• Asian-Australasian Journal of Animal Sciences
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• 제15권2호
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• pp.222-237
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• 2002

Effects of high and low levels of feeding with or without protected protein on the performance of lactating goats were studied. Twenty four German Fawn goats either from 1st ($43.37{\pm}3.937$ kg and 2 year old) or 3rd $62.64{\pm}6.783$ kg and 4-5 year old) parity were used for the trial. Feeding levels were 7.2 (I) and 5.2 (II) MJ ME/litre of milk of 3.5% fat in addition to that of the maintenance allowance. At each feeding level, diet had either unprotected (U) or formaldehyde protected (P) soya-meal. Thus, four diets were IU, IP, IIU and IIP, having six animals in each. The diets were composed of hay and pellet (10:4:1 of beet pulp : barley : soya-meal). Effect of feeding level, protein protection, parity, health status and kid number on intake, milk yield, milk composition, growth rate of goats were recorded across the 21 weeks of study. High feeding level resulted increase (p<0.001) in estimated metabolizable energy (ME) and metabolizable protein (MP) availability. Dietary inclusion of protected soya-meal increased (p<0.001) the estimated MP but not the ME availability. Animals in 1st parity ate more (p<0.001) DM (111 vs. 102 g/kg $W^{0.75}$/d) than those in 3rd parity. Animals with twin kids (110 g/kg $W^{0.75}$/d) had higher (p<0.001) DM intake than those with single kid (102 g/kg $W^{0.75}$/d). Fat (4%) corrected milk (FCM) yield was not effected by high (1,924 g/d) or low (1,927 g/d) feeding level but increased (p<0.001) with protected (2,166 g/d) compared with unprotected (1,703 g/d) soya-meal. FCM yield for four dietary combinations were 1,806, 2,078, 1,600 and 2,254 g/d for diets IU, IP, IIU and IIP, respectively. For unit increase (g) in estimated MP availability relative to ME (MJ) intake, FCM yield increased ($1,418{\pm}275.6$) g daily ($r^2$=0.58; p<0.001). Milk fat (3.14 vs. 3.54%; p<0.001) and protein (2.94 vs. 3.04% p<0.05) contents were lower at high than the low feeding level. Protected protein increased (p<0.001) the fat, lactose and net energy (NE) content of milk. Milk urea concentration of 175, 183, 192 and 204 mg/l for diets IU, IP, IIU and IIP, respectively indicated lower RDP content of these diets. The RDP contents were 6.97, 6.70, 7.30 and 6.83 g/MJ of ME for diets IU, IP, IIU and IIP, respectively. Live weight change over the experimental period were 41, 6, 17 and 19 g/d. Absence of any positive response of high feeding was probably due to inefficient rumen fermentation resulting from inadequate RDP supply. Protected protein improved production performance apparently by increasing MP:ME ratio in the absorbed nutrient.

• 한국축산식품학회지
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• 제38권5호
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• pp.1101-1108
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• 2018

The aim of this study was to investigate the effects of dry-aging (DA) and the cooking process on the myofibril protein functionalities and in vitro digestibility of proteins in beef loin. Six sirloins from beef were dry-aged for 28 d, and the control group (n=6) was analyzed 2 d postmortem for this study. Dimensional changes (reduction of thickness and surface shrinkage) after cooking were significantly greater in the control group than the DA group, whereas the shear force of the DA group was significantly lower than that of the control. Effect of cooking on aggregation, hydrophobicity, and in vitro digestibility were significantly higher in the DA group than in the control. After cooking, the protein in DA sirloins was more oxidized than in the control samples. According to the sodium dodecyl sulfate-polyacrylamide gel electrophoresis result, the low molecular weight bands (below 17 kDa) increased in the DA group, finding that the protein characteristics of dry-aged beef was affected by cooking.

• Applied Biological Chemistry
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• 제43권3호
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• pp.174-178
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• 2000

인체 세포의 세포주기를 조절하는 Cyclin-dependent Kinase(CDK) 중 CDK4/Cyclin D1 효소 저해 물질을 찾기 위하여 국내에 자생하는 37과 87종의 식물 추출물을 탐색한 결과, 50%$(5\;{\mu}g/ml)$ 이상의 CDK4/Cyclin D1 호소 저해활성을 보인 것은 단삼(Salvia miltiorrhiza), 노박덩굴(Celastrus orbiculatus), 삼백초(Saurus chinensis), 목단(Paeonia suffruticosa) 및 오이풀(Sanguisorba officinalis)이었다. 이들 중에서 목단과 오이풀 추출물이 가장 높은 56%의 저해율을 각각 나타내었다. 또한 추출물에서 활성물질을 추출 정제하여 2차원 NMR 등을 포함한 각종 기기분석을 통하여 cryptotanshinone으로 동정하였다.

