• 한국축산식품학회지
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• 제38권6호
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• pp.1294-1304
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• 2018

The aims of this research were to examine the effect of heating temperature (65, 75, and $85^{\circ}C$) and $CaCl_2$ concentration level (3, 4, and 5 mM) on the physicochemical properties of ${\beta}$-lactoglobulin (${\beta}$-lg) nanoemulsions (NEs) and to study how the droplet size of NEs affects the bioaccessibility (BA) of coenzyme $Q_{10}$ ($CoQ_{10}$). The droplet size of NEs and BA of $CoQ_{10}$ was assessed by particle size analyzer and UV-Vis spectrophotometer, respectively. An increase in heating temperature and $CaCl_2$ concentration level resulted in a significant (p<0.05) increase in the droplet size of NEs while there were no significant differences in polydispersity index and zeta-potential of NEs. When NEs containing $CoQ_{10}$ were incubated in simulated small intestinal phases, an increase in the droplet size and polydispersity index of NEs was observed. This indicated that NEs were not stable in small intestine and digestion of NEs occurred. As heating temperature and $CaCl_2$ concentration level were decreased, a significant (p<0.05) increase in BA of $CoQ_{10}$ was observed. There was a significant (p<0.05) increase in BA of $CoQ_{10}$ with a decrease in the droplet size of NEs. In conclusion, heating temperature and $CaCl_2$ concentration level were key-parameters affecting the initial droplet size of NEs and BA of $CoQ_{10}$ was negatively correlated with initial droplet size of NEs.

• BMB Reports
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• 제29권3호
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.40-49
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• 2001

The present work presents a novel approach for the dynamic quantification of respiration rates on a small scale by using lysine-producing Corynebacterium glutamicum ATCC 21253. Cells sampeld from batch cultures at different times were incubated ina 12-ml scale bioreactor equipped with a membrane mass spectrometer. Under dynamic conditions, gas exchange across the gas-liquid phase, specific respiration rates, and RQ values were precisely measured. For this purpose, suitable mass balances were formulated. The transport coefficients for $O_2$ and $CO_2$, crucial for calculating the respiration activity, were determined as $k_La_{O2}=9.18h^{-1}$ and $k_La_{CO2}=5.10h^{-1}$ at 400 rpm. The application of the proposed method to batch cultures of C. glutamicum ATCC 21253 revealed the maximum specific respiration rates of $q_{O2}=8.4\;mmol\;g^{-1}h^{-1}\;and\;q_{CO2}=8.7\;mmol\;g^{-1}h^{-1}$ in the middle of the exponential growth phase after 5 h of cultivation. When the cells changed from growth to lysine production due to the depletion of the essential amino acids theonine, methionine, and leucine, $q_{O2}\;and\;q_{CO2}$ decreased significantly and RQ increased. The respiration data exhibited an excellent agreement with previous cultivations of the strain [13]. This confirms the potential of the developed approach to realistically reflect the metabolic activities of cells at their point of sampling. The short-term influence of added threonine, methionine, and leucine was highest during the shift from growth to lysine production, where $q_{O2}\;and\;q_{CO2}$ increased 50% within one minute after the pulse addition of these compounds. Non-growing, yet lysine-producing cells taken from the end of the batch cultivation revealed no metabolic stimulation with the addition of threonine, methionine, and leucine.

• 한국수정란이식학회지
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• 제11권1호
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• pp.7-14
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• 1996

