固定化脂肪酶催化丁酸乙酯的合成

生物工程論文編號(hào):SW071  論文字?jǐn)?shù):9939,頁(yè)數(shù):26

目  錄
中文摘要 I
英文摘要 II
目  錄 III
1   緒論 1
1.1 脂肪酶固定化的研究概況 1
1.2 非水相中固定化脂肪酶催化性能的研究概況 4
2   實(shí)驗(yàn)部分 5
2.1  實(shí)驗(yàn)試劑及設(shè)備 5
2.2  實(shí)驗(yàn)方法及步驟 6
3  實(shí)驗(yàn)結(jié)果與討論 8
3.1 標(biāo)準(zhǔn)曲線的繪制 8
3.2 氣相圖譜分析 8
3.3   反應(yīng)進(jìn)程 9
3.4   固定化酶加入量對(duì)酯合成反應(yīng)的影響 9
3.5   酶加載量對(duì)酶活力的影響 10
3.6   底物濃度及摩爾比對(duì)酯合成反應(yīng)的影響 11
3.7   游離酶含水量對(duì)酯合成反應(yīng)的影響 12
3.8   表面活性劑對(duì)游離酶酶活的影響 13
3.9   固定化脂肪酶的儲(chǔ)存穩(wěn)定性 14
3.10  固定化脂肪酶的溫度穩(wěn)定性 15
4   總結(jié)與展望 16
致   謝 17
參考文獻(xiàn) 18
附錄 19

摘 要：近年來(lái)，隨著綠色化學(xué)的興起和對(duì)節(jié)能、環(huán)保的重視，在非水相反應(yīng)體系中使用固定化酶不但能在一定程度上減輕溶劑對(duì)酶的失活作用，而且固定化酶的分散更為均勻，避免了酶的聚集，易于處理。本實(shí)驗(yàn)以毛巾為載體，用戊二醛進(jìn)行吸附交聯(lián)固定脂肪酶，并利用固定化脂肪酶在非水相中催化正丁酸和乙醇的合成反應(yīng)。研究了固定化過(guò)程中載體質(zhì)量、酶加載量，反應(yīng)體系中的時(shí)間、底物濃度及摩爾比、含水量、表面活性劑、儲(chǔ)存和溫度穩(wěn)定性等因素對(duì)催化丁酸乙酯反應(yīng)的影響規(guī)律，確定最優(yōu)條件：在0.6g載體上固定脂肪酶0.1g／10ml為宜，在反應(yīng)體系中乙醇和丁酸最適比例約為1:2，游離酶最適含水量為O.5％。表面活性劑中Tween60對(duì)酶活性的提升效果最為明顯，另外，在時(shí)間、穩(wěn)定性研究中對(duì)底物和產(chǎn)物的趨勢(shì)亦作了討論。

關(guān)鍵詞：脂肪酶；固定化；酯催化；丁酸乙酯

Catalytic synthesis of ethyl butyrate by immobilized lipase

Abstract: In recent years, with the rise of green chemistry and energy-saving, environmental protection, enzyme reuse has become an important topic of the biological engineering, enzyme immobilization is one of the methods to solve this problem, In no water system, immobilized enzyme system not only can alleviate deactivation function of solvent for enzyme, but also can scatter more evenly in the system, which makes it easy to handle. This experiment with towel for the carrier, with glutaraldehyde fixation lipase on adsorption crosslinking and use in the immobilized lipase studied.it butyric acid catalytic synthesis of ethanol and positive reaction. Study the immobilized enzyme process quality, the carrier, the reaction system load time, substrate concentration and mole ratio, moisture content, surfactants, storage and temperature stability of catalytic ethyl butyrate reaction and determine the influence law of the optimal conditions. At 0.6 g carrier fixed lipase 0.1 g / 10ml advisable, in the reaction system in ethanol and butyric acid optimal ratio of 1:2 for the optimal water free enzymes, surfactants, 0.5% of enzyme activity in the ascension Tween60 most obvious effect, in addition, in time, stability studies on substrates and product trend is also discussed.

Keywords：Lipase; immobilized; ester catalytic; ethyl butyrate