Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/51569
標題: Characterization, Optimization of Xylanase Production from Aspergillus carneus M34 in Solid-State Fermentation and Its Application in Xylooligosaccharides Production.
Aspergillus carneus M34 聚木糖酶特性分析、最適化 固態發酵生產條件與其應用於木寡醣生產之探討
作者: Yang, Chieh-Hsiu
楊杰修
關鍵字: Aspergillus carneus M34
Aspergillus carneus M34
聚木糖酶
固態發酵
木寡醣
Xylanase
Solid-state fermentation
Xylooligosaccharides
出版社: 食品暨應用生物科技學系所
摘要: Xylanase production strain, Aspergillus carneus M34, which was isolated from soil by our laboratory, was used to study the growth condition for optimal xylanase production, extraction methods and enzyme characterization by solid state fermentation used coba husk and corn steep liquor for the medium. In addition, possibility of the crude enzyme degrading coba husk hemicellulose to produce xylooligosaccharides was evaluated. Solid-state was the better fermentation type to ferment coba husk for xylanase production of Aspergillus carneus M34. In one factor test, the better cultivation conditions as follows: inoculums size 106 spores g-wm-1, cultivation temperature 25℃, initial pH value of medium pH 8.0. For optimization of the cultivation conditions, central composite design and response surface methodology were practiced. The optimal cultivation conditions were 25.6℃ of cultivation temperature, initial pH value of medium pH 7.96, the predicted maximum xylanase activity under optimal conditions was 6,781 U gdm-1. This represented 3.9-fold gain in xylanase activity. The better extraction conditions as follows: extraction temperature 25℃, extraction time 1 hour, pH of extractive pH 5.0 ~ 6.0, Tween 80 concentration of extraction solution 0.1%. The test of crude enzyme degradation of coba husk hemicellulose. The optimal pH and temperature were pH 5.0 and 50 ~ 60℃, respectively. It also exhibited a good pH stability at range of pH 4.0 to 7.0. In respect to thermostability, the residual activity of xylanase activity was 80% at 50℃ for 70 min. The crude enzyme hydrolyzed product contented xylooligosaccharides. The predicted xylooligosaccharides yield pre kg coba husk were xylobiose 61 g, xylotriose 11 g and xylotetraose 22 g under 30 min reaction.
本研究乃利用本實驗室自土壤中篩選出具有聚木糖酶活性之Aspergillus carneus M34,以茭白筍殼與玉米浸漬液作為固態發酵主要基質,探討生產聚木糖酶之較適條件,並找出較適粗酵素液之萃取條件,進而探討酵素特性。此外,本研究利用此粗酵素液降解茭白筍殼半纖維素,以評估生產木寡醣之可能性。 實驗結果顯示,以A. carneus M34發酵茭白筍殼以固態發酵進行效果較佳。單一因子試驗中,較適生產聚木糖酶之培養條件如下:接種量為106 spores g-wm-1,培養溫度為25℃,培養基起始pH值為pH 8.0。藉由中心混成設計及反應曲面法之探討後,最適化培養條件為培養溫度25.6℃,培養基起始pH值為pH 7.96,預估聚木糖酶活性可達 6,781 U g-dm-1,較未最適化培養條件所能產生 1,721 U g-dm-1,可增加 3.9 倍。萃取粗酵素液之較適萃取條件如下:萃取溫度 25℃、萃取時間 1 小時、萃取溶液 pH 值為 pH 5.0 ~ 6.0、萃取溶液 Tween 80濃度 0.1%。 於利用本實驗生產之聚木糖酶降解茭白筍殼半纖維素試驗中,其最適作用 pH 值為 pH 5.0、最適作用溫度為 50 ~ 60℃,酵素在 pH 4.0~6.0 穩定性較佳,且於 50℃下作用 70 分鐘分鐘後,仍有 80% 以上之聚木糖酶活性。分析其水解產物中,含有木二糖、木三糖及木四糖等寡醣。預估每公斤之茭白筍殼所能產生之木寡醣量為(酵素作用時間 30 min):木二糖 61 g;木三糖 11 g;木四糖 22 g。
URI: http://hdl.handle.net/11455/51569
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