본 연구에서는 효소의 활성을 증진시킬 수 있는 새로운 방법의 개발을 위하여 정제된 polygalacturonase(PGase)에 guar gum, xanthan gum, locust bean gum 등과 같은 교질물질을 첨가함으로써 효소의 활성을 특이적으로 향상시킬 수 있는 방법 조사하였다. 정제된 PGase를 0.2%의 상술한 교질물질을 함유하는 동일한 완충용액에 잘 혼합시켜 30oC에서 배양하여 상대활성을 구한 결과, guar gum 첨가 시 는 PGase의 상대활성이 105%가 증가 되었으며, xanthan gum 첨가 시는 87%, locust bean gum 첨가 시는 90%가 증가되어 상술한 교질물질들이 효소의 활성 촉진제로서의 기능이 있음을 확인할 수 있었다.
Guar gum 처리에 의해 당근 단세포 생성 반응을 수행한 결과 모든 반응시간에서 guar gum을 가했을 때가 PGase만을 가하여 단세포화 반응을 수행한 경우 보다 높은 수율을 보였으며, 단세포화 반응 2시간 경과 후에 대조군 대비 13%의 가장 높은 수율 향상을 보였다.
이상의 결과에서 PGase에 guar gum, xanthan gum, locust bean gum 등과 같은 교질물질을 첨가함으로써 효소의 활성을 특이적으로 향상시킬 수 있으며, 이를 이용한 단세포 당근의 제조에 있어서 효소반응의 수율 증진을 위한 방법으로 활용될 수 있음을 확인하였다.
본 연구는 고추가 함유한 효소들이 식품이 가공과 저장시에 어떠한 영향을 미칠 것인가를 추정하고자 pectinesterase(PE), polygalacturonase(PG), lipoxygenase(LOX) and peroxidase(POD)의 특성을 조사하였다. 그 결과는 다음과 같다. 1. PE는 pH 7.5에서 최대활성을 보였으며 pH 5.5이하에서는 활성이 거의 나타나지 않았다. NaCl과 CaCl_2첨가의 경우 각각 0.2M과 0.05M에서 최대활성을 보였으며, NaCl과 CaCl_2가 PE의 활성촉진제로 작용하였다. 2. PG는 pH 6.0에서 최대활성을 보였으며 알칼리성에서보다 산성에서 활성이 컸다. NaCl과 CaCl_2첨가의 경우 각각 0.3M과 0.2mM에서 최대활성을 보였으며 이후 점차 감소하여 이들 염을 첨가하지 않았을 때보다도 활성이 감소하였다. 3. LOX는 pH 7.0과 pH 8.5에서 최대활성을 보였다. 4. POD는 pH 6.0에서 최대활성을 보였고 대체로 약산성과 중성 사이에서 활성이 크게나타났다. NaCl과 CaCl_2 같은 염은 POD활성에 약간의 저해제로 사용되었다.
To examin ripening in peach types, cell wall contents and Polygalacturonase activity were compared in Changbang, Daegubo and Yumyung peaches. Among peach types, the hardness of Daegubo was the lowest. Yumyung peach had the highest content of alcohol-insoluble substances and Changbang peach of cell wall. The contents of total and insoluble pectic substances were little different between Changbang and Yumyung peach, while the lowest in Daegubo. Daegubo peach had the highest activity of polygalacturonase, Changbang and Yumyung peach in succession.
The object of this study was to investigate the characteristics of pectinesterase(PE), polygalacturonase(PG) and Peroxidase(POD) in Kimchi materials. The results were as follows : 1. The specific activities of PEs in Korean cabbage, Korean raddish, garlic and ginger were 200 unit/mg protein, 23.1 unit/mg protein, 0.8 unit/mg protein and 32 unit/mg protein, respectively. The optimum pHs of PEs in all materials were between 7 to 8. The concentrations of NaCl, CaCl2 which showed the highest activities of PEs were 0.2~0.3M NaCl, 50 mM CaCl2 in Korean cabbage and raddish, 0.05 M NaCl, 20 mM CaCl2 in garlic and 0.2 M NaCl, 20 mM CaCl2 in ginger. 2. The specific activities and the optimum pHs of PGs were 1.5 unit/mg protein and pH 4.5 in Korean cabbage, 1.6 unit/mg protein and pH 4.5~5.5M in Korean raddish, 0.06 unit/mg protein and pH 3.0~3.5M in garlic, and 0.06 unit/mg protein and CaCl2 in ginger. The concentrations of NaCl, CaCl2 which showed the highest activities of PGs were 0.1~0.2M NaCl and 0.15~0.2mM mM CaCl2 in all materials. 3. The specific activities and the optimum pHs of PODs in Korean cabbage, Korean raddish, garlic and ginger were 71.3 unit/mg protein ; pH 6.0. 769 unit/mg protein ; pH 5.5, 1.09 unit/mg protein ; pH 4.5 and 12.7 unit/mg protein ; pH 5.0~5.5M, respectively. POD activities were not decreased in Korean cabbage, but decreased in Korean raddish by the increase of NaCl, CaCl2 concentrations. In garlic and ginger, POD activities were a little slightly affected by the increase of NaCl, CaCl2 concentrations.
The ripening behavior of three apple cultivars, ‘Tsugaru’, ‘Hongro’ and ‘Fuji’ was distinctive and the involvement of POLYGALACTURONASE(PG) in the fruit softening process was confirmed to be ethylene dependent. Fruit softening is genetically coordinated by the action of several cell wall enzymes, including PG which depolymerizes cell wall pectin. Also, loss of firmness is associated with increasing of the ripening hormone, ethylene. In this work, climacteric ripening of three apple cultivars, Tsugaru, Hongro and Fuji, producing different ethylene levels and ripening responses, was examined. Correspondingly in Fuji, a linear and basal ethylene level was observed over the entire period of measurements, and Tsugaru and Hongro displayed a typical climacteric rise in ethylene production. Transcript accumulation of genes involved in ethylene biosynthesis (MdACS3 and MdACO1) and MdPG1 was studied in Tsugaru, Hongro and Fuji cultivars. Expression of MdACO1 transcripts was shown in all three ripened apple fruits. However, the MdACS3 and MdPG1 were transcribed differently in these cultivars. Comparing the MdPG1 of ‘Tsugaru’, ‘Hongro’ and ‘Fuji’, structural difference was discovered by genomic Southern analysis. Overall results pointed out that MdACS3 and MdPG1 play an important role in regulation of fruit ripening in apple cultivar.