枯草桿菌 TKU049 所生產果膠酶的純化與定性

Purification and characterization of a pectinase from Bacillus subtilis TKU049

謝孝婷(Hsiao-Ting Hsieh)

指導教授 ： 王三郎

共同指導教授 ： 阮文邦(Van-Bon Nguyen)

淡江大學/理學院/化學學系碩士班/碩士(2020年)

https://doi.org/10.6846/TKU.2020.00899

摘要

由於果膠酶的重要生物技術用途，在食品和飲料行業具有廣闊的工業前景。本研究使用的菌株Bacillus subtilis TKU049係從淡江大學土壤中篩選出來，探討香蕉皮、橙皮、柚子皮、麥麩，米糠以及咖啡渣等六種農業副產品作為B. subtilis TKU049生產果膠酶之碳和氮的之可行性。 結果顯示於含有1.5％果膠的培養基中培養三天，B. subtilis TKU049所產生果膠酶活性最高（1.19 U / mL）。 SDS-PAGE分析純化所得TKU049果膠酶之分子量為60.5 kDa。 TKU049果膠酶之最適反應溫度及酸鹼值分別為60°C和pH 9，酸鹼穩定性則是pH6至pH10。
此外，於無機鹽對TKU049果膠酶酵素活性影響方面，Mn2+、Mg2+、Ca2+以及Fe2+金屬離子具促進酵素活性之作用。 而界面活性劑EDTA、Triton X-100、Tween 40以及Tween 20則對酵素活性影響不大。

關鍵字

枯草桿菌 ； 果膠 ； 果膠酶

並列摘要

Pectinases have tremendous industrial prospects to be used in the fields of food and beverage. In this study, pectin and six types of agricultural by-products (banana peel, orange peel, pomelo peel, wheat bran, rice bran, and coffee ground) were used as the sole carbon and nitrogen sources for pectinase production by Bacillus subtilis TKU049 which was isolated from the soil of Tamkang University. B. subtilis TKU049 produced highest pectinase activity of 1.19 U/mL when cultured for three days on a medium containing 1.5% of pectin. The molecular mass of TKU049 pectinase was determined to be 60.5 kDa by SDS-PAGE. TKU049 pectinase displayed optimal activity at 60 °C and pH 9, with a pH range of stability from 6 to 10. The enzyme activity of TKU049 pectinase was stimulated with Mn2+, Mg2+, Ca2+, and Fe2+ metal ions. Additionally, the TKU049 pectinase was stable in the presence of EDTA, Triton X-100, Tween 40, and Tween 20.

並列關鍵字

Bacillus subtilis ； pectin ； pectinase

參考文獻

1. Alaña, A., Alkorta, I., Domínguez, J. B., Llama, M. J., Serra, J. L. (1990). Pectin lyase activity in a Penicillium italicum strain. Applied and Environmental Microbiology, 56, 3755-3759.

2. Alimardani-Theuil, P., Gainvors-Claisse, A., Duchiron, F. (2011). Yeasts: an attractive source of pectinases—from gene expression to potential applications: a review. Process Biochemistry, 46, 1525-1537.

3. Alkorta, I., Garbisu, C., Llama, M. J., Serra, J. L. (1998). Industrial applications of pectic enzymes: a review. Process Biochemistry, 33, 21-28.

4. Angayarkanni, J., Palaniswamy, M., Murugesan, S., Swaminathan, K. (2002). Improvement of tea leaves fermentation with Aspergillus spp. pectinase. Journal of Bioscience and Bioengineering, 94, 299-303.

5. Beulah, D., Sunitha, E., Srilakshmi, T. (2015). Production, purification and assay of pectinase enzyme from Aspergillus niger. Helix, 1, 673-645.