| Abstract: | 酪胺酸?是黑色素合成的關鍵酵素，藉由酪胺酸?之抑制，可以達到美白功效。本研究將靈芝菌絲體進行液態培養，取上清液、菌絲體與含菌絲體之發酵液，分別以酸(H2SO4)、鹼(NaOH)萃取與酵素分解。選用的酵素包括Neutrase、Celluclast、Lysozyme、Papain及Bromelain。期望可以得到抑制黑色素生成之有效成分。實驗包括：(一)萃取或分解後之上清液抑制DOPA生成效果之比較。(二)不同分解酵素之最佳pH、溫度及作用時間。(三)有效成分之分析等三部分。結果顯示：發酵液經均質後添加Neutrase 、Celluclast 、Papain作用，其上清液抑制DOPA生成效果最好，分別為79.8%、82.7%、88.1%。改變pH、溫度及分解時間，Neutrase 以pH 5.5、30℃、120分鐘作用後，抑制效果提高為93.4%；Celluclast 以pH 5、45℃、240分鐘作用後，抑制效果為95.9%；結果也顯示：使用Neutrase與Celluclast分解時，作用時間越長其抑制效果趨近於100%，而Papain分解時，作用120分鐘之後，其抑制效果逐漸下降。由福林-酚試劑結果顯示，此有效成分可能為含有酚基胺基酸的胜?類。
Tyrosinase is the key enzyme for the synthesis of melanin. The purpose of this study is to try to produce tyrosinase inhibitor obtained from the submerged culture of Ganoderma lucidum Different parts from the fermented product were tested and compared for the effect of inhibitor, which included supernatant, mycelium, and fermentation broth. Inhibiting melanogenes. In additions, extraction or enzyme hydrolysis were also used to improve inhibition effect. The types of enzymes chosen were Neutrase, Celluclast, Lysozyme, Papain, and Bromelain. The experiment could be categorized mainly into three parts. (1)the comparison of various fermented product for bleaching effect (2) the comparison of various enzyme and hydrolysis conditions (pH, temperature and time) for inhibiting melanogenes and bleaching effect. (3) the analysis of effective components. The results show that the polytron treatment of fermentated broth and the addition of Neutrase, Celluclast, and Papain could improve the bleaching effect to 79.8%, 82.7%, and 88.1%. The optimum conditions of enzyme hydrolysis were determined as follows. The treatment of Neutrase at pH5.5, 30℃, and 120 minutes could cause the inhibiting effect to rise to 93.4%. The treatment of Celluclast at pH5, 30℃, and 240 minutes could lead the increase of the inhibiting effect to 95.9%. The results reveal that prolonged hydrolysis of Neutrase and Celluclast could enhance the bleaching effect. However, the prolonged treatment of Papain after 120 min would cause the reverse effect. After the chemical analysis in Folin & Ciocalteus Phenol reagent, the effective component for bleaching was considered to be some types of peptide containing phenolic group. |
