• 한국동물학회:학술대회논문집
• /
• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
• /
• pp.201.2-201
• /
• 1996

No Abstract, See Full Text

• Animal Bioscience
• /
• 제34권3_spc호
• /
• pp.463-470
• /
• 2021

Objective: This experiment was conducted to find out the immunological effects of wheat phytase when long-chain inorganic polyphosphate (polyP) treated with wheat phytase was added to a macrophage cell line, Raw 264.7, when compared to intact long-chain polyP. Methods: Nitric oxide (NO) production of Raw 264.7 cells exposed to P700, a long-chain polyP with an average of 1,150 phosphate residues, treated with or without wheat phytase, was measured by Griess method. Phagocytosis assay of P700 treated with or without phytase in Raw 264.7 cells was investigated using neutral red uptake. The secretion of tumor necrosis factor α (TNF-α) by Raw 264.7 cells with wheat phytase-treated P700 compared to intact P700 was observed by using Mouse TNF-α enzyme-linked immunosorbent assay kit. Results: P700 treated with wheat phytase effectively increased NO production of Raw 264.7 cells by 172% when compared with intact P700 at 12 h exposure. At 5 mM of P700 concentration, wheat phytase promoted NO production of macrophages most strongly. P700, treated with wheat phytase, stimulated phagocytosis in macrophages at 12 h exposure by about 1.7-fold compared to intact P700. In addition, P700 treated with wheat phytase effectively increased in vitro phagocytic activity of Raw 264.7 cells at a concentration above 5 mM when compared to intact P700. P700 dephosphorylated by wheat phytase increased the release of TNF-α from Raw 264.7 cells by 143% over that from intact P700 after 6 h exposure. At the concentration of 50 μM P700, wheat phytase increased the secretion of cytokine, TNF-α, by 124% over that from intact P700. Conclusion: In animal husbandry, wheat phytase can mitigate the long-chain polyP causing damage by improving the immune capabilities of macrophages in the host. Thus, wheat phytase has potential as an immunological modulator and future feed additive for regulating immune responses caused by inflammation induced by long-chain polyP from bacterial infection.

• Animal Bioscience
• /
• 제35권6호
• /
• pp.892-901
• /
• 2022

Objective: This study was performed to investigate the potential effect of wheat phytase on long-chain inorganic polyphosphate (polyP)-mediated interleukin 8 (IL-8) signaling in an intestinal epithelial cell line, HT-29 cells. Methods: Cell viability and the release of the pro-inflammatory cytokine IL-8 in HT-29 cells exposed to polyP1150 (average of 1,150 phosphate residues) treated with or without wheat phytase were measured by the EZ-CYTOX kit and the IL-8 ELISA kit, respectively. Also, the activation of cellular inflammatory factors NF-κB and MAPK (p38 and ERK 1/2) in HT-29 cells was investigated using ELISA kits. Results: PolyP1150 negatively affected the viability of HT-29 cells in a dose-dependent manner. However, 100 mM polyP1150 dephosphorylated by wheat phytase increased cell viability by 1.4-fold over that of the intact substrate. Moreover, the 24 h exposure of cells to enzyme-treated 50 mM polyP1150 reduced the secretion of IL-8 and the activation of NF-κB by 9% and 19%, respectively, compared to the intact substrate. PolyP1150 (25 and 50 mM) dephosphorylated by the enzyme induced the activation of p38 MAPK via phosphorylation to 2.3 and 1.4-fold, respectively, compared to intact substrate, even though it had little effect on the expression of ERK 1/2 via phosphorylation. Conclusion: Wheat phytase could attenuate polyP1150-induced IL-8 release in HT-29 cells through NF-κB, independent of MAP kinases p38 and ERK. Thus, wheat phytase may alleviate inflammatory responses including hypercytokinemia caused by bacterial polyP infection in animals. Therefore, wheat phytase has the potential as an anti-inflammatory therapeutic supplement in animal husbandry.

• 미생물학회지
• /
• 제28권2호
• /
• pp.134-144
• /
• 1990

In order to examine that what kind of system correlated with cadmium detoxification mechanism in Klebsiella aerogenes ATCC 10031, we tried to investigate the effect of phosphate upon the detoxification and also elucidate whether the cadmium phosphate and/or polymeric Cd-Pi complex is formed actually in cell or not. As the results, it was shown that growing pattern had long lag adaptive phase of 12 hr to 24 hr, at the concentrations of 0.02 mM and 0.08 mM cadmium, respectively. Cadmium was accumulated more highly in the fraction of cell wall and membrane than in those of cytoplasm. In case of phosphate starving cells added cadmium, inorganic polyphosphate system was primarily correlated with Cd-detoxification during the lag phase for the accommodation to cadmium, on the other hand, Cd:Sulfide complex system secondarily correlated it during the stationary phase. These results implied that polyphosphate system and Cd:sulfide complex system, these two systems were operated compensatively each other. Considering the results obsdrved with EM and examined tha changes of sulfide and polyphosphate amount, it was reflected that Cd:S complex was located at the cell surface. In the results of $in-vivo^{31}$P NMR spectra in the cells with cadmium pressure, several phosphate signals arose newly from the polyphosphate region with moving chemical shift of it. This phinomenon strongly implied the actual existence of Dd:Pi comples and /or Cd:poly-P complex in the cell and also the cellular compartmentalization of cadmium detoxifying mechanism.

