响应面法优化天然香料β-苯乙醇液体发酵

论文编号:SW121  论文字数:10759,页数:28

摘要：对马克斯克鲁维酵母AS 2.1440摇瓶发酵生产β-苯乙醇的培养条件进行了统计优化。基于单因子试验，我采用包含有十一个变量的二水平Plackett-Burman设计来筛选出对β-苯乙醇的产量有重要影响的相关因素。接种量，转速和L-苯丙氨酸浓度是具有重要影响效应的因素。然后用最陡爬坡路径逼近上述三个因素的最佳区域。由中心组合实验及响应面分析确定了这些因素的最佳条件和它们之间的相互作用。预测结果显示在最佳条件下(L-苯丙氨酸4.0 g/L，接种量5.7%和转速163 rpm)，通过48 h发酵β-苯乙醇的产量为930 mg/l。通过实验验证，最佳条件使β-苯乙醇的产量从402 mg/l 提高到了926mg/l。
关键词：β-苯乙醇；马克斯克鲁维酵母AS 2.1440；Plackett-Burman设计；响应面法；优化
 
Optimization of Submerged Fermentation of β-phenylethanol Production by Kluyveromyces marxious with Response Surface Methodology

Abstract: The cultivation conditions for β-Phenylethanol (β-PE) production using Kluyveromyces marxious AS 2.1440 were optimized statistically in shake flask experiments. Based on a single factor experiment design, we implemented the two-level Plackett–Burman (PB) design with 11 variables to screen related factors that significantly influence β-PE production. Inoculum, speed and L-Phenylalanine concentration were identified as the most important significant factors. Subsequently, the path of steepest ascent was used to approach the optimal region of the above three factors. Finally, Response surface methodology based on the Center Composite Design and response surface analysis was applied to determine these factors’ optimal levels and their mutual interactions for β-PE production. The predicted results showed that 930 mg/l of β-PE production was obtained after a 48-h fermentation period, with optimal condition of L-Phe (4.0g/L), inoculum (5.7%), and speed (163 rpm) in the flask fermentations. The optimized conditions allowed β-PE yield to be increased from 402 mg/l to 926 mg/l after verification experiments test.
Keywords: β-phenylethanol; Kluyveromyces marxious AS2.1440; Plackett-Burman design; response surface method; optimization

目录
中文摘要………………………………………………………………………………Ⅰ
英文摘要………………………………………………………………………………Ⅱ
目录 …………………………………………………………………………………..Ⅲ
1. 绪论…………………………………………………………………………………1
 1.1 产品概述…………………………………………………..…………………1
 1.3 化学合成法………………..…………………………………………………2
 1.4 生物转化法生产β-苯乙醇…………………………………..……………..2
 1.5 本课题的研究任务……………………………………………………..…....4
2. 实验部分…………………………………………………………………………....5
 2.1 材料…………..……………………………………………………………....5
 2.2 微生物菌种……………………………………..……………………………5
 2.3 培养基………………………………………..………………………………6
3. 实验设计和方法……………………………………………………………………6
 3.1 Plackett-Burman 法筛选影响β-苯乙醇产量的重要因素………..……..6
 3.2 高效液相色谱法测定发酵液中L-苯丙氨酸和β-苯乙醇………………....9
 3.3 L-苯丙氨酸和β-苯乙醇标准曲线的制作……………………………........9
 3.3.1 L-苯丙氨酸标准曲线的制作………………………………………..9
 3.4 最陡爬坡实验 ……………………………………………………...………15
 3.5 响应面实验优化发酵条件 ……………………….………………..………16
 3.6 验证实验 ………………………………………….………………..………21
4. 结果与讨论 ……………………………………………….……………..………..21
5. 总结与展望 …………………………………………………………….…………21
致    谢……………………………………………………………………...………..22
参考文献………………………………………………………….…………..………..23

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