• Journal of Applied Biological Chemistry
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• 제43권3호
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• pp.125-130
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• 2000

An antiviral protein (CAP30) with ribosome-inactivating activity was purified from the leaves of Chenopodium album L. through ammonium sulfate precipitation and column chromatography using S-Sepharose, Blue-Sepharose, FPLC Suprose12 HR, and FPLC Mono-S. The molecular wight of CAP30 was estimated to be 30kD. CAP30 was thermostable, maintaing its activity even after incubation at $70^{\circ}C$ for 30 min, and was stable in the pH range of 6 to 9. In a cell-free in vitro translation system using rabbit reticulocyte lysate, protein synthesis was inhibited by the addition of CAP30 with an $IC_{50}$ of 2.26pM. The comparison of N-terminal amino acid sequences of this protein with known ribosome-inactivating proteins (RIPs) revealed that it had some sequence homology with PAP-S and PAP-R from pokeweed (Phytolacca americana)and dodecandrin from P. dodecandra, but had no sequence homology with RIPs from other plants belonging to different orders. The mosaic symptoms on tobacco leaves caused by cucumber mosaic virus infection was completely inhibited by 100 ng/ml of the pure CAP30 protein.

• 한국식품과학회지
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• 제47권6호
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• pp.772-778
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• 2015

최근 아임계수 가수분해는 전통적인 단백질 가수분해법의 대체방법으로서 관심을 받고 있으며, 고단백질원으로부터 아미노산을 회수하는데 효과적인 공정이 될 수 있을 것으로 기대되고 있다. 본 연구에서는 대표적인 식물성 단백질원인 분리대두단백질을 선택하여 대두단백질의 아임계수 가수분해에서 가장 중요한 인자인 대두단백질의 초기농도, 반응온도, 반응시간의 영향을 연구하여 대두단백질로부터 아미노산을 생산할 수 있는 조건을 최적화하고자 하였다. 대두단백질 수열분해액의 실온에서 pH는 $200^{\circ}C$에서 20분간 처리했을 때 pH는 7.3으로 중성이었으나 온도가 증가할수록 pH가 증가하여 $270^{\circ}C$에서 10.3으로 알카리성을 나타내었다. 반응온도 $220^{\circ}C$까지는 수열분해액은 분산상태를 이루었으나 $230^{\circ}C$ 이상에서는 분산상태가 파괴되고 단백질이 분리되어 하부에 침강층을 이루었으며 $240^{\circ}C$ 이상에서는 이 단백질층이 현저히 감소하였다. 반응온도는 $250^{\circ}C$, 반응시간 20분으로 고정한 조건에서 대두현탁액의 초기농도가 수열분해에 미치는 영향을 살펴본 결과 초기농도 10% (w/v)일 때 가수분해도 및 아미노산 수율이 각각 16.2% 및 22.3%로 가장 우수하였다. 또한 반응온도 $200-220^{\circ}C$ 범위에서는 가수분해도와 아미노산 수율은 서서히 증가하였으나 $230-250^{\circ}C$ 영역에서는 급격히 증가하였으며 $250^{\circ}C$ 이상에서는 다시 완만히 증가하는 경향을 보여 $250^{\circ}C$ 이상에서 아미노산 분해속도는 단백질 분해속도를 초과하였다. 표면반응분석법으로 예측한 결과 $268^{\circ}C$, 처리시간 35분에서 최적 아미노산 수율 43.5%를 얻을 수 있었다.

• Reproductive and Developmental Biology
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• 제29권3호
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• pp.187-191
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• 2005

본 연구는 한우 난포란이 체외성숙된 환경의 변화를 단백질 측면으로부터 검토하기 위하여 체외성숙 배지와 세포질내 단백질 변화와 종류를 검토하였다. 그 결과 배지 내의 단백질 발현량은 배양 4.5시간째까지 감소하였고, 배양 13.5시간째까지는 변화가 없었다. 그러나 배양 $13.5\~18$시간 사이에 증가한 후 배양 18시간 이후에 다시 감소하는 경향이었다. 세포질내의 단백질 발현량은 배양 4.5시간째까지 증가하였고 배양 9시간째까지 급격히 감소하였다. 배양 9시간째부터 18시간째 까지는 단백질 발현량이 유사한 경향이었으나 배양 18시간째부터 24시간째까지 다시 증가하였다. 한편 체외성숙한 배지와 세포질을 2차원 전기영동하여 각각 298개 및 35개의 단백질 spot을 확인하였고, 그 중 배지에서는 28개, 세포질에서는 5개의 spot이 유의적인 변화를 확인하였다. 이들 spot 대한MALDI-TOP분석으로 배지와 세포질에서 각각 8개 및 1개의 단백질을 동정하였다. 그 종류는 aldose reductase, alpha enolase, apolipoprotein A-1 precursor, 43M1a collectin precursor, heat shock 27kDa protein, plasminogen activator inhibitor-1 precursor, thrombospondin 1 transitional endoplasmic reticulum ATPase 및 $\beta$-tubulin이었다.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1499-1512
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• 2006