The object of this study was to evaluate the effect of uterine epithelial cells on development of Korean native cattle(KNC) oocytes fertilized in vitro. Qocytes were collected from ovaries of slaughtered Korean Native Cows and matured in TCM199 with granulosa cells supplemented with 10% FBS, 5$\mu$g/ml FSH, 10 JU/ml hCG, and 1$\mu$g/ml estradiol-17$\beta$ for 24 hrs. For co-culture of in vitro development of fertilized ova, oviductal epithelial cells (l$\times$l0˚cells /ml) obtained from slaughtered cow and uterine epithelial cells(1$\times$10˚cells /ml) flushed from the superovulated holstein on Day 7 were incubated in 39$^{\circ}C$, 5% $CO_2$, 95% air. Frozen-thawed KNC sperm was capacitated with BO(Brackett & Oliphant, 1975) medium supplemented with 10mM, 5mM-caffein. Matured oocytes were inseminated for 20 hrs. And then fertilized oocytes were washed with culture medium and transferred to oviductal epithelial cells for in vitro development and three days later a portion of embryos were transferred to uterine epithelial cells. Stastical methods of developmental rates on KNC-IVF oocytes was ANOVA-test. Developmental rates of KNC-IVF oocytes was significant higher(P<0.01) when co-cul-tured with uterine epithelial cells(25.2%) than oviductal epithelial cells. Blatocyst cul-tured for 7 to 9 days were frozen by automatic freezer with 1.4M glycerol-PBS. Survival rates of blastocyst was 40.0%. Fourteen frozen-thawed blastocysts were transferred to five holstein heifers on day 7 after natural estrus. Three recipients were observed twin and one recipient was single by ultra-sound systems on days 45 after embryo transfer.

• Journal of Animal Science and Technology
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• 제65권2호
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• pp.387-400
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• 2023

Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39℃ for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

• 한국환경농학회지
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• 제17권1호
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• pp.5-10
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• 1998

물리화학적 특성이 상이한 4종의 토양을 이용하여 제초제 imazapyr의 미생물에 의한 분해, 감광제에 의한 광분해 촉진 및 bioceramic 첨가에 의한 분해 촉진 시험을 수행하여 다음과 같은 결과를 얻었다. 토양 A와 active sludge로부터 분리한 7종의 미생물을 접종한 순수배양에서 뚜렷한 대사산물을 얻지 못했다. 또한 6종의 기지 미생물을 이용한 14일간의 배양실험에서 역시 대사산물을 얻지 못했다. 이 결과는 수중에서 imazapyr가 미생물에 의해서는 거의 분해가 되지 않음을 시사해 준다. Imazapyr를 처리하여 배양한 토양중 그 분해산물로는 imidazolinone ring의 개열에 의해 형성된 m/z 279의 2-[(1-carbamoyl-1,2-dimethylpropyl)carbamoyl]nicotinic acid를 얻었다. 또한 자연광하에서 행한 광분해 시험에서 감광제의 종류에 따라 분해율에 많은 차이를 보였다. 무처리의 경우에는 14.6%의 분해율을 보인 반면 PS-1의 100ppm과 200ppm에서 각각 66.0과 76.5%로 농도가 높을수록 높은 분해율을 보였고, PS-2와 PS-3에서 각각 26.7과 90.0%의 분해율을 보였다. Aromatic ketone계 감광제인 PS-2는 무처리와 큰 차이를 보이지 않았다. PS-1을 첨가한 광분해 시료에서 m/z 149의 광분해 산물을 검출하였으며, 그 생성경로는 imidazolinone환이 개열된 후 가수분해되어 2-carbamoyl-nicotinic acid ${\rightarrow}$ 2,3-pyridinedicarboxylic acid ${\rightarrow}$ 2,3-pyridine-dicarboxylic anhydride(m/z 149)로 추정되었다. 토양 C와 D에 $[^{14}C]$imazapyr를 처리하고 bioceramic을 첨가하였을 때 발생된 $^{14}CO_2$의 방사능은 각각 총 처리방사능의 2.03%와 1.12%인 반면 bioceramic을 처리하지 않은 구에서는 각각 1.88%와 0.82%로써 유의성 있는 차이는 보이지 않았으나 5주 이후에는 $^{14}CO_2$의 양이 점점 증가했다.

• Nutrition Research and Practice
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• 제3권3호
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• pp.192-199
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• 2009

Cadmium intoxication has been associated with the dysregulation of iron homeostasis. In the present study, we investigated the effect of cadmium on the expression of ferroportin 1 (FPN1), an important iron transporter protein that is involved in iron release from macrophages. When we incubated cadmium with J774 mouse macrophage cells, FPN1 mRNA levels were significantly increased in a dose- and time-dependent manner. Furthermore, the cadmium-induced FPN1 mRNA expression was associated with increased levels of FPN1 protein. On the other hand, cadmium-mediated FPN1 mRNA induction in J774 cells was completely blocked when cells were co-treated with a transcription inhibitor, acitomycin D. Also, cadmium directly stimulated the activity of the FPN1-promoter driven luciferase reporter, suggesting that the cadmium up-regulates FPN1 gene expression in a transcription-dependent manner. Finally, cadmium exposure to J774 macrophages increased intracellular reactive oxygen species (ROS) levels by ${\sim}2$-fold, compared to untreated controls. When J774 cells were co-treated with antioxidant N-acetylcystein, the cadmium-induced FPN1 mRNA induction was significantly attenuated. In summary, the results of this study clearly demonstrated that cadmium increased FPN1 expression in macrophages through a mechanism that involves ROS production, and suggests another important interaction between iron and cadmium metabolism.