• 한국동물학회:학술대회논문집
• /
• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
• /
• pp.93-93
• /
• 1995

No Abstract, See Full Text

• 한국균학회지
• /
• 제23권1호통권72호
• /
• pp.71-79
• /
• 1995

Rhizopus oryzae의 카드뮴 적응 및 해독기작과 이에 관련된 세포내 생리 생화학적 변화를 조사하였다. R. oryzae는 카드뮴 첨가농도가 증가함에 따라 거의 비례적으로 잠복기가 길어졌다. 카드뮴의 해독에 관련하는 system은 카드뮴 첨가 배양시에만 적응기 중에 특이하게 유도, 생성되어져 카드뮴 결합단백질의 성격을 지닐 것으로 추정되는 2종류의 단백질과 동일한 시기, 조건하에서 유의하게 증가되는 무기인산중합체 pool에 의하여 1차적으로 운용되어질 것으로 사료되며, 2차적, 보완적 system은 ACPase의 derepression 그리고 phosphatidyl serine의 합성증가 등의 방법을 통하여 운용될 것으로 생각된다. 또한 총지질화합물의 조성변화 등과 같은 다양한 생리 생화학적 물질대사 경로의 변화를 일으켰다. 이러한 변화의 총화로 크게는 형태적 변화까지 일어나 카드뮴 적응 및 해독에 필요한 성장형태로서 여러 가지 산물 생성에 유리한 dispersed filamentous type을 취하였다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제33권1호
• /
• pp.127-131
• /
• 2020

Objective: This study was conducted to determine catalytic properties of wheat phytase with exopolyphosphatase activity toward medium-chain and long-chain inorganic polyphosphate (polyP) substrates for comparative purpose. Methods: Exopolyphosphatase assay of wheat phytase toward polyP75 (medium-chain polyP with average 75 phosphate residues) and polyP1150 (long-chain polyP with average 1150 phosphate residues) was performed at pH 5.2 and pH 7.5. Its activity toward these substrates was investigated in the presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or ethylenediaminetetraacetic acid (EDTA). Michaelis constant (K_m) and maximum reaction velocity (V_max) were determined from Lineweaver-Burk plot with polyP75 or polyP1150. Monophosphate esterase activity toward p-nitrophenyl phosphate (pNPP) was assayed in the presence of polyP75 or polyP1150. Results: Wheat phytase dephosphorylated polyP75 and polyP1150 at pH 7.5 more effectively than that at pH 5.2. Its exopolyphosphatase activity toward polyP75 at pH 5.2 was 1.4-fold higher than that toward polyP1150 whereas its activity toward polyP75 at pH 7.5 was 1.4-fold lower than that toward polyP1150. Regarding enzyme kinetics, K_m for polyP75 was 1.4-fold lower than that for polyP1150 while V_max for polyP1150 was 2-fold higher than that for polyP75. The presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or EDTA (1 or 5 mM) exhibited no inhibitory effect on its activity toward polyP75. Its activity toward polyP1150 was inhibited by 1 mM of Ni²⁺ or Co²⁺ and 5 mM of Ni²⁺, Co²⁺, or Mg²⁺. Ni²⁺ inhibited its activity toward polyP1150 the most strongly among tested additives. Both polyP75 and polyP1150 inhibited the monophosphate esterase activity of wheat phytase toward pNPP in a dose-dependent manner. Conclusion: Wheat phytase with an unexpected exopolyphosphatase activity has potential as a therapeutic tool and a next-generational feed additive for controlling long-chain polyP-induced inappropriate inflammation from Campylobacter jejuni and Salmonella typhimurium infection in public health and animal husbandry.

