臺灣博碩士論文加值系統

苯丙胺酸為帶苯環基的胺基酸，是人體八種必需胺基酸之一，即是必
須由外界飲食中攝取來補充，人體內部無法自行製造。除了當作阿斯巴甜
的原料之外也有在食品、醫藥及飼料添加劑等多種用途。生產方式有蛋白
質水解法、有機化學合成法、微生物直接醱酵法及酵素轉化法四種。酵素
轉化法是將微生物醱酵生長後，將微生物取出，再加入適當的反應先驅物
，藉由微生物體內的某些物酵素將這些先驅物轉化為苯丙胺酸，些種方式
可以有較高的轉化率及副產物少，易於回收純化等優點，未來市場的競爭
潛力大，是最被看好的苯丙胺酸生產方式。 本論文研究之目的在探討
乳化液膜包覆系統對L-苯丙胺酸生產酵素活性之影響。實驗系統分別採用
氨基轉移酵素( AAT ) 與苯丙胺酸氨解酵素( PAL )兩種不同類型之酵素
進行比較，製造AAT酵素之微生物菌種細胞( E. coli， E. coli 改質細
胞 VJS632/pTYR100，VJS676/pASP400 ) 及製造PAL酵素之酵母菌種細胞(
R. toruloides )，經過培養、發酵、離心、分離後，分別製成細胞懸浮
液與酵素溶液，再利用乳化液膜固定化技術分別予以包覆，進行酵素催化
反應。實驗方法採用田口式實驗計畫法進行規劃，分別探討油相組成對
AAT及PAL兩種酵素，在不同操作系統下對其活性之影響，並建最佳之乳化
液膜操作條件。

The objective of this research is to investigate the
effect of emulsion liquid membrane ( ELM ) encapsulation system
on the activity of L-phenylalanine-produced enzymes. Two kinds
of enzymes , aromatic amino acid amino transferase ( AAT ) and
L-phenylalanine ammonia-lyase(PAL)，are adopted in the study .
The enzymes o f AAT and PAL are biologically synthesized form
fermentation of E.coli , genetically - modified E.coli (VJS632/
pTYR100,VJS676/pASP400 ) and R. toruloides ，respectively. After
cultivation of enzyme of enzyme-producing strains, the
intracellular enzymes are harvested into two forms : whole
cell suspension and crude enzyme solution , which are
subsequently encapsulated by ELM to perform the enzymatic
reaction. Taguchi''s experimental design is adopted to
discussed the oil membrane effect on the enzyme activity in
different operating systems. The results compiled form this
research are expected to be a reference for optimzing the
ELM encapsulation system.