響應(yīng)面法優(yōu)化天然香料β-苯乙醇液體發(fā)酵

生物工程論文編號(hào):SW121  論文字?jǐn)?shù):10759,頁(yè)數(shù):28

目錄
中文摘要………………………………………………………………………………Ⅰ
英文摘要………………………………………………………………………………Ⅱ
目錄 …………………………………………………………………………………..Ⅲ
1. 緒論…………………………………………………………………………………1
 1.1 產(chǎn)品概述…………………………………………………..…………………1
 1.3 化學(xué)合成法………………..…………………………………………………2
 1.4 生物轉(zhuǎn)化法生產(chǎn)β-苯乙醇…………………………………..……………..2
 1.5 本課題的研究任務(wù)……………………………………………………..…....4
2. 實(shí)驗(yàn)部分…………………………………………………………………………....5
 2.1 材料…………..……………………………………………………………....5
 2.2 微生物菌種……………………………………..……………………………5
 2.3 培養(yǎng)基………………………………………..………………………………6
3. 實(shí)驗(yàn)設(shè)計(jì)和方法……………………………………………………………………6
 3.1 Plackett-Burman 法篩選影響β-苯乙醇產(chǎn)量的重要因素………..……..6
 3.2 高效液相色譜法測(cè)定發(fā)酵液中L-苯丙氨酸和β-苯乙醇………………....9
 3.3 L-苯丙氨酸和β-苯乙醇標(biāo)準(zhǔn)曲線的制作……………………………........9
 3.3.1 L-苯丙氨酸標(biāo)準(zhǔn)曲線的制作………………………………………..9
 3.4 最陡爬坡實(shí)驗(yàn) ……………………………………………………...………15
 3.5 響應(yīng)面實(shí)驗(yàn)優(yōu)化發(fā)酵條件 ……………………….………………..………16
 3.6 驗(yàn)證實(shí)驗(yàn) ………………………………………….………………..………21
4. 結(jié)果與討論 ……………………………………………….……………..………..21
5. 總結(jié)與展望 …………………………………………………………….…………21
致    謝……………………………………………………………………...………..22
參考文獻(xiàn)………………………………………………………….…………..………..23

摘要：對(duì)馬克斯克魯維酵母AS 2.1440搖瓶發(fā)酵生產(chǎn)β-苯乙醇的培養(yǎng)條件進(jìn)行了統(tǒng)計(jì)優(yōu)化。基于單因子試驗(yàn)，我采用包含有十一個(gè)變量的二水平Plackett-Burman設(shè)計(jì)來(lái)篩選出對(duì)β-苯乙醇的產(chǎn)量有重要影響的相關(guān)因素。接種量，轉(zhuǎn)速和L-苯丙氨酸濃度是具有重要影響效應(yīng)的因素。然后用最陡爬坡路徑逼近上述三個(gè)因素的最佳區(qū)域。由中心組合實(shí)驗(yàn)及響應(yīng)面分析確定了這些因素的最佳條件和它們之間的相互作用。預(yù)測(cè)結(jié)果顯示在最佳條件下(L-苯丙氨酸4.0 g/L，接種量5.7%和轉(zhuǎn)速163 rpm)，通過(guò)48 h發(fā)酵β-苯乙醇的產(chǎn)量為930 mg/l。通過(guò)實(shí)驗(yàn)驗(yàn)證，最佳條件使β-苯乙醇的產(chǎn)量從402 mg/l 提高到了926mg/l。
關(guān)鍵詞：β-苯乙醇；馬克斯克魯維酵母AS 2.1440；Plackett-Burman設(shè)計(jì)；響應(yīng)面法；優(yōu)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