• 한국균학회지
• /
• 제10권2호
• /
• pp.57-65
• /
• 1982

경정곰팡이(Aspergillus niger)의 포자형성(胞子形成)을 액침배양법(液浸培養法)과 동조적방법(同調的方法)으로 여행(旅行)하면서 인산화합물(燐酸化合物), 고인산화(高燐酸化)뉴크레오티드, 및 RNA(핵산)(核酸)의 동태(動態)를 연구(硏究)하였다. 결과(結果)는 다음과 같이 요약(要約)된다. 1. 곰팡이의 포자형성시(胞子形成時)에 유기인산화합물(有機燐酸化合物)의 양(量)이 증가(增加)하였으며, 무기(無機) Polyphosphate의 양(量)은 반대(反對)로 감소(減少)하였다. 2. 무기인산양(無機燐酸量)은 변동(變動)하지 않았으며, 총인산화합물(總燐酸化合物)의 양(量)은 초기균사생장시(初期菌絲生長時)에는 감소(減少)하고 포자낭병시기(胞子囊柄時期)부터는 변동(變動)하지 않았다. 3. RNA핵산구획(核酸區劃)에 있어서 orcinol 반응양성구(反應陽性區)의 흡광도(吸光度)가 급격(急激)히 증가(增加)하였다. 그러나 UV흡광도(吸光度)는 완만하게 증가(增加)하였다. 4. 포자낭병(胞子囊柄)및 포자형성시기(胞子形成時期)에 guanosinetetraphosphate가 검출현(檢出現) 되었다. 5. 고분자양성(高分子量性) RNA (rRNA분획(分劃))의 polyacrylamide gel 전기영동(電氣泳動)을 2.6% gel로서 시행(施行)한 바 polyphosphate와 RNA의 결합물(結合物)이 상당양(相當量) 존재(存在)함을 알았다. 이상(以上)의 결과(結果)로 보아 검정곰팡이의 분화과정(分化過程)에 있어서 고인산화(高燐酸化) RNA 화합물(化合物)의 존재(存在)가 확인(確認)되었으며, 이 화합물(化合物) 가운데 guanosine tetraphosphate (G4P)의 존재(存在)가 진핵미생물(眞核微生物)인 곰팡이에서도 검출(檢出)되었음을 지적(指摘)한다.

• 미생물학회지
• /
• 제43권1호
• /
• pp.72-75
• /
• 2007

Escherichia coli에서 RNA 분해와 가공에 있어서 중심적인 역할을 하는 RNase E의 동족체 단백질인 Streptomyces coelicolor RNase ES의 결합 단백질을 항체침전을 이용하여 탐색하였다. 무기인산을 이용하는 polyphosphate kinase와 리보핵산외부분해효소인 polynucleotide phophorylase의 동족체인 GPSI가 RNase ES와 함께 침전되는 것을 확인하였으며, 이는S. coelicolor에도 E. coli RNase E를 매개로 구성되는 다단백질복합체인 degradosome이 RNase ES에 의해 형성될 수 있음을 암시한다. 계통적으로 멀리 떨어진 이 두 세균에서 정제된 polynucleotide phosphorylase 동족체는 시험관에서의 RNA 분해 특성이 서로 유사함을 보였다. 이러한 실험 결과는 RNase ES가 E. coli degradosome과 유사한 기능과 구조를 가진 단백질 복합체를 형성함을 시사한다.

• Applied Biological Chemistry
• /
• 제43권3호
• /
• pp.163-168
• /
• 2000

토양 및 수계에 집적, 유입되어 있는 과다한 인산의 제거에 이용 가능한 미생물 자원을 확보하기 위하여 활성오니에서 분리된 Acinetobacter lwoffi PO8의 배양조건 및 외부환경조건에 따른 인산흡수 양상을 조사하였다. 배양액 내 초기 pH가 $7.5{\sim}8.5$ 범위에서 균생육과 인산흡수율이 가장 높았으며, 탄소원으로 glycerol 및 arabinose을 첨가하였을 경우 각각 약 93 및 91%의 높은 인산 흡수율을 보였다. 질소원 형태에 따른 인산흡수양상은 아미노태 보다 암모늄염이 효과적이었으며, 특히, $NH_4NO_3$,와 $(NH_4)_2SO_4$가 각각 95와 96%의 인산흡수율을 나타냈다. 금속이온 중에서 $Co^{2+}$ 첨가 시 균생육이 제해되었으나, 이외의 다른 금속이온은 균의 성장 및 인산흡수에 큰영향을 미치지 않았다. 아미노산 중 arginine, methionine 및 lysine 등은 아미노산을 첨가하지 않은 경우보다 $10{\sim}20%$ 더 높은 인산흡수율을 나타냈었으며, 이때 배양기간 중 glucose의 공급으로 배지 중 잔존 인산이 완전히 제거되었다. 또한 $^{32}P$를 사용하여 균의 인산 분포를 조사한 결과로 세포내 흡수된 인산은 대부분 세포의 원형질 내에 polyphosphate의 형태로 분포되어 있었다.