球形交聯(lián)殼聚糖固定化風(fēng)味蛋白酶的制備及特性研究

摘要：研究了球形交聯(lián)殼聚糖固定化風(fēng)味蛋白酶的制備及其特性，以殼聚糖為載體，戊二醛為交聯(lián)劑制備固定化風(fēng)味蛋白酶，測(cè)定了不同戊二醛濃度、交聯(lián)時(shí)間、給酶量、固定化時(shí)間、固定化溫度及固定化pH對(duì)固定化風(fēng)味蛋白酶酶活性的影響。結(jié)果表明：固定化反應(yīng)的最佳條件為戊二醛濃度0.5%，交聯(lián)時(shí)間6h，給酶量40mg/g載體 ,固定化時(shí)間10h，固定化pH5.5，固定化溫度35℃。在此條件下制備的固定化風(fēng)味蛋白酶機(jī)械強(qiáng)度最大，酶活回收率最高，固定化酶的pH穩(wěn)定性與溫度穩(wěn)定性均得到提高，連續(xù)使用11次后固定化酶活力還剩余56.7%。
關(guān)鍵詞：風(fēng)味蛋白酶，固定化，球形交聯(lián)殼聚糖，特性

Study on Preparation and Properties of Immobilized FlavourzymeonGlobular Cross-linked Chitosan
Abstract:The preparation and properties of immobilized flavourzymeon globular cross-linked chitosanwere investigated.Flavourzyme has been immobilized on chitosan with glutaraldehyde as crosslinking agent. The effects on the activity of immobilized flavourzyme, such as crosslinking time, concentration of glutaraldehyde,flavourzyme dosage, reaction time, reaction temperature and reaction pH were studied. The results showed that the optimal conditions of immobilization: the optimal concentration of glutaraldehyde was 0.5%, the optimal flavourzyme dosage was 40mg/g carrier, the optimal reaction time, temperature and pH were 6h, 35℃ and 5.5 respectively. Under the optimal conditions, the immobilized flavourzyme have the best mechanical strength and the highest enzyme recovery, the pH and temperature stability were considerably improved. The recovery activity of immobilized flavourzyme was more than 66.1% after being repeated eleven times.

Key words:flavourzyme; immobilization; globular cross-linked chitosan; properties

目錄
中文摘要.............................................................................................................................1中文關(guān)鍵詞.........................................................................................................................1英文摘要............................................................................................................................ 1英文關(guān)鍵詞.........................................................................................................................1 1前言..................................................................................................................................2
1.1殼聚糖的應(yīng)用………………………………………………………………………...2
1.2固定化酶的主要方法……………………………………………………………...…2
1.3風(fēng)味蛋白酶………………………………………………………………………...…4
2 材料與方法.....................................................................................................................5
2.1材料...............................................................................................................................5
2.2儀器...............................................................................................................................5
2.3方法...............................................................................................................................5
3結(jié)果與分析......................................................................................................................5
3.1.殼聚糖固定化風(fēng)味酶的最佳條件選則.......................................................................5
3.1.1NaOH濃度和殼聚糖濃度的影響.............................................................................6
3.1.2戊二醛濃度和交聯(lián)時(shí)間的影響................................................................................6
3.1.3給酶量對(duì)固定化酶活性的影響................................................................................7
3.1.4 固定化反應(yīng)時(shí)間的影響...........................................................................................8
3.1.5 固定化體系溫度和ph的影響.................................................................................9
3.2 球形交聯(lián)殼蛋白固定化風(fēng)味酶的特性....................................................................10
3.2.1固定化酶的最佳反應(yīng)ph.........................................................................................11
3.2.2固定化酶的最佳反應(yīng)溫度......................................................................................11
3.2.3固定化酶的儲(chǔ)存穩(wěn)定性..........................................................................................12
3.2.4固定化酶的操作穩(wěn)定性..........................................................................................12
4結(jié)論................................................................................................................................14
參考文獻(xiàn)...........................................................................................................................15
致 謝......................................................................................