醱酵生產日本麴菌、黑麴菌、黃麴菌和出芽短梗黴的果糖轉移酶

Fermentative Production of Fructosyltransferase from Aspergillus japonicus, A. niger, A. oryzae and Aureobasidium sp.

曾柏瑞(Bo-Ruei Tseng)

指導教授 ： 許垤棋

大同大學/工程學院/生物工程學系所/碩士(2010年)

摘要

果糖轉移酶可催化蔗糖生成果寡糖，果寡糖是一種重要的益生源。工業上使用的果糖轉移酶主要來自黴菌，本研究利用五公升醱酵槽分別培養四株黴菌，包括：日本麴菌（Aspergillus japonicus）、黑麴菌（A. niger）、黃麴菌（A. oryzae）以及出芽短梗黴（Aureobasidium sp.），以2 %酵母萃取物為氮源，不同濃度的蔗糖為主要的碳源，利用批次醱酵和饋料醱酵生產果糖轉移酶。日本麴菌的果糖轉移酶是胞內酵素，黑麴菌和黃麴菌是胞外酵素，而出芽短梗黴則是胞內胞外都有。饋入蔗糖可提高日本麴菌、黑麴菌、黃麴菌和出芽短梗黴的酵素產量，顯示四株菌的果糖轉移酶具有可誘導性，調整蔗糖的濃度或饋入速率的快慢可提高果糖轉移酶的產量。以蔗糖為基質進行饋料批次醱酵，起始的醱酵體積是二公升，蔗糖體積是一公升，當75% (w/v)蔗糖溶液的饋入速度從55.6 mL/h降至27.8 mL/h，可提高日本麴菌的酵素產量，從606 U/mL至704 U/mL。醱酵過程不只蔗糖是誘導劑，蔗糖生成的果寡糖分子也有誘導的作用。四株菌的酵素產量以日本麴菌最高852 U/mL，其次為黑麴菌613 U/mL、出芽短梗黴262 U/mL，酵素活性最低的是黃麴菌208 U/mL。

關鍵字

日本麴菌 ； 黑麴菌 ； 黃麴菌 ； 出芽短梗黴 ； 醱酵槽 ； 果糖轉移酶 ； 果寡糖

並列摘要

Fructooligosaccharides (FOS) are important prebiotic substances. Commercial production of FOS is carried out through the catalytic reaction of fungal fructosyltransferase (FTase) on sucrose. This work was engaged in production of FTase from Aspergillus japonicus, A. niger, A. oryzae and Aureobasidium sp. using a 5-L fermentor. FTase is intracellular in A. japonicus, but extracellular in A. niger and A. oryzae. However, it is both intracellular and extracellular in Aureobasidium sp. In the fermentation process, the culture medium consisted of 2% yeast extract as nitrogen source and varied amount of sucrose as carbon source. FTase activities produced both in batch fermentation and fed-batch fermentation were compared. During the fermentation with A. japonicus, A. niger, A. oryzae or Aureobasidium sp., FTase activities increased when sucrose is continuously added to the culture broth, indicating FTase of these strains are inducible. Furthermore, production of FTase from A. japonicus, A. niger, A. oryzae and Aureobasidium sp. can be promoted by varying the initial concentration or feeding rate of sucrose added to the culture during fermentation. The fed-batch fermentation was carried out with an initial volume of 2 L and 1 L of sucrose solution was fed gradually into the fermentor. During the fermentation with A. japonicus, when the feeding rate of 75% (w/v) sucrose was decreased from 55.6 mL/h to 27.8 mL/h, the enzyme production is increased from 606 U/mL up to 704 U/mL. In the cases of this work, not only sucrose but also FOS molecules formed during fermentation are inducers for the FTase. The yields of FTase are ranked in the following sequence, from the highest to the lowest, Aspergillus japonicus (852 U/mL), A. niger (613 U/mL), Aureobasidium sp. (262 U/mL), and A. oryzae (208 U/mL).

並列關鍵字

Fermentor ； Aureobasidium ； Aspergillus oryzae ； Aspergillus niger ； Aspergillus japonicus ； Fructosyltransferase ； FOS

參考文獻

黃伯欣。2008。利用日本麴菌進行液態發酵和固態發酵生產果糖轉移酶。大同大學生物工程研究所碩士論文。

Antošová, M., V. Illeová, M. Vandáková, A. Družkovská, and M. Polakovič. 2008. Chromatographic separation and kinetic properties of fructosyltransferase from Aureobasidium pullulans. J. Biotechnol. 135: 58-63.

Duan, K. J., J. S. Chen, and D. C. Sheu. 1994. Kinetic studies and mathematical model for enzymatic production of fructooligosaccharides from sucrose. Enzyme Microb. Technol. 16: 334-339.

Godshall, M. A. .2007. Future directions for the sugar industry. http://www.spriinc.org/buton10bftpp.html

Maiorano, A. E., R. M. Piccoli, E. S. da Silva, and M. F. de Andrade Rodrigues. 2008. Microbial production of fructosyltransferases for synthesis of pre-biotics. Biotechnol. Lett. 30: 1867–1877.