几丁质及其衍生物抗菌性之探讨.doc

國立宜蘭大學食品科學系碩士班專題討論題目： 國立宜蘭大學食品科學系碩士班專題討論 題目：以酵素法產製之幾丁寡醣其特性及抗菌活性之研究 The study of antibacterial activity and characterization of enzymatically-prepared chito-oilgosaccharide. 研究生專題討論書面摘要 指導教授：林世斌 老師 報告者：林怡君 報告時間：200 摘要 幾丁寡醣已被證實具有抗菌之效果，目前產製幾丁寡醣的方法分為化學酸水解及酵素水解法。酵素水解較溫和且專一性較高，故為目前產製幾丁寡醣的主要方法。本研究以木黴菌（Trichoderma harzianum）誘發之幾丁質酵素與市售纖維酵酵素及溶菌酶水解幾丁聚醣，發現水解三小時，去乙醯度80％之幾丁聚醣（DD80）黏度會由403.8cp下降至40cp以下，而去乙醯度92％之幾丁聚醣（DD92）其黏度則由367.2cp下降至55cp以下。而以CCHlys其DNS變化少，CCHcel在四至五天時DNS趨於平緩，六天之後DNS又有上升之情形。CCHtri之DNS則是隨著水解天數而有逐漸上升之趨勢。未水解之幾丁聚醣於中性下溶解度差且分子量較大，對於抑制S. aureus之效果（400-800ppm）大於水解物（＞800ppm）。而對於E. coli則是部分水解產物之抑菌效果（50ppm）略高於未水解者（100ppm）。而分子量較小的cellulase水解產物需200ppm才具有抑制E. coli的效果，但DD92-CCHtri及DD80-CCHtri產物只需50ppm即可抑制E. coli。而lysozyme水解產物，其抑菌以DD80-CCHlys較佳，只需50ppm即可抑菌。未來將以HPLC及SEM觀察水解物粒徑大小，進一步確認抑菌有效分子片段之分佈，並瞭解其抑制機制。 關鍵字：幾丁寡醣；幾丁聚醣；幾丁質酵素；溶菌酶；纖維酵素；去乙醯度；抗菌活性；最低抑制濃度 Abstract Chito-oilgosaccharide was demonstrated having antimicrobial activity. The chemical acid and enzymatic hydrolysis are two common methods for producing Chito-oilgosaccharide. Because enzymatic hydrolysis method is milder, and more specificity, and with lower contamin ation, it has becomes a more favorable method then chemical hydrolysis method. In this study, the hydrolysates of two degree of deacetylation with crude chitinase（from Trichoderma harzianum）, commercial cellulase and commercial lysozyme. The results suggested that the hydrolysis for three hours, degree of deacetylation 80% the viscosity will drop to 40cp from 403.8cp, degree of deacetylation 92% the viscosity will drop to 55cp from 367.2cp. The DNS reagent of CCHlys change few, and CCHcel tends towards stability in four to five days, after six days DNS reagent has situation of the rise. CCHtri DNS reagent increased with reaction times. Chtiosan in the neutrality the solubility difference also has high molecular weight, to suppressing S. aureus（400-800ppm）greater than chitosan hydrolyte（＞800ppm）, but to E. coli is chitosan hydrolyte（50ppm）have higher antimicrobia