纤维素分解酵素生产菌的筛选培养条件及糖化作用之探讨.DOC

纖維素分解酵素生產菌的篩選培養條件及糖化作用之探討 Research on Screening and Incubation Conditions of Cellulolytic Microorganisms and their Sacchariation on Cellulose Materials 研究生：黃妃君 Hun, Fa-Chung 指導教授：鄧德豐 【摘要】 用簡單培養方法從大自然界分離具有纖維素分解酵素生產菌.得到三株具有較高的纖維分解酵素活性的菌株分別命名為GG12,GG14,GG24三株菌株經過培養和顯微鏡上的型態觀察,初步鑑定GG12和GG24為Aspergillus 菌屬,而GG14為Penicillum菌屬.這三株菌株經過培養來尋找出最適合纖維素分解酵素的生產條件.最適合纖維素分解酵素的生成之碳源是羧甲基纖維素,羧甲基纖維素和NH4NO3配製最好生成纖維素分解酵素之碳氮比值是10:1,同時發現這三株菌株培養於30℃ pH5.0和振盪轉速為130rpm可以得到較高纖維素分解酵素活性.三株菌株所形成之纖維素分解酵素最適合反應pH值是PH5.0-6.0,但是GG24菌株的CMCase及B-glucosidase比GG12和GG14的CMCase及B-glucosidase更具耐酸性.這三株菌株所形成的酵素最適合反應溫度是60℃,它們再70℃加熱10分鐘仍十分安定.GG24菌株的CMCase活性比GG12和GG14菌株耐高溫,而GG12和GG14的B-glucosidase反比GG24的耐高溫,因為這三株菌株所形成的纖維素分解酵素均耐高溫允許在高溫下進行糖化作用.在實用上,酵素在進行糖化作用時更為安定.使用0.1M檸檬酸緩衝液配製纖維素培養基可使纖維素分解酵素活性增加.本篇研究也使用混合培養來增加纖維素分解酵素的形成.混合培養採取相化(phasing)和孢子比兩種方式進行196小時之培養.在GG12/GG24的相化培養中,延後12小時加入有較佳酵素活性.在GG14的相化培養中,延後8小時加入有較佳酵素活性.在GG12:GG24的不同孢子比實驗中發現GG12:GG24孢子比在1:1和1:2之間有較佳纖維素分解酵素活性.纖維素分解酵素的糖化能力受到酵素之成份及纖維素得結構影響巨大.使用四種酵素系統如GG12酵素,GG24酵素,GG12和GG24混合培養酵素及GG12/GG24相化酵素作用於不同纖維素物質來比較糖化能力,以GG12和GG24相化的酵素作用於牧草可得到75%之最大糖化轉換率. 關鍵字：纖維素 糖化作用 相化 【Abstract】 The cellulolytic fungi was screened from the nature by the simple culturaltest. Three strains of higher activity of cellulolytic fungi were isolated,named as GG12,GG14 and GG24. These three strains of fungi were identified bytheir morphological examination on different agar medium and with microscope.GG12 and GG24 were identified as the genera of Aspergillus sp.and GG14 was asthe genera of Penicillium sp.these three strains of fungi were incubated from searching the optimum conditions of cellulolytic enzyme . the carboxymethyl cellulose was the best carbon source for these strains to produce cellulolytic enzymes.the best C/Nratio of CMC and NH4NO3 in the medium for producing cellulolytic enzymes was 10:1 . we also found these three strains three strains of fungi having highercellulolytic enzymes when cultivated at 30℃,pH=5.0 with cellulolytic enzyme when cultivated at 30℃,pH5.0, shaking 1