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당근 현탁배양세포에서 Ca2+ 과 Polyamine 이 Callose 함량에 미치는 영향
표병식(Byoung Sik Pyo),강영희(Young Hee Kang) 한국식물학회 1989 Journal of Plant Biology Vol.32 No.4
The effects of Ca^2+ on polyamines on callose contents of carrot suspension cultured cells were studied. The regeneration process of the cell wall of carrot protoplast observed through the electron microscope. Treatment of the carrot suspension cultured cells with Ca^2+ and polyamines resulted in considerable increase on callose contents at 0.1 mM of Ca^2+ and polyamines, particulary spermidine. Poly-L-lysine and poly-L-ornithine increased about 30% and 100% of callose contents than that of the control respectively, whereas verapamil and flunarizine markedly decreased the callose contents. These effects of Ca^2+ and polyamines may be ascribed to a polycationic nature on the callose contents. Futhermore, Ca^2+ and calmodulin increased the callose contents as Ca^2+ of free ion rather than as Ca^2+ -calmodulin complex. During the cultivation of the protoplast, the regeneration of the cell wall was somewhat observed on the 4th day, however, it was inhibited by verapamil. These results suggested that the promotive action of Ca^2+ and polyamines were manifested in the callose contents and the regeneration of the cell wall.
완두 발아시 Polyamine 함량 변화 및 Peroxidase Isozyme 양상
표병식(Byoung Sik Pyo),이영주(Young Ju Lee),강영희(Young Hee Kang) 한국식물학회 1993 Journal of Plant Biology Vol.36 No.4
In germinating pea, contents of endogenous polyamine in the leaf and stem were determined, and protein content, peroxidase activity and pattern of isozymes were examined in the leaf treated with polyamines. During growth of the pea for 14 days in light condition, the polyamines in leaf and stem showed the highest level at the 5th day, and were decreased rapidly at the 7th day, kept almost constant level since then. The putrescine level was relatively higher than those of spermidine and spermine, and cadaverine was also detected. On the other hand, in the leaf treated with supermine (0.01 mM) protein content increased about 250% than that of the control, the peroxidase activity increased more than 100% in spermine of 0.01 mM and 0.1 mM. In treating with putrescine of 0.1 mM the pattern of peroxidase isozyme appeared 4 new cathodic bands (pI 4.8, 5.6, 5.9 and 6.8) compared with the control, the clear cathodic bands (pI 5.6, 5.9, 6.4, and 6.6) were also observed in spermine of 0.1 mM. These results suggest that polyamines were important factor in the differentiation of pea at the early stage of germination.
당근 현탁배양 세포에서 Ca2+ 과 Polyamines 가 Cell Wall 합성에 관여하는 β - Glucan Synthetase 2 활성에 미치는 영향
표병식(Byoung Sik Pyo),강영희(Young Hee Kang) 한국식물학회 1988 Journal of Plant Biology Vol.31 No.2
The effect of Ca^2+ and polyamines on the activity of β-glucan synthetase Ⅱ(GSⅡ) related to cell wall synthesis was studied in carrot suspension cultured cells. The activity of GS Ⅱ is four times higher than that of β-glucan synthetase Ⅰ in carrot suspension cultured cells and in vitro experiment, the activity of GSⅡ was increased in response to increase in concentration of Ca^2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca^2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca^2+ and polyamines, the GSⅡ activity was high at 0.1mM of Ca^2+ and 1 mM of putrescine. Also, polycationic poly-L-lysine and poly-L-ornithine increased about 50% the GSⅡ activity than that of the control, respectively. These results may imply that Ca^2+ and polyamines were related to the enzyme activity as a polycationic nature. In addition, verapamil as the calcium channel blocker and flunarizine as an antagonist of calcium mechanism in cytoplasm decreased GSⅡ activity ramarkably, Ca^2+ and calmodulin stimulated GSⅡ activity as Ca^2+ of free ion rather than Ca^2+ calmodulin complex. The effect of 2,4-D on the GSⅡ activity in culture medum is shown to be low at 0.1㎎ per liter and GSⅡ activity increased about 30% more than that of the 0.1㎎/l at the range of 0.3-1.0㎎ per litere. Cummulative results suggest that Ca^2+ and polyfamines stimulate the cell wall synthesis by means of the enhancement of GSⅡ activity responsible for synthesizing the cell wall components.
