• Journal of Nutrition and Health
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• 제25권3호
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• pp.233-247
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• 1992

The changes in human milk composition from 2-5 days to 12 weeks postpartum were investigated. Milk from 62 mothers was anlyzed for total nitrogen(semimicro kjeldahl) lipid(utilizing a modified Folch) and lactose(enzymatic hydrolysis) Energy was calculated by frac-tional analysis. And the daily milk intakes and major nutrients and energy intakes of 18 exclusi-vely breast-fed infant were determined by the test-weighing procedure and the direct analysis of milk samples at 6 or 7 weeks postpartum. All samples were from well-defined subjects and uniform collection procedures were used. Total nitrogen content decreased significantly from 392 to 211 mg/dl lipid and lactose content increased from 1.94 to 3.06g/dl and 6.90 to 7.50g/dl respectively. And energy content increased 55.6 to 64.5 kcal/dl but was not statistically significant. The amount of milk ingested ranged from 432 to 1266 ml/day and the mean intake was 764 ml/day. Daily mean intakes for protein and energy were 10.0g and 450kcal in 6 or 7 weeks postpartum respectively.

• Journal of Microbiology and Biotechnology
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• 제29권5호
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• pp.749-757
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• 2019

Nitrilase is a valuable hydrolase that catalyzes nitriles into carboxylic acid and ammonia. Its applications, however, are severely restricted by the harsh conditions of industrial reaction processes. To solve this problem, a nitrilase from Acidovorax facilis 72W was inserted into an Escherichia coli-Bacillus subtilis shuttle vector for spore surface display. Western blot, enzyme activity measurements and flow cytometric analysis results all indicated a successful spore surface display of the CotB-nit fusion protein. In addition, the optimal catalytic pH value and temperature of the displayed nitrilase were determined to be 7.0 and $50^{\circ}C$, respectively. Moreover, results of reusability tests revealed that 64% of the initial activity of the displayed nitrilase was still retained at the $10^{th}$ cycle. Furthermore, hydrolysis efficiency of upscale production of cyanocarboxylic acid was significantly higher in the displayed nitrilase-treated group than in the free group expressed by E. coli (pET-28a-nit). Generally, the display of A. facilis 72W nitrilase on the spore surface of Bacillus subtilis may be a useful method for immobilization of enzyme and consequent biocatalytic stabilization.

• Asian-Australasian Journal of Animal Sciences
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• 제32권5호
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• pp.629-636
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• 2019

Objective: The study was designed to determine the cytotoxic effect of gallic acid (GA), obtained by the hydrolysis of tannins, on mice TM4 Sertoli cells apoptosis. Methods: In the present study, non-tumorigenic mice TM4 Sertoli cells were treated with different concentrations of GA for 24 h. After treatment, cell viability was evaluated using WST-1, mitochondrial dysfunction, cells apoptosis and necrosis was detected using JC-1, Hoechst 33342 and propidium iodide staining. The expression levels of Cyclin B1, proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (BAX), and Caspase-3 were also detected by quantitative real-time polymerase chain reaction and Western-blotting. Results: The results showed that 20 to $400{\mu}M$ GA inhibited viability of TM4 Sertoli cells in a dose-dependent manner. Treatment with $400{\mu}M$ GA significantly inhibited PCNA and Cyclin B1 expression, however up-regulated BAX and Caspase-3 expression, caused mitochondrial membrane depolarization, activated Caspase-3, and induced DNA damage, thus, markedly increased the numbers of dead cells. Conclusion: Our findings showed that GA could disrupt mitochondrial function and caused TM4 cells to undergo apoptosis and necrosis.

• 한국환경농학회지
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• 제40권1호
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• pp.20-32
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• 2021

BACKGROUND: Bacterial shot hole of stone fruits is a seriuos plant disease caused by Xanthomonas arboricola pv. pruni (Xap). Techniques to control the disease are required. In this study, microorganisms with antibacterial activity were isolated to develop as a microbial agent against the bacterial shot hole. METHODS AND RESULTS: An isolate with the strongest activity among the isolates was identified as Streptomyces avidinii based on 16S rRNA gene sequence analysis and designated Streptomyces sp. J46. J46 showed suppression of bacterial leaf spot with a control value of 90% at 10 times-diluted cell free supernatant. To investigate antibacterial metabolites produced by J46, the supernatant of J46 was extracted with organic solvents, and the extracts were subjected to chromatography works. Antibacterial metabolites were not extractable with organic solvents. Both reverse and normal phase techniques were not successful because the metabolites were extremely water soluble. The antibacterial metabolites were not volatiles but protein compounds based on hydrolysis enzyme treatment. CONCLUSION: Our study suggests that Streptomyces sp. J46 may be a potential as an microbial agent against bacterial shot hole. Further study to identify the metabolites is required in more detail.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.71-80
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• 2022

