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題 名 | 北蕉黑星病葉酪胺酸酶同功酶之生化特性=The Biochemical Properties of Tyrosinase Isozymes from the Banana Leaf with Black Spot Disease |
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作 者 | 朱惠鈴; 吳鴻程; 郭建民; 葉東柏; 蕭介夫; | 書刊名 | 中國農業化學會誌 |
卷 期 | 37:1 1999.03[民88.03] |
頁 次 | 頁95-104 |
分類號 | 435.371 |
關鍵詞 | 北蕉葉; 黑星病; 酪胺酸酶; 同功酶; 生化特性; Banana leaf; Black spot disease; Tyrosinase; Isozyme; Biochemical property; |
語 文 | 中文(Chinese) |
中文摘要 | 酪胺酸�t由北蕉黑星病中抽出,經過DEAE-Sepharose CL-6B管分離後得 到5個具酪胺酸�t活性部份,F5∼F5。以Superose 6 管柱層析測得F1∼F5分子量 分別為135KD,75KD,50KD,120KD,及100KD。最適溫度F1及F4為40℃, F2、F3及F5為35℃。最適pH值F1及F3為6.8,F2及F5為7.5,F4為8.5或以 上。銅離子於高濃度(>0.6mM)時會抑制F1∼F5,而低濃度(<0.3mM)時會活F1 ∼F5酵素活性所有同功�t酵素活性幾乎完全被氰化鈉抑制,推測五種同功�t都為含 金屬酵素,其活性中心可能含銅離子。黑星病葉中酪胺酸�t同功�tF1與同品種健康葉 純化之酪胺酸�t於分子量,最適pH值、最適溫度金屬離子及有機化合物之活化或抑 制情形等生化特性均很相似,推測兩者為同一酵素。而黑星病葉之粗酵素抽出液對L- 酪胺酸具有不尋常的高活性,推測此酪胺酸�t對於受黑星病感染之北蕉單酚代謝程序 應有其重要性。 |
英文摘要 | Five isozymes (F1∼F5) of tyrosinase were partially purified from the banana leaf (Musa sapientum cv, Giant Cavendishii, AAA) with black spot disease using DEAE-Sepharose CL-6B separation, The molecular mass of F1 ∼ F5 was determined by means of Superose 6 chromatography o be 135KD,75KD,50KD,120 KD and 100 KD, respectively. The optimum temperature of F1 and F4 was 40 ℃ when L-DOPA was used as the substrate; that of F2,F3 and F5 was 35 ℃. The optimum pH of F1 and F3 was 6.8; that of F2 and F5 was 7.5; AND THAT OF f4 WAS 8.5. Cu ���� inhibited the activity of tyrosinase isozymes of F1 ∼ F5 at concentrations higher than 0.6 mM while it activated enzyme at concentrations lower than 0.3mM. Moreover, the tyrosinase activity activity of F1 ∼ F5 was inhibited completely by sodium cyanide at a concentration of 5mM. It is speculated that the tyrosinase isozymes of F1 ∼ F5 were metalloenzyme, and that Cu ���� was probably required for their active sites. The biochemical properties of the isozyme F1 from the banana leaf with black spot disease, including optimum pH and temperature, molecular mass, and the effects of metal and organic compounds on tyrosinase activity, were similar to that from the healthy leaf, implying that those two were probably the same enzyme. The tyrosinase of the banana leaf with black spot disease exhibited unusually high activity toward L-tyrosine, which suggested that the tyrosinase might be very important for the monophenol metabolism in the banana leaf infected with black spot disease. |
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