从山榄科植物赛莨菪种子中纯化得到的一种 Bowman-Birk 型胰蛋白酶抑制剂的特性研究。
Characterization of a Bowman-Birk type trypsin inhibitor purified from seeds of Solanum surattense.
机构信息
Department of Biotechnology, Shivaji University, Kolhapur, MS, 416004, India.
Department of Biochemistry, School of Life Sciences, Kavayitri Bahinabai Chaudhari North Maharashtra University, Jalgaon, MS, 425001, India.
出版信息
Sci Rep. 2021 Apr 21;11(1):8648. doi: 10.1038/s41598-021-87980-8.
A Bowman-Birk type trypsin inhibitor protein (SSTI) from seeds of the medicinal plant Solanum surattense was isolated, purified and characterized. SSTI showed a single band on SDS-PAGE corresponding to 11.4 kDa molecular weight. It is a glycoprotein (2.8% glycosylation) that differentially interacted with trypsin and chymotrypsin in a concentration-dependent manner. Its peptide sequence is similar to other Bowman-Birk type protease inhibitors found in Glycine max and Phaseolus acutifolius. The inhibitory activity was stable over a wide range of pH (1-10) and temperatures (10-100° C). Far-UV Circular Dichroism (CD) studies showed that SSTI contains β sheets (~ 23%) and α helix (~ 6%) and demonstrated structural stability at wide pH and high temperature. The kinetic analysis revealed a noncompetitive (mixed) type nature of SSTI and low inhibitor constant (Ki) values (16.6 × 10 M) suggested strong inhibitory activity. Isothermal titration calorimetric analysis revealed its high affinity towards trypsin with dissociation constant (K) 2.28 µM.
从药用植物酸浆（Solanum surattense）的种子中分离、纯化并表征了一种 Bowman-Birk 型胰蛋白酶抑制剂蛋白（SSTI）。SSTI 在 SDS-PAGE 上显示出单一条带，对应于 11.4 kDa 的分子量。它是一种糖蛋白（2.8%的糖基化），以浓度依赖的方式与胰蛋白酶和糜蛋白酶不同相互作用。其肽序列与在 Glycine max 和 Phaseolus acutifolius 中发现的其他 Bowman-Birk 型蛋白酶抑制剂相似。抑制活性在广泛的 pH（1-10）和温度（10-100°C）范围内稳定。远紫外圆二色性（CD）研究表明，SSTI 含有 β 片层（23%）和 α 螺旋（6%），并在宽 pH 和高温下表现出结构稳定性。动力学分析表明 SSTI 具有非竞争性（混合）类型性质，抑制剂常数（Ki）值较低（16.6×10^-6 M），表明其具有很强的抑制活性。等温滴定量热法分析表明，它与胰蛋白酶具有高亲和力，解离常数（K）为 2.28 µM。
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