基于羧基载体LX-1000IDA的脂肪酶固定化研究

doi:10.16088/j.issn.1001-6600.2020.06.011

Abstract

Abstract: This paper used carbon diamine EDC as the activator of the carboxyl carrier LX-1000IDA, and the marine lipase was fixed on the carboxyl carrier through the surface amino resin of lipase to generate the fixed marine lipase IDA-LIPASE. To achieve the best immobilization effect, conditions such as the order of addition of carriers, enzymes and crosslinking agents, buffer concentration, temperature, pH, carrier quantity, time and EDC concentration were optimized and screened. After using single-factor experiment, Plackett Burman experiment, and orthogonal experiment, the optimized conditions of immobilization were determined as follows: pH 4.5, carrier amount 0.2 g, time 6h, temperature 25 ℃, 0.6% EDC, The immobilized enzyme was prepared with enzyme activity of 114 U/g. After 10 times of expanded immobilization, the activity of the immobilized IDA- LIPASE reached 210 U/g. After comparing the prepared immobilized lipase with free enzyme, it was found that: the optimal reaction temperature increased by 10 ℃, the optimal reaction pH shifted 0.5 to the alkaline direction, and the reusability was better. About 40% of the enzyme activity was still left after the 7th usage of immobilized catalysis, the immobilize lipase exhibited excellent thermal stability. In addition, the immobilized lipase remained 84.6% enzyme activity after preservation at 4 ℃ for 30 d and exhibited good storage stability. Compared with free enzyme, the immobilized enzyme IDA-LIPASE was more suitable for industrial production environment and possessed the potential and value of practical production.

Key words: marine lipase, immobilization, carboxyl carrier, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, enzymatic property

CLC Number:

Q814.2

ZHU Heng, ZHANG Jifu, ZHANG Yun, HU Yunfeng. Immobilization Method of Lipase Based on Carboxyl Carrier LX-1000IDA[J].Journal of Guangxi Normal University(Natural Science Edition), 2020, 38(6): 88-101.

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Immobilization Method of Lipase Based on Carboxyl Carrier LX-1000IDA

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