木鱉果假種皮冷凍乾燥萃取物的潛在蛋白酶與脂肪酶之鑑定

Identification of Potent Bioactive Protease and Lipase from Lyophilized Aril Extract of Momordica Cochichinensis Fruit

葉偉鑫(Wily)

指導教授 ： 徐志宏

國立屏東科技大學/農學院/生物科技系/碩士(2022年)

https://doi.org/10.6346/NPUST202200461

摘要

東南亞超級水果木鱉果的假種皮富含番茄紅素和其他大量類胡蘿蔔素。 已經對苦瓜的假種皮進行了大量研究，但還沒有對酶活性或蛋白質組學進行研究。 在這項研究中，液氮被用來研磨假種皮，然後進行超聲波輔助提取和冷凍乾燥。 CAP 提取物通過水解棕櫚酸 4-硝基苯酯並在 37 °C 下孵育 20 分鐘期間產生 0.0478 U/ml 的 4-硝基苯酚來顯示脂解活性。 CAP 還通過水解酪蛋白並產生 2.18 U/ml 的酪氨酸而具有蛋白水解活性。 使用凝膠 LC-MS/MS 檢測可能的蛋白酶，該蛋白質被鑑定為類奇異果甜蛋白，它是一種與在丹氏奇異球菌中發現的奇異果甜蛋白相關的半胱氨酸蛋白酶。 進行初步實驗以確定 CAP 對正常人真皮成纖維細胞 (HFC) (CG1476) 遷移的影響。

關鍵字

木鱉果 ； 蛋白酶 ； 假種皮 ； 脂肪酶 ； 酵素

並列摘要

The arils of the extraordinary Southeast Asian superfruit Momordica cochinchinensis are rich with lycopene and other enormous carotenoids. Numerous investigations have been done on the aril of Momordica cochinchinensis, but no research has been conducted on enzymatic activity or proteomics. In this investigation, liquid nitrogen was used to grind the aril, followed by ultrasound-assisted extraction and freeze-drying. CAP extracts revealed lipolytic activity by hydrolyzing 4-nitrophenyl palmitate and generating 0.0478 U/ml of 4-nitrophenol during a 20-minute incubation at 37 °C. CAP also contains proteolytic activity by hydrolyzing casein and generating 2.18 U/ml of tyrosine. Using gel LC-MS/MS to detect the probable protease enzyme, the protein was identified as a thaumatin-like protein, which is a cysteine protease associated with the thaumatin discovered in Thaumatococcus daniellii. A preliminary experiment was undertaken to determine the effect of CAP on the migration of normal human dermal fibroblasts (HFCs) (CG1476).

並列關鍵字

Gac fruit ； Protease ； Aril ； Lipase ； Enzyme

參考文獻

Addis, R., Cruciani, S., Santaniello, S., Bellu, E., Sarais, G., Ventura, C., ... & Pintore, G. (2020). Fibroblast proliferation and migration in wound healing by phytochemicals: Evidence for a novel synergic outcome. International Journal of Medical Sciences, 17(8), 1030-1042.

Ahuja, I. de, Vos RC, Bones AM, Hall RD. 2010. Plant molecular stress responses face climate change. Trends in Plant Science, 15, 664-674.

Al-Whaibi, M. H. (2011). Plant heat-shock proteins: a mini review. Journal of King Saud University-Science, 23(2), 139-150.

Amarowicz, R., Pegg, R. B., Rahimi-Moghaddam, P., Barl, B., & Weil, J. A. (2004). Free-radical scavenging capacity and antioxidant activity of selected plant species from the Canadian prairies. Food chemistry, 84(4), 551-562.

Aoki, H., Kieu, N. T. M., Kuze, N., Tomisaka, K., & Chuyen, N. V. (2002). Carotenoid pigments in GAC fruit (Momordica cochinchinensis SPRENG). Bioscience, biotechnology, and biochemistry, 66(11), 2479-2482.4