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  • 學位論文

溫度對甘藷β-澱粉酶活性影響之研究

Effect of temperature on β-amylase activity in sweet potato

指導教授 : 李平篤

摘要


β-澱粉酶(β-amylase, EC 3.2.1.2,簡稱BA) 催化水解澱粉鏈之非還原端 α-1,4-糖甙鍵,釋出β-maltose而留下β-limit dextrin,分類上屬於外切澱粉酶。 本論文針對不同品種之甘藷測定BA活性,選擇總活性最高之60℃樣本與對照組進行純化,測定生化性質加以比較,來探討溫度對BA活性之影響。純化將粗抽液經硫酸銨100%沈澱後,進行製備式電泳可得到相當純的BA。 對照組與60℃樣本對基質澱粉之 Km 值分別為 6.02 與 2.95%。酵素最適反應溫度均為60℃,對照組最適反應pH為 5,60℃樣本為 4.5。對照組與 60℃樣本之活化能分別為 3.35 與 4.2 kcal/mol;酵素活性會受到汞、銅、銀離子的抑制,除了汞離子外,60℃樣本對金屬離子的耐受性均較對照組差。化學修飾劑β-mercaptoethanol、DTT、PMSF中除了DTT外,60℃樣本受到的影響較對照組來得大。

關鍵字

溫度 甘藷 β-澱粉酶

並列摘要


β-amylase (EC 3.2.1.2) is an exoamylase that releases successive maltose units from the nonreducing end of a polysaccharide chain by hydrolyzing of α-1,4-glucan linkages. In this article, BA activities were measured in different varieties of sweet potato, and chose the highest total activity sample under 60℃ treatment and control sample to purify. And put to compare their biochemical characteristics and the effects of temperature on BA activity. Crude extraction precipitated by ammonium sulfate 100% then run preparative electrophoresis can get pure enzyme. Km values for starch substrate of control sample and 60℃ sample were estimated to be 6.02 and 2.95%, respectively. Optimal temperature for both samples are 60℃. Optimal pH value for control sample is 5, and for 60℃ sample is 4.5. Activation energy for control sample and 60℃ sample is 3.35 and 4.2 kcal/mol, respectively. BA activity could be inhibited by Hg2+, Cu2+ and Ag+. Except for Hg2+, tolerance to metal ions of 60℃ sample is less than control sample. Chemical modified reagents, including β-mercaptoethanol, DTT, PMSF also influence BA activity. Except for DTT, influence of chemical modified reagents of 60℃ sample is more than control sample.

並列關鍵字

temperature sweet potato β-amylase

參考文獻


Fan ML (1975) Partial purification and properties of potato beta-amylase. Taiwania 20: 132-138
Ann YG, Iizuka M, Minamura N (1989) Evidence for existence of an active monomer of sweet potato β-amylase. Agric. Biol. Chem. 53(11): 3109-3110
Arruda EJ, Santana CC (2003) Phenylboronate-chitosan resins for adsorption of beta-amylase from soybean extracts. Appl Biochem Biotechnol 105 -108: 829-42
Baba T, Kainuma K (1987) Partial hydrolysis of sweet potato starch with β-amylase. Agric. Biol. Chem. 519(5): 1365-1371
Beck E, Ziegler P (1989) Biosynthesis and degradation of starch in higher plants. Annu Rev Plant Physiol Plant Mo1 Biol 40: 95-117

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