| 粗毛栓菌木聚糖酶的纯化及性质 |
| Purification and characterization of xylanases from white-rot fungus Trametes gallica |
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| 中文关键词:白腐菌，同工酶，分离纯化，酶学性质 |
| 英文关键词:white-rot fungus, isoenzyme, xylanases, separation and purification, enzymatic properties |
| 基金项目:中国极地科学战略研究基金（No. 20080205）；国家自然科学基金（No. 30700107，No. 30500067） |
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| 中文摘要: |
| 以麦草粉为基质培养粗毛栓菌Trametes gallica，浸提固态培养物得浸提液，后经超滤浓缩、硫酸铵盐析、Phenyl-Sepharose CL-4B疏水层析、DEAE Sepharose fast flow阴离子交换层析和Sephadex G-150分子筛层析等分离与纯化步骤，获得部分纯化的木聚糖酶，其回收率和纯化倍数分别为1.45%和15.6。进一步经活性-PAGE回收，获得三种SDS-PAGE电泳纯级的木聚糖酶同工酶组分：XⅠ、XⅡ和XⅢ（按等电点从大到小排列）。三种组分分子量均约为19.0kDa；等电点分别为：5.6、4.7和4.0；含糖量分别为：0.25%、0.63%和3.4%；XⅠ既能降解木聚糖，又能降解纤维素；XⅡ的最适作用pH值为5.0，最适作用温度45℃；Mg2+、Fe2+对XⅡ有激活作用；Mn2+和Co2+有抑制作用；测得XⅡ的Km值为0.75mg/mL，Vmax 为5,000mmoL/min?mg。 |
| 英文摘要: |
| The white-rot fungus Trametes gallica was cultivated on wheat straw powder. Following protein precipitation with (NH4)2SO4, the fractions showing xylanase activity were separated and purified by chromatographic analyses with Phenyl-Sepharose CL-4B, DEAE Sepharose FF and Sephadex G-150. The recovery rate and purification fold reached up to 1.45% and 15.6-fold, respectively. The enzymes were further recovered by native-PAGE and three isoenzyme xylanases (XⅠ, XⅡ and XⅢ) of SDS-PAGE purity were finally obtained. The three components had similar molecular weights (approximately 19.0kDa), but exhibited different isolectric points (5.6 for XⅠ, 4.7 for XⅡ and 4.0 for XⅢ) and carbohydrate content (0.25% for XⅠ, 0.63% for XⅡ and 3.4% for XⅢ). XⅠ can degrade both xylan and cellulose, while XⅡ and XⅢ can only degrade xylan. The optimal temperature and pH for XⅡ were 45℃ and 5.0, respectively. Its activity was increased by Mg2+ and Fe2+, while inhibited by Mn2+ and Co2+. Its Km and Vmax were 0.75mg/mL and 5,000mmoL/min?mg, respectively. |
| 范美霞,付灿,孙迅,陶宗娅. 粗毛栓菌木聚糖酶的纯化及性质[J]. 菌物学报, 2010, 29(1): 83-90 |
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