Lipases (triacylglycerol acylhydrolases [EC 3.1.1.3]) are water-soluble enzymes. They catalyze the hydrolysis of fats and oils. A cold-active lipase from marine Bacillus cereus HSS, isolated from the Mediterranean Sea, Alexandria, Egypt, was purified and characterized. The total purification depending on lipase activity was 438.9 fold purification recording 632 U/mg protein. The molecular weight of the purified lipase was estimated to be 65 kDa using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The optimum substrate concentration, enzyme concentration, pH, and temperature were 1.5 mM, 100 µl, pH 6 and 10℃, respectively. The lipase was tolerant to NaCl concentrations ranging from 1.5 to 4.5%. The lipase was affected by the tested metal ions, and its activity was inhibited by 16% in the presence of 0.05 M SDS. The application of the cold-active lipase for the removal of an oil stain from a white cotton cloth showed that it is a promising biological agent for the treatment of oily wastes and other related applications. To the best of our knowledge, this is the first report of the purification and characterization of a lipase from marine B. cereus HSS isolated from the Mediterranean Sea.

• Journal of Applied Biological Chemistry
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• 제65권2호
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• pp.93-99
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• 2022

Lysosome organelle extract (LOE) was derived from egg whites. The lysosome is an intracellular organelle that contains several hydrolysis enzymes. Previous studies have reported that LOE performs important functions, such as melanin de-colorization and anti-melanin production in B16F10 melanoma cells. However, its principal molecular and cellular mechanisms have not been elucidated till date. In non-cytotoxic conditions, LOE significantly inhibited α-MSH induced melanin synthesis of murine B16F10 cells. The anti-melanogenic activity of LOE was mediated by suppressing the mRNA expression of tyrosinase enzyme, tyrosinase related protein-1/2 (TRP-1/2), and microphthalmia-associated transcription factor (MITF) genes. By performing western blot analysis, we found that LOE significantly attenuated melanogenesis. In this case, LOE helped in increasing extracellular receptor kinase (ERK) phosphorylation in α-MSH induced B16F10 cells. Furthermore, MITF is found to be a key regulatory transcription factor in melanin synthesis; it was down-regulated by LOE through ERK phosphorylation. In this experiment, PD98059 (MEK inhibitor) was used to check whether LOE directly regulated the activity of ERK. Although LOE exerted inhibitory effect on melanin synthesis, we could not observe this effect in PD98059-treated α-MSH induced B16F10. These results strongly indicate that LOE is related to ERK activation and MITF degradation in anti-skin pigmentation. Hence, LOE should be utilized as a whitening agent of skin in the near future.

• 한국식생활문화학회지
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• 제23권1호
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• pp.121-129
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• 2008

To produce buckwheat sprouts, buckwheats were germinated at $23{\pm}2^{\circ}C$ up to 8 days in total darkness. In proximate analysis, moisture content increased from 13.87% of buckwheat groats to 93.75% of buckwheat sprouts on the 8th day of germination. On dry weight basis, lipid and ash contents increased like as protein content increased from 13.45% to 21.82% while carbohydrate content decreased due to enzyme hydrolysis. Amino acids were rich in glutamic acid, aspartic acids, and lysine, and also the ratio of the essential amino acids to total amino acids increased from 26.84% to 36.84%. Vitamin C did not exist in buckwheat groats but its content has continued to increase as far as 99.56 mg/100 g buckwheat sprouts. Rutin continued to increase by HPLC analysis from 4.71 mg/100 g buckwheat groats to 1,524 mg/100 g buckwheat sprouts. In analysis of organic acids, oxalic, maleic, and citric acids were commonly found in buckwheat sprouts. Fagopyrin was found almost none in buckwheat sprouts. In conclusion, the conversion of buckwheat seeds into sprouts through germination in darkness results in physically different final product with nutritional changes such as higher content of rutin, generation of vitamin C, abundance of the essential amino acids, and the existence of fagopyrin on very little.