Based on the first 2D reference map of the fission yeast Schizosaccharomyces pombe protein reported previously, we expanded and updated the map using narrower pI ranges. In this paper, 240 protein spots were identified on our reference map. In the pI 4-7 range, 144 spots corresponding to 86 different proteins were identified. In the pI 6-9 range, 43 spots corresponding to 35 different proteins were identified. Fifty-three new spots corresponding to 39 different proteins were further identified in the pI 5-6 range.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• 제3권3호
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• pp.177-188
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• 1999

The effect of ABA on the chloroplast disassembly of Zea mays was investigated by measuring the changes in the relative distribution of chlorophyll(Chl) between the Chl-protein complexes in ABA treated and untreated sensecting leaves. The reaction center(RC)-light harvesting complex(LHC) regions were rapidly disassembled in the late stage of dark-induced senescence. Plus, during dark-induced senescence, the disassembly of a reaction center of P700 apoproteins containing mainly Chl a was faster than that of a reaction center of LHCI apoproteins containing both Chl a and Chl b. The increase in the relative distribution of Chl-protein complexes in the RC-Core2 in the late stage of senescence was due to the accumulation of core complexes such as CP47/43 and reaction centers including D1/D2 apoproteins disassembled from the RC-Corel containing the dimer of D1/D2 apoproteins. The LHCII region was more stable than the other Chl-protein complexes throughout leaf senscence. Accordingly, it is suggested that the preferential breakdown of Chl a gives rise to the disassembly of Chl a-binding proteins, particularly reaction centers and core complexes during dark-induced senescence, plus the primary target of the photosynthetic apparatus in sensecing leaves would seem to be Chl a along with the proteins associated with Chl a. The application of ABA promoted the disassembly of the P700 apoproteins in the PSI reaction center and the dimer of D1/D2 apoproteins, and the conversion of the trimeric LHCII apoprotein to the monometirc LHCII apoprotein during the middle stage of leaf senescence, thereby suggesting that ABA accelerates the disassembly of both Chl a-binding and Chl a+b-binding proteins, particularly Chl a-binding proteins during the middle stage of leaf senescence.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권3호
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• pp.167-172
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• 2001

Staphylokinase (SAK) was produced in B. subtilis using two different promoter systems, i.e. the P43 and sacB promoters. To maximize SAK expression in B. subtilis, fermentation control strategies for each promoter were examined. SAK, under P43, a vegetative promoter transcribed mainly by $\sigma$(sup)B containing RNA polymerase, was overexpressed at low dissolved oxygen (D.O.) levels, suggesting that the sigB operon is somewhat affected by the energy charge of the cells. The expression of SAK at the 10% D.O. level was three times higher than that at the 50% D.O. level. In the case of sacB, a sucrose-inducible promoter, sucrose feeding was used to control the induction period and induction strength. Since sucrose is hydrolyzed by two sucrose hydrolyzing enzymes in the cell and culture broth, the control strategy was based on replenishing the loss of sucrose in the culture. With continuous feeding of sucrose, WB700 (pSAKBQ), which contains the SAK gene under sacB promoter, yielded ca. 35% more SAK than the batch culture. These results present efficient promoter-dependent control strategies in B. subtilis host system for foreign protein expression.

• Molecules and Cells
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• 제43권4호
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• pp.373-383
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• 2020

Our previous study revealed a novel role of Fas-associated death domain-containing protein (FADD) in islet development and insulin secretion. Insulin-degrading enzyme (IDE) is a zinc metalloprotease that selectively degrades biologically important substrates associated with type 2 diabetes (T2DM). The current study was designed to investigate the effect of FADD phosphorylation on IDE. We found that the mRNA and protein levels of IDE were significantly downregulated in FADD-D mouse livers compared with control mice. Quantitative real-time polymerase chain reaction analysis showed that FADD regulates the expression of IDE at the transcriptional level without affecting the stability of the mRNA in HepG2 cells. Following treatment with cycloheximide, the IDE protein degradation rate was found to be increased in both FADD-D primary hepatocytes and FADD-knockdown HepG2 cells. Additionally, IDE expression levels were reduced in insulin-stimulated primary hepatocytes from FADD-D mice compared to those from control mice. Moreover, FADD phosphorylation promotes nuclear translocation of FoxO1, thus inhibiting the transcriptional activity of the IDE promoter. Together, these findings imply a novel role of FADD in the reduction of protein stability and expression levels of IDE.