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| 皱褶假丝酵母脂肪酶催化水解大豆油工艺优化及机制研究 |
| Process optimization and mechanism of hydrolysis of soybean oil catalyzed by lipase from Candida rugosa |
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| DOI： |
| 中文关键词: 皱褶假丝酵母脂肪酶 大豆油 脂肪酸 水解 机制 |
| 英文关键词:lipase from Candida rugosa soybean oil fatty acid hydrolysis mechanism |
| 基金项目: |
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| 中文摘要: |
| 为了降低脂肪酶催化水解植物油脂的应用成本，以皱褶假丝酵母脂肪酶（CRL）为催化用酶，对比了CRL催化水解不同植物油脂的效果，同时以大豆油为研究对象，采用单因素实验考察pH、反应温度、反应时间、水油体积比和酶用量对水解率的影响，并运用响应面法优化工艺条件，进而对CRL催化甘油三酯水解的机制进行了总结。结果表明：CRL催化水解C18脂肪酸甘油酯的选择性较高；CRL催化大豆油水解的最优工艺条件为反应温度39.5℃、pH 6.7、水油体积比1∶2.2、酶用量0.6%(以大豆油质量计)，在此条件下反应2 h水解率可达77.21%，反应12 h水解率可达96.33%;CRL催化水解甘油三酯的反应机制主要为亲核催化三联体中的丝氨酸对甘油三酯的羰基进行亲核反应以夺取脂肪酸前体，然后水分子对该前体进行亲核反应释放脂肪酸。CRL能够在温和条件下高效催化植物油脂生产脂肪酸，是一种具有广阔应用前景的生物催化剂。 |
| 英文摘要: |
| In order to reduce the application cost of lipase catalyzed hydrolysis of vegetable oil, the lipase from Candida rugosa(CRL) was used as a catalyst, and the effect of hydrolysis of different vegetable oils with CRL was compared. Selecting soybean oil as the research object, the influence of pH, reaction temperature, reaction time, volume ratio of water to oil and dosage of enzyme on hydrolysis rate was investigated by single factor experiment, then the response surface methodology was applied to optimize the process conditions. The results indicated that the selectivity of hydrolysis of C18 fatty acid glyceride catalyzed by CRL was higher. The optimal process conditions of hydrolysis of soybean oil catalyzed by CRL were obtained as follows: reaction temperature 39.5℃, pH 6.7, volume ratio of water to oil 1∶2.2, and enzyme dosage 0.6% (based on soybean oil mass). Under these conditions, the hydrolysis rate achieved 77.21% within 2 h and 96.33% within 12 h. The main reaction mechanism of CRL catalyzed hydrolysis of triglycerides was serine in the nucleophilic catalytic triad reacted nucleophilically with the carbonyl group of triglyceride to seize the fatty acid precursor, and then water molecule reacted nucleophilically with the precursor to release fatty acid. CRL can efficiently catalyze the production of fatty acids from vegetable oils under mild conditions, and it is a biocatalyst with broad application prospects. |
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