木質纖維素中的葡萄糖與木糖可作為微生物醱酵的基質，然而麵包酵母與木質醋酸菌均無法有效地利用木糖生產酒精與細菌纖維素；藉由木糖異構酶 (xylose isomerase) 將木糖進行異構化反應，其產物木酮糖 (xylulose) 卻可被有效地利用。以同步異構化醱酵(simultaneous isomerization and fermentation；SIF ) 方式培養，可提高木酮糖之轉換率，使醱酵產物量提升。
本研究利用基因重組技術，成功地將厭氧菌 Clostridium phytofermentans 中的木糖異構酶 XI 基因建構於載體 pET-14b，並將此質體轉殖於 E.coli BL21(DE3) 中進行木糖異構酶的表現生產。針對 pET14b-CpXI 所產異構酶進行活性測試，其最佳反應條件為 pH 7.5、45 ℃，比活性為 1.85 U/mg，此外，與商業化之 Streptomyces 來源之固定化木糖異構酶相較，其木酮糖平衡產率約低 10 %。在以木質醋酸菌 Acetobacter xylinum BCRC 12334 與剔除葡萄糖脫氫酶 (GDH) 基因之突變菌株生產細菌纖維素時，發現木糖不易代謝，且纖維素產量亦低於木酮糖為碳源，細菌纖維素轉化率可達 11.26 %，木酮糖有利於突變菌株醱酵生產纖維素。以同步異構化醱酵培養時，木糖含量能有效地被轉換成木酮糖而代謝；細菌纖維素產量可達 0.8
g/ L，與木糖相較，產量可提高四倍。對麵包酵母之酒精醱酵，可將 12.81 g/L 之木酮糖轉化產生出 1.01 g/L 之酒精。

Xylose and glucose are the most abundant fermentable sugars that can be obtained from lignocellulosic biomass. However, xylose is not easily metabolized by S.cerevisiae and A. xylinum to produce bacterial cellulose and ethanol. In this present study, D-xylose was converted to its keto isomer, D-xylulose, by recombinant xylose isomerase and immobilized glucose isomerase, sweetzyme. Xylulose can be metabolized by S.cerevisiae and A. xylinum. Simultaneous isomerization and fermentation (SIF) can shift the isomerization equilibrium in favor of xylulose generation and benefits theproduction of fermentation products.
The gene of xylose isomerase from an anaerobic bacterium Clostridium phytofermentans, was cloned and successfully expressed in E.coli BL21 (DE3) using pET14b as a vector. The xylose isomerase showed maximum activity of 1.85U/mg at pH 7.5 and 45℃. Besides, compared with immobilized glucose isomerase, equilibrium concentration of xylulose was about 10% lower. Bacterial cellulose produced in D-xylose medium was much less than the amount of xylose consumed during static cultivation of A. xylinum. The A. xylinum GDH mutant was a better cellulose producer than when in D-xylose/D-xylulose mixed medium was employed. The sugar to cellulose conversion yield of 11.26 % can be reached. D-xylose became fermentable sugar for acetic acid bacterial and yeast if xylose isomerase was added to the medium. The bacterial cellulose productivity of A. xylinum mutant strain with XI addition was about 4 times higher than that host strain. When S.cerevisiae, was employed to utilize xylose with the help of xylose isomerase, approximately 1.01 g/L ethanol could be produced from 12.81 g/L xylose.

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