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研究生:任耕毅
研究生(外文):JEN, KENG-YI
論文名稱:環境因子對本土菌株所生產之角蛋白酶活性之影響
論文名稱(外文):Effect of Environmental Factors on Keratinase Activity of Native Strain
指導教授:吳建一
指導教授(外文):WU, JANE-YII
學位類別:碩士
校院名稱:大葉大學
系所名稱:生物產業科技學系碩士在職專班
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:54
中文關鍵詞: Bacillus sp. 角蛋白酶 飼料添加劑 羽毛
外文關鍵詞:Bacillus sp.KeratinaseFeed additionFeather.
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自本地養雞場附近土壤中所篩選出具角蛋白分解能力之菌株,經初步鑑定為Bacillus菌屬之thuringiensis,初步命名為B. thuringiensis Wu2,可在兩週內將羽翼及部分羽軸完全分解。以傅立葉轉換紅外光譜儀(FTIR)的分析,經B. thuringiensis Wu2降解後,羽毛結構官能基之間的鍵結均遭角蛋白酶破壞而減少。B. thuringiensis Wu2菌酛培養於37℃及pH 7的環境下,在第68小時其菌量可達到最高1.135 g/L。
B. thuringiensis Wu2培養於pH 7,角蛋白酶之最佳活性表現溫度為40℃,經36小時培養角蛋白酶活性可達3.50 kU/mL。當培養環境溫度55℃以上、pH值低於3或高於11,培養72小時後所測得角蛋白酶的活性趨近於0,表示在強酸、強鹼及高溫的環境下B. thuringiensis Wu2耐受性低或在此環境下分泌的酵素的能力下降。培養基中額外添加不同來源的氮原素(Peptone、Urea、NH4Cl及NaNO3)對角蛋白酶的活性並無助益,甚至有導致角蛋白酶活性下降的現象。
The bacteria Bacillus thuringiensis Wu2 was isolated from poultry farm soil, it can degraded the feather completely in two weeks. The degraded feather was observed with Fourier transform-infrared (FTIR), the bonds between the function group of feather were discredited and decrease. Under the culture condition 37℃ and pH 7, the biomass were reached the maximum 1.135 g/L at 68 hour. The maximum activity was 3.50 kU/mL and the optimal pH and temperature of keratinase activity of B. thuringiensis Wu2 was pH 7 and 40℃, respectively otherwise addition the nitrogen source of Peptone、Urea、NH4Cl and NaNO3 into the medium,it doesn’t significance to the keratinase activity.
目錄

封面內頁 頁次
簽名頁
授權書---------------------------------------------------------------------------iii
中文摘要---------------------------------------------------------------- -------iv
英文摘要------------------------------------------------------------------------v
誌謝------------------------------------------------------------------------------vi
目錄------------------------------------------------------------------------------vii
圖目錄---------------------------------------------------------------------------x
表目錄---------------------------------------------------------------------------xi

1. 前言---------------------------------------------------------------------1
2. 文獻回顧---------------------------------------------------------------3
2.1蛋白酶(protease)之簡介與工業上的應用---------------------3
2.1.1蛋白酶的簡介----------------------------------------------------3
2.1.2蛋白酶在工業上的應用----------------------------------------3
2.2角蛋白酶(keratinase)的介紹------------------------------------4
2.2.1角蛋白酶之簡介-------------------------------------------------4
2.2.2 角蛋白酶之特性------------------------------------------------6
2.2.3角蛋白酶之生產菌株-------------------------------------------6
2.3角蛋白(keratin)之簡介----------------------------------------------9
2.3.1角蛋白的來源------------------------------------------------9
2.3.2角蛋白的結構----------------------------------------------------10
2.3.3角蛋白的特性----------------------------------------------------14
2.4角蛋白酶生產菌株所生產之角蛋白酶其應用與發展----------14
2.4.1 飼料工業之應用------------------------------------------------14
2.4.2 皮革工業---------------------------------------------------------18
2.4.3 清潔劑工業------------------------------------------------------18
2.4.4 醫藥及美容用品------------------------------------------------19
3. 材料與方法-----------------------------------------------------------------20
3.1 藥品及儀器設備------------------------------------------------------20
3.1.1藥品---------------------------------------------------------------20
3.1.2儀器設備---------------------------------------------------------21
3.2 菌株來源--------------------------------------------------------------22
3.3 培養基----------------------------------------------------------------23
3.4 培養方法--------------------------------------------------------------23
3.4.1不同pH培養基之培養方法-----------------------------------23
3.4.2不同溫度之培養方法----------------------------------------23
3.4.3不同氮源培養基之培養方法-------------------------------24
3.5分析方法-----------------------------------------------------------24
3.5.1 角蛋白酶活性之分析方法--------------------------------24
3.5.2 蛋白質定量之分析方法(Folin-phenol法)--------------26
3.5.3掃描式電子顯微鏡(SEM)檢測方法--------------27
3.5.4氨基酸組成之分析------------------------------------------27
3.5.5 傅立葉轉換紅外線光譜分析--------------------------------28
4. 結果與討論----------------------------------------------------------29
4.1 角蛋白酶生產菌株之篩選----------------------------------------29
4.2 不同培養溫度對酵素活性的影響---------------------------31
4.3 不同pH值的環境對角蛋白酶活性的影響-----------------------32
4.4 不同氮源的環境對角蛋白酶活性的影響------------------------33
4.5 雞羽毛經B. thuringiensis Wu2粗酵素作用後之胺基酸組成
與官能基變化--------------------------------------------------------34
5. 結論-------------------------------------------------------------------46
參考文獻------------------------------------------------------------------48
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