표병식(Byoung-Sik Pyo),오영준(Young-Jun Oh),박양원(Yang-Won Park) 한국식품영양과학회 1994 한국식품영양과학회지 Vol.23 No.6
토양에서 분리한 균주 IJ-3를 포함하는 미생물이 균체외의 두유응고 효소를 분비하였다. 균주 IJ-3는 Bergey' s에 의해 Bacillus속의 균체로 확인되었다. 두유응고 효소는, pH 5.8~6.4 및 55~75℃ 이하에서 커드를 형성하였다. 두유응고 활성에 대한 최적온도는 65~75℃였고, 효소는 50℃ 이하의 범위의 온도에서 안정하였으며, pH 범위 (pH 6~7)에서 효소고유 활성의 60~100%의 안정성을 보였다. 효소의 분자량은 SDS-PAGE에서 28,000으로 추정하였다. Bacillus sp. IJ-3 효소로 커드는 부드러운 조직과 쓴맛이나 콩비린내가 없는 순수맛을 보였다. Microorganisms, including the strain IJ-3 isolated from soil, were found to secrete an extracellular soybean protein coagulating enzyme and the strain, IJ-3, was identified as Genus Bacillus according to the Bergey's manual. The enzyme coagulated protein in soymilk, thus forming a curd at pHs 5.8~6.4 and at 55~75℃. The optimum temperature for soybean protein coagulating activity was 65~75℃ and the enzyme was stable at temperature below 50℃ and was found to be stable with about 60~100% of the original activity at a with pH ranges (pH 6~7). The molecular weight of enzyme was estimated to be 28,000 by SDS-PAGE. The curd formed with the enzyme from Bacillus sp. IJ-3 has a smooth texture, and a mild taste without any bitterness or a beany flavor.
HPLC와 DPPH radical 소거능 측정 방법의 결합에 의한 약용 식물 추출물의 항산화 활성 비교
임도연(Do-Youn Im),표병식(Byoung-Sik Pyo),김선민(Sun-Min Kim),이경인(Kyoung-In Lee) 한국생명과학회 2017 생명과학회지 Vol.27 No.1
본 연구에서는 일반적으로 분리 및 분석에 가장 빈번히 사용되고 있는 C18 column과 UV 검출기가 장착된 액체크로마토그래피(HPLC)와 항산화 활성 측정에 사용되는 1, 1-diphenyl-2-picryl hydrazyl (DPPH) 라디칼 소거능 측정 방법을 결합한 HPLC-DPPH 동시 측정법의 최적화와 유용성 확인을 약용식물의 추출물을 대상으로 실시하였다. 최종적으로 적용된 HPLC-DPPH 동시 측정법의 유용성은 갈근, 건강, 유근피, 모과, 황기 등 5종 약용식물의 열수추출물과 대조군으로서 ascorbic acid의 라디칼 소거능을 측정하여 확인하였다. HPLC-DPPH 동시 측정에 앞서 추출액 중 고형분 함량을 refractometer를 사용하여 측정함으로써 추출 수율에 따른 활성 차이를 보정할 수 있도록 하였다. 갈근, 모과, 유근피 열수추출물의 라디칼 소거능이 대조군으로 사용된 ascorbic acid와 비교하여 7.77%, 4.71%, 4.19%로서 다른 열수추출물보다 상대적으로 높은 것으로 확인되었다. 이와 같은 측정법은 실제 활성 성분의 분리 및 분석에 있어서 불필요한 시간 및 시약의 낭비를 줄일 수 있는 유용한 수단이 될 수 있을 것으로 판단된다. Natural anti-oxidative compounds have important disease prevention and food preservation properties, in addition to anti-bacterial, anti-inflammation, anti-cancer, and skin whitening effects. High-performance liquid chromatography (HPLC), with an ultra vilolet (UV) detector coupled to a reverse phase C18 column and an online measurement system for 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radicals, was used to search for potent antioxidative compounds in crude extracts. The online HPLCDPPH assay was then applied to confirm antioxidative compounds in water extracts from Radix of Pueraria lobata, Rhizoma of Zingiber officinale, Fructus of Chaenomeles sinensis, Cortex of Ulmus pumila, and Radix of Astragalus membranaceus. To determine the yields of the extracts, the Brix% of each extract solution was measured using a refractometer. When the relative DPPH radical scavenging ability values of the water extracts were compared with those of a positive control (ascorbic acid), the water extracts of P. lobata, C. sinensis, and U. pumila were 7.77%, 4.71%, and 4.19%, respectively. The results suggest that this method provides a useful assay for rapid measurement of DPPH radical scavenging abilities and conformation of antioxidative compounds in natural products. Moreover, it can reduce the time spent on the separation of active compounds from natural materials, such as medicinal plants, in addition to the use of reagents for separation.