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研究生:林明義
論文名稱:利用酵母菌生產生物乳化劑之研究
論文名稱(外文):Study on the production of bioemulsifier by yeast
指導教授:古源光
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:食品科學系
學門:農業科學學門
學類:食品科學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:106
中文關鍵詞:Pichia anomala生物乳化劑培養條件
相關次數:
  • 被引用被引用:6
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本研究由自然界中腐敗之柚子、梅子、桃子、葡萄及水蜜桃等水果,篩選分離菌株,篩得菌株共有403株,進行表面張力及乳化能力之試驗,挑選出能生產生物乳化劑,且對煤油具乳化活性,選定NPUST-D26為本研究之試驗菌株。NPUST-D26菌株經食品工業發展研究所菌種保存中心鑑定為Pichia anomala。
針對P. anomala NPUST-D26菌株最適培養基之探討,此菌株對於不同碳源(可溶性醣類如葡萄糖,蔗糖,果糖及麥芽糖與非水溶性蔬菜油之沙拉油,玉米胚芽油,葵花油,花生油及橄欖油等)皆生長良好。氮源利用性方面,對於各種氮源(無機氮源之銨鹽如CH3COONH4、NH4Cl、(NH4)2SO4、NH4NO3、(NH4)2HPO4與硝酸鹽如KNO3, NaNO3;有機氮源如尿素、Peptone、BATCOTM peptone、Tryptone peptone等)皆生長良好。進而選定蔗糖與醋酸銨為其碳源及氮源,並探討其濃度,還包括無機鹽類(磷酸鹽與硫酸鎂等)及酵母抽出物之濃度,結果顯示,P. anomala NPUST-D26菌株之最適培養基組成為:每公升培養基含Sucrose 1 % (wt)、CH3COONH4 0.3 % (wt)、K2HPO4 0.01 % (wt)、Yeast extract 0.01 % (wt)、MgSO4‧7H2O 0.002 % (wt)、pH 5.5±0.2。
P. anomala NPUST-D26菌株之生物乳化劑為與細胞壁結合或細胞內之形式,故以熱萃取方式,利用高壓殺菌釜處理120 min,萃取生物乳化劑,每公升培養液可產生1.85 g之粗生物乳化劑。其臨界微胞濃度(CMC)為0.3~0.5 %,可將蒸餾水之表面張力由72 dyne/cm降至53 dyne/cm。其組成包括粗蛋白34.9 %、粗脂肪0.34 %、水分4.4 %、灰分36.8 %及還原醣含量15 %,推測可能為醣蛋白之生物乳化劑,但有待確定其離子性。0.5%之粗生物乳化劑,在0~5%之氯化鈉濃度、酸鹼值2~10及溫度6~100℃之間,對煤油之乳化能力為72±1.5 %左右,且可將水之表面張力降至54±1.2 dyne/cm左右。以5公升小型醱酵槽擴大培養之操作條件為:培養溫度25℃,通氣量0.5 vvm,攪拌速率400 rpm。每公升培養液可產生2.18克之粗生物乳化劑。

目錄
頁次
中文摘要 I
Abstract III
目錄 V
圖目錄 VIII
表目錄 XI
壹、緒言 1
一、研究動機 1
二、研究目的 3
貳、文獻回顧 5
一、乳化現象 5
(一)、HLB值(Hydrophile-Lipophile Balance) 6
(二)、臨界微胞濃度(Critical Micelle Concentration, CMC) 8
二、食品乳化劑/安定劑 11
(一)、乳化劑 12
(二)、安定劑 16
三、生物乳化劑 17
(一)、醣脂類(glycolipids) 20
(二)、脂胜肽(lipopeptides) 24
(三)、脂肪酸與磷酸類 (Fatty acids and phospholipids) 27
(四)、聚多醣─蛋白質乳化劑Bioemulsans (polysaccharide- protein surfactants ) 31
(五)、其他高分子生物乳化劑(Other high-molecular-mass bioemulsifiers) 36
四、生物乳化劑的產生可能的原因及所面臨的問題 39
參、材料與方法 42
一、試驗菌株 42
二、培養基組成 42
三、生物乳化劑生產菌株之分離及篩選 44
四、菌學鑑定 45
五、最適培養基之探討 46
六、小型醱酵槽中生產條件之探討 47
七、粗生物乳化劑 47
八、生物乳化劑之物理特性探討 47
九、臨界微胞濃度(Critical Micelle Concentration, CMC) 48
十、分析方法 48
十一、實驗設備 52
十二、藥品 53
肆、結果與討論 54
一、生物乳化劑生產菌株之篩選與菌種鑑定 54
