Purification, preparation and activity analysis of rice bran lipase

• Main Authors: Chong-You Li, 李重佑
• Other Authors: Chen, Wen-Lung
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/2e68j4

碩士 === 國立嘉義大學 === 應用化學系研究所 === 106 === Rice bran is a byproduct of the rice milling process. It is the epidermis of rice and is rich in many valuable ingredients such as carbohydrates, proteins, oils, and other phytochemicals. Rice bran is usually used as animal feed, and some are discarded directly after rice milling. Therefore, rice bran is a low economic value substance. In this work, we want to use low-cost rice bran to obtain functional or economical product through the experiments. This paper will focus on the preparation and purification of rice bran protein and investigate the activity of lipase in rice bran protein. Lipase (triacylglycerol acylhydrolase EC 3. 1. 1. 3) is classified as a hydrolyzed enzyme, which is widely used in products or industrial production processes, such as detergents, cosmetic, dairy industry, hydrolysis reaction and synthesis of oil esters ... and so on. In this work, we use the rice bran as a material to prepare the rice bran protein, and investigate changes in enzyme activity during separation, and the hydrolytically active lipase was isolated and purified. After the rice bran was extracted through the buffer solution and then partitioned by the saturation of ammonium sulfate (20-80%), the precipitated protein was removed and through the dialysis to remove the small molecular weight substances, and the activity of hydrolysis was measured by the hydrolysis reaction of para-nitrophenyl butyrate. And investigate the effect of ammonium sulfate and separation steps on activity. Then followed by further purification by gel filtration chromatography (Sephacryl-S200HR, Superdex 75) and ion exchange chromatography (DEAE-Sepharose Fast Flow), and the fractions with hydrolytic activity were collected and analyzed by electrophoresis.