液化澱粉芽孢桿菌 TKU050 所生產之果膠酶的純化與定性

Purification and characterization of a pectinase from Bacillus amyloliquefaciens TKU050

陳建霖(Chien-Lin Chen)

指導教授 ： 王三郎 ； 阮文邦

淡江大學/理學院/化學學系碩士班/碩士(2020年)

https://doi.org/10.6846/TKU.2020.00026

摘要

在這項研究中，使用了來自香蕉（香蕉皮），大米（米糠），橙子（橙皮），咖啡（廢咖啡渣）以及小麥（麥麩）的各種含農業果膠的副產品，提供碳和氮（C / N）養分。通過篩選自淡水土壤經鑑定為液化澱粉芽孢桿菌的這株果膠酶生產菌TKU050，在含0.5% 麥麩中於37℃連續4天表現出最高的果膠酶生產率（0.76 U / mL）。
B. amyloliquefaciens TKU050於較適培養條件發酵所得上清液，經硫酸銨沉澱、High Q column以及HPLC進行果膠酶之純化分離。純化所得TKU050果膠酶之分子量經SDS-PAGE測得為58 kDa，活性回收率與比活性分別為7.24%與0.91U/mg。TKU050果膠酶之最適反應溫度與pH分別為50℃及pH6，熱穩定性及pH安定性分別為<50℃與pH6~9。TKU050果膠酶活性會受Cu2+和SDS所抑制。
用TKU050果膠酶水解得到的產物對乳酸菌的生長具有增強作用。產物具促進乳酸桿菌生長之效果。

關鍵字

液化澱粉芽孢桿菌 ； 果膠酶 ； 麥麩

並列摘要

Pectin-containing by products possess potential applications in many fields. In this research, different kinds of agricultural pectin-containing by products from banana (banana peel), rice (rice bran), orange (orange peel), coffee (spent coffee ground), as well as wheat (wheat bran) were used to provide both carbon and nitrogen (C/N) nutrients for the production of a pectinase via Bacillus amyloliquefaciens TKU050, a pectinolytic bacterium isolated from the soils of Tamsui. B. amyloliquefaciens TKU050 expressed the highest pectinase productivity (0.76 U/mL) on 0.5% wheat bran-containing medium at 37 ◦C for 4 days. After purification by ammonium sulfate precipitation, High Q column chromatography, and HPLC, a pectinase (TKU050) was purified from the culture supernatant fermented under optimized culture conditions. The molecular mass of TKU050 pectinase was estimated to be 58 kDa by SDS-PAGE. The recovery ratio and specific activity of TKU050 pectinase were 7.24% and 0.51 U/mg, respectively. The optimum temperature, optimum pH, thermal stability, and pH stability of TKU050 pectinase were 50◦C, pH6, <50◦C, and pH 6-9, respectively. TKU050 pectinase was inhibited Cu2+ and SDS. The reducing sugar obtained by hydrolyzing with TKU050 pectinase showed enhancing effect on the growth of lactobacillus acid bacteria.

並列關鍵字

Bacillus amyloliquefaciens ； pectinase ； wheat bran

參考文獻

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