二、最適培養基組成份之探討 58
1. 添加煤油、葡萄糖與沙拉油對P. anomala NPUST-D26生長情形及乳化指數之影響 58
2. 不同碳源培養基對P. anomala NPUST-D26生長情形及乳化指數之影響 62
(1)水溶性之醣類對生長情形及乳化指數之影響 62
(2)非水溶性之蔬菜油對生長情形及乳化指數之影響 65
3. 不同氮源培養基對P. anomala NPUST-D26生長情形及乳化指數之影響 69
(1) 無機氮源對P. anomala NPUST-D26生長情形及乳化指數之影響 69
(2) 有機氮源對P. anomala NPUST-D26生長情形及乳化指數之影響 71
4. 培養基內其它成份添加對P. anomala NPUST-D26生長情形及乳化指數之影響 71
三、粗生物乳化劑之分離回收方法之探討 73
四、生物乳化劑之特性探討 82
1. 不同氯化鈉濃度、酸鹼值及溫度對乳化指數與表面張力之影響 82
2. 臨界微胞濃度(Critical Micelle Concentration, CMC) 86
3. 生物乳化劑離子性之分類 86
五、生物乳化劑之組成分析 88
六、小型發酵槽操作條件之探討 91
伍、結論與建議 93
陸、參考文獻 95
圖目錄
頁次
圖2-1 乳化劑的HLB值對其性狀、用途的影響 7
圖2-2 乳化劑濃度與各種物理化學性質之關係 9
圖2-3 乳化劑溶於水中的情形 10
圖2-4 化學合成食品乳化劑分子之可能的結構 13
圖2-5 醣脂之結構 21
圖2-6 脂胜肽之結構 25
圖2-7 corynomycolic acid之結構 28
圖2-8 磷脂之結構 30
圖2-9 Emulsan之結構 32
圖2-10 Emulsan可能之型態 34
圖2-11 A. calcoaceticusMM5細胞黏附於乳化液滴之顯微照相 35
圖2-12 Mannoprotein於細胞壁的構成及其結構 38
圖4-1 Pichia anomala之顯微照相 55
圖4-2 不同碳源對P. anomala NPUST-D26生長情形之影響 59
圖4-3 不同碳源對P. anomala NPUST-D26乳化指數之影響 60
圖4-4 不同醣類對P. anomala NPUST-D26生長情形之影響 63
圖4-5 不同醣類對P. anomala NPUST-D26乳化指數之影響 64
圖4-6 不同蔬菜油對P. anomala NPUST-D26生長情形之影響 66
圖4-7 不同蔬菜油對P. anomala NPUST-D26乳化指數之影響 67
圖4-8 不同蔗糖濃度對P. anomala NPUST-D26生長情形與乳化指數之影響 68
圖4-9 不同氮源對P. anomala NPUST-D26生長情形與乳化指數之影響 70
圖4-10 不同醋酸銨濃度對P. anomala NPUST-D26生長情形與乳化指數之影響 72
圖4-11 微量元素對P. anomala NPUST-D26生長情形與乳化指數之影響 74
圖4-12 不同硫酸鎂濃度對P. anomala NPUST-D26生長情形與乳化指數之影響 75
圖4-13 不同磷酸氫二鉀濃度對P. anomala NPUST-D26生長情形與乳化指數之影響 76
圖4-14 不同酵母抽出物濃度對P. anomala NPUST-D26生長情形與乳化指數之影響 77
圖4-15 不同處理時間對粗生物乳化劑產量與乳化指數之影響 80
圖4-16 不同濃度粗生物乳化劑對煤油及沙拉油乳化指數之影響 81
圖4-17 氯化鈉濃度對0.5%生物乳化劑乳化指數與表面張力之影響 83
圖4-18 不同酸鹼值對0.5%生物乳化劑乳化指數與表面張力之影響 84
圖4-19 不同溫度對0.5%生物乳化劑乳化指數與表面張力之影響 85
圖4-20 粗生物乳化劑之臨界微胞濃度 87
圖4-21 Epton法之結果 89
圖4-22 碘化鉀-碘法之結果 90
圖4-23 P. anomala NPUST-D26置於5公升醱酵槽與500毫升Hinton氏三角瓶以不同操作條件培養之影響 92
表目錄
頁次
表2-1 一般公認安全性之乳化劑 14
表2-2可作為食品添加物之乳化劑 15
表2-3各種微生物生產之界面性活劑 19
表2-4 鼠李醣脂之產品 23
表2-5 生產surfactin相關分子之醱酵操作條件及產率 26
表2-6 微生物生產生物界面活性劑之可能方式 40
表3-1 基礎培養基:馴養及篩選用 42
表3-2 YM斜面培養基:菌株保存用 43
表3-3 YM培養基:菌株活化用 43
表3-4 最適培養基:擴大培養用 44
表4-1 P. anomala NPUST-D26的生理學特徵 56
表4-2 從數種酵母菌中熱萃取之乳化劑 78
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