scientifically verify its potential bioactivity. In addition, the anti-fatigue activity of a mixture comprising H. abdominalis and red ginseng (RG) was evaluated to commercially utilize H. abdominalis in food industry. H. abdominalis was hydrolyzed using Alcalase, a protease, and the effect of H. abdominalis hydrolysate (HH) on the muscles was assessed in C2C12 myoblasts by measuring cell proliferation and glycogen content. In addition, the mixtures comprising HH and RG were prepared at different percentages of RG to HH (20, 30, 40, 50, 60, 70, and 80% RG), and the anti-fatigue activity of these mixtures against oxidative stress was assessed in C2C12 myoblasts. In C2C12 myoblasts, $H_2O_2$-induced oxidative stress caused a decrease in viability and physical fatigue-related biomarkers such as glycogen and ATP contents. However, treatment with RG and HH mixtures increased cell viability and the content of fatigue-related biomarkers. In particular, the 80% RG mixture showed an optimum effect on cell viability and ATP synthesis activity. In this study, all results indicated that HH had anti-fatigue activity at concentrations approved for use in food by the law in Korea. Especially, an 80% RG to HH mixture can be used in food for ameliorating fatigue.