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首页> 外文会议>Proceedings of the 18th congress of the International Society for Mushroom Science >Purification and Characterization a Novel Alkaline Serine Protease from the Medicinal Mushroom Cordyceps sobolifera
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Purification and Characterization a Novel Alkaline Serine Protease from the Medicinal Mushroom Cordyceps sobolifera

机译：药用蘑菇冬虫夏草的新型碱性丝氨酸蛋白酶的纯化与鉴定

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摘要

A novel protease was purified from dried fruiting bodies of the mushroom Cordyceps sobolifera. The isolation procedure utilized ion exchange chromatography on DEAE-cellulose and SP-sepharose followed by gel filtration on Superdex 75. The protease did not adsorb on DEAE-cellulose but bound to SP-Sepharose. In sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE), the protease resolved as a single band with an apparent molecular mass of 31 kDa. It exhibited optimal activity at a temperature of 60 ℃, a pH of 12.4 with a Km of 1. 98 mg/mL and a Vmax of 23. 56 μg/mL/min against its substrate casein. Protease activity was enhanced by the Fe2+ion at low concentrations in 1. 25-10 mM and was strongly inhibited by Hg2+up to 1. 25 mM. The protease was strongly inhibited by phenylmethylsulfonyl fluoride (PMSF), suggesting that it was a serine protease. It manifested significant inhibitory activity toward HIV-1 reverse transcriptase (RT) with an IC50 value of 8. 2×10-3 μM, which was the highest anti-HIV-1 RT activity of reported mushroom proteins.
机译：从蘑菇冬虫夏草的干燥子实体中纯化了一种新的蛋白酶。分离方法是在DEAE-纤维素和SP-琼脂糖上进行离子交换层析，然后在Superdex 75上进行凝胶过滤。蛋白酶不吸附在DEAE-纤维素上，而是与SP-琼脂糖结合。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）中，蛋白酶分解为一条表带，表观分子量为31 kDa。它在60℃的温度，12.4的pH，1。98 mg / mL的Km和23. 56μg/ mL / min的Vmax方面表现出最佳活性。较低浓度下在1. 25-10 mM中的Fe2 +离子增强了蛋白酶的活性，而当Hg2 +浓度高达1. 25 mM时，其蛋白酶的活性受到强烈抑制。该蛋白酶被苯甲基磺酰氟（PMSF）强烈抑制，表明它是一种丝氨酸蛋白酶。它表现出对HIV-1逆转录酶（RT）的显着抑制活性，IC50值为8。2×10-3μM，是报道的蘑菇蛋白中最高的抗HIV-1 RT活性。

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  • 来源
  • 会议地点 Beijing(CN)
  • 作者

    Shou-Xian Wang; Guo-Qing Zhang; Shuang Zhao; Feng Xu; Lan-Qing Wang; Xiao-Li Geng; Li-Li Meng; Yu Liu; He-Xiang Wang;

  • 作者单位

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China;

    Key Laboratory of Urban Agriculture (North) of Ministry of Agriculture, Beijing University of Agriculture, Beijing 102206, China;

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;

    Institute of Plant Protection and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China;

    State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100193, China;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 菌类（食用菌）;
  • 关键词

    Alkaline Serine Protease; Fruiting Bodies; Mushroom; Cordyceps sobolifera;

    机译：碱性丝氨酸蛋白酶;果体;蘑菇;冬虫夏草;

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