利用高酯化酒精不溶物管柱去除商业果胶酵素中果胶甲基酯酶以降低.docVIP

利用高酯化酒精不溶物管柱去除商業果膠酵素中果膠甲基酯酶以降低柳丁酒中甲醇含量之探討Reduction the methanol content of sweet orange wine with high methyl-esteration cross linking alcohol insoluble substance to depectinmethylesterase pectinase 研究生：吳嘉峯 Wu, Gia-Feng 指導教授：江啟銘 Jiang, Chii-Ming 吳明昌 Wu, Ming-Chang 商業果膠酵素在製酒過程中扮演重要角色，可促進色澤、芳香物質之萃取、過濾、澄清及增加產量，因此商業果膠酵素應用，在製酒上是不可缺少的加工步驟。但商業果膠酵素中果膠酯酶會水解果膠碳六上甲基酯鍵，產生游離羧基和甲醇，其中甲醇對人體會造成頭痛、暈眩、嘔吐、視力模糊，嚴重甚至會造成失明、死亡，因此將果膠酯酶分離去除頗為重要。以自製之80酯化度 80 DE 、50酯化度 50 DE 之CL-AIS管柱，經酯化度測定後其酯化度分別為80DE與50 DE，將市售商業果膠酵素通入管柱內，經0-0.1 M NaCl沖提可於DE 為80之管柱可將PE自市售商業果膠酵素中分離且PL之純化倍率為6.7倍，回收率則高達8.3%，顯示以調整CL-AIS親和性膠體之酯化度可快速將商業果膠酵素中PE與PG、PL分離，並獲得PL PL：450 unit/mg、PG：8.3 unit/mg 、純化倍率6.7、回收率84.3% 與PE PE：100 unit/mg、PG：6.5 unit/mg、PL 82.4 unit/mg、純化倍率4.8、回收率68% 。進一步將其應用於柳丁酒釀造上，探討去除PE之商業果膠酵素其在水果酒釀造中降低甲醇生成之可行性，結果發現柳丁酒在發酵過程中澄清度隨著時間的增加而增加，其原因為果膠酵素作用所致，使得果汁中的果膠被分解而使得透光度提高，凡添加酵素組別其透光度均顯著性比控制組高。而在甲醇生成量上，結果顯示添加商業果膠酵素之柳丁酒其甲醇含量約60 ppm，而單獨添加PE的柳丁酒則高達85 ppm，皆顯著高於其他各組，而在其他酒類物理特性部，則證實將PE單獨去除並不會對於酒類其他物理特性造成顯著的影響。 果膠分解酵素、酯化CL-AIS、甲醇ectinolytic enzymes are usually used in winemaking for clarification and yield increasing of juices and wine making , extraction of oil and color from vegetables and fruits , so commercial pectinolytic enzyme is important in making wine. The PE can catalysis the de-esterification reaction, the methyl ester group on the C6 carboxyl of D- galacturonic acid in the pectin could be hydrolyzed by PE to produce the free carboxyl group and the methanol. Methanol usually causes headache, vertigo, vomiting, blurred vision, blindness and even death, so, the separation of pectinesterase from pectinase is needed in wine making. The commercial pectinolytic enzymes is affinity to the 83 DE and 50 DE cross linking alcohol insoluble substances, the PL of pectinase could be separated with the PE, the remaining PL containing 445 unit/mg of PG and 8.3 unit/mg of PL, the purified fold of PL was 6.7 and the recovery was 84.3%, the separated PE containing 100 unit/mg PE, PG 6.5 unit/mg and PL 82.4 unit/