• Journal of Animal Science and Technology
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• 제65권4호
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• pp.890-893
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• 2023

Lactobacillus johnsonii 7409N31 was isolated from the feces of a healthy 11-day-old Hanwoo calf from a farm in Geochang-gun, Gyeongsangnam-do, Korea. The genome of the strain was completely sequenced using the PacBio RSII sequencing system, and it was confirmed that it was composed of one circular chromosome. The size of the entire genome was 2,198,442 bp, and it had 35.01 mol% guanine + cytosine (G + C) content and 2,222 protein-coding sequences, 24 rRNA, 3 ncRNA, and 112 tRNA genes. Strain 7409N31 possessed genes encoding enzymes involved in the hydrolysis of both fibrous and non-fibrous carbohydrates. These data provide a comprehensive theoretical understanding for developing industrial probiotic feed additives that improve nutrient digestibility.

• 생명과학회지
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• 제16권3호
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• pp.409-414
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• 2006

소포체는 세포막의 합성뿐만 아니라 세포막에 존재하거나 세포외로 분비되어져야 할 단백질을 합성하는 세포내 소기관이다. 소포체에서 단백질이 합성되어질 경우 이황화결합이 형성되고 glycosylation 등의 수식이 일어나며, 이와 동시에 folding과 assembly과정을 거쳐 삼차원적 구조로 성숙이 되는데 이 과정은 folding enzyme과 molecular chaperone의 도움을 받아 이루어진다. 소포체 내에 존재하는 molecular chaperone 중 가장 잘 알려진 것으로 BiP이 있다. BiP의 기능은 N-terminus의 ATPase domain에 의해 조절되고 ATPase domain은 이것과 선택적으로 결합하는 조절인자에 의해 ATPase의 활성이 영향을 받는다. BiP의 핵산치환조절인자로서 발견된 BAP은 ATPase domain에 결합된 ADP를 ATP로 치환하는 것으로 기능이 알려져 있다. 이 BAP의 핵산치환기능이 BiP의 샤페론 작용에 어떤 영향을 미치는지를 in vitro에서 항체 heavy chain을 이용하여 알아보았다. BAP은 ATP보다 ADP가 결합되어 있는 BiP과 더 잘 결합을 하며, in vitro에서 BiP과 결합하고 있는 unfolded 단백질을 BAP은 BiP으로부터 해리하였다. 또한 소포체내에 존재하는 Hsp70 homologue chaperone인 BiP과 Grp170에 대한 BAP의 결합특이성을 anti-Grp170과 anti-BAP 항체로 co-immunoprecipitation을 하여 확인한 결과 BAP은 Grp170과 결합을 하지 않았다. 따라서 BAP은 ER내에 존재하는 동일한 family group에 속하는 Grp170과 BiP에 대하여 BiP에만 특이성을 갖는 것으로 나타났다.

• 한국응용과학기술학회지
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• 제34권3호
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• pp.673-682
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• 2017

식물 및 동물성 단백질 유래 펩타이드 형태의 단백질 가수 분해물들은 항산화, 고혈압 완화, 면역조절, 진통완화 및 항균작용 등 생리활성이 있는 것으로 알려져 왔다. 본 연구는 연산오계란 단백질 가수 분해물을 Ultrafiltration를 이용하여 HDS(분획되지 않은 가수 분해물), 1 kDa, 5 kDa, 10 kDa, 50 kDa로 분획된 기능성 펩타이드의 DPPH radical scavenging activity, superoxide anion radical scavenging activity, hydroxyl radical scavenging activity 및 $Fe^{2+}$ chelation ability을 평가하였다. 그 결과 DPPH radical scavenging activity 최대값은 1 kDa(70.83 %), hydroxyl radical scavenging activity 최대값은 5 kDa (47.01 %), superoxide anion radical scavenging activity 최대값은 5 kDa(40.57 %), $Fe^{2+}$ chelation ability 최대값은 5 kDa(29.87 %)로 나타났다. Ultrafiltration를 이용하여 fractionation된 단백질 가수 분해물의 항산화 저해 능력 $IC_{50}$ 평가하였다. 그 결과 HDS의 최대값은 superoxide anion radical scavenging activity($IC_{50}$, 5.42 mg/ml)이고, 1 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 1.67 mg/ml)이고, 5 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 2.09 mg/ml)이고, 10 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 2.61 mg/ml)이고, 50 kDa의 최대값은 $Fe^{2+}$ chelation ability($IC_{50}$, 4.53 mg/ml)이다. 그러므로 본 연구 결과를 바탕으로 5 kDa를 이용하여 오계란 단백질에서 분획한 펩타이드는 항산화 기능성 식품소재로서 활용할 가치가 높을 것으로 기대한다.