• Journal of Microbiology and Biotechnology
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• 제2권3호
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• pp.209-214
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• 1992

Serratia marcescens co-cultured with various phytopathogenic fungi, including Rhizopus stolonifer, Helminthosporium allii, Pyricularia oryzae, Fusarium oxysporium and Collectothricom cassiicola, in an LB- agar medium containing 1.5% swollen chitin, significantly inhibitied fungal growth. Fungal hyphae grew rapidly outward from the culture dish center, but the hyphal extensions of the pathogenic fungi were significantly inhibited in a perimetric contact area with S. marcescens. This was especially evident in pathogenic fungi which have a high chitin content in their cell walls. The extracellular chitinase activities of S. marcescens were increased seven fold by the addition of 1.5% swollen chitin to the LB-broth, compared to chitinase activities in a culture medium without chitin. The type of induction was dependent on the various forms of chitin used. When the culture supernatant of S. marcescens or the chitinases of Streptomyces griceus purchased from Sigma Chemical Co., were incubated with the mycelium of F. oxysporium, the mycelium gradually burst as cultivation time progressed and completely lysed after incubation for 2 days. On the other hand, E. coli extract did not hydrolyze the F. oxysporium mycelium at all. These data showed that the chitinolytic activities of S. marcescens play important roles in the biochemical control of phytopathogenic fungi.

• Applied Biological Chemistry
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• 제25권3호
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• pp.126-134
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• 1982

$^{14}C-2$, 6-Diethylaniline을 토양중에서 호기적으로 21 주간 배양시 발생된 $^{14}CO_2$는 살균하지 않은 A토양(clay loam)과 B토양(coarse sandy loam)에서 각각 6.5%와 10.1%이었다. Methanol로 토양을 추출시 A토양에서는 3.1%, B토양에서는 13.5%의 토양방사능이 추출되었다. 토양중에서의 분해산물은 2, 6-diethylacetanilide 이었으며 Chaetomium globosum은 순수배양시 분해산물로 2, 6-diethyl-p-benzoquinone을 생성하였고 이의 생성경로는 2, 6-DEA의 p-hydroxylation, quinoneimine의 형성 그리고 암모니아 발생을 수반하는 가수분해 등으로 제안하였다.

• BMB Reports
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• 제35권2호
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• pp.228-235
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• 2002

Methionine aminopeptidase (MetAP) catalyzes the removal of an amino-terminal methionine from a newly synthesized polypeptide. The enzyme was purified to homogeneity from Bacillus stearothermophilus (KCTC 1752) by a procedure that involves heat precipitation and four sequential chromatographs (including DEAE-Sepharose ion exchange, hydroxylapatite, Ultrogel AcA 54 gel filtration, and Reactive red 120 dye affinity chromatography). The apparent molecular masses of the enzyme were 81,300 Da and 41,000 Da, as determined by gel filtration chromatography and sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. This indicates that the enzyme is comprised of two identical subunits. The MetAP specifically hydrolyzed the N-terminal residue of Met-Ala-Ser that was used as a substrate, and exhibited a strong preference for Met-Ala-Ser over Leu-Gly-Gly, Leu-Ser-Phe, and Leu-Leu-Tyr. The enzyme has an optimal pH at 8.0, an optimal temperature at $80^{\circ}C$, and pI at 4.1. The enzyme was heat-stable, as its activity remained unaltered when incubated at $80^{\circ}C$ for 45 min. The Km and Vmax values of the enzyme were 3.0mM and 1.7 mmol/min/mg, respectively. The B. stearothernmophilus MetAP was completely inactivated by EDTA and required $Co^{2+}$ ion(s) for activation, suggesting the metal dependence of this enzyme.