Parasitology Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, India.
Int J Biol Macromol. 2018 Feb;107(Pt B):2375-2384. doi: 10.1016/j.ijbiomac.2017.10.114. Epub 2017 Oct 18.
Exploration of novel drug targets has been the major thrust area in filarial research. In this regard, identification and characterization of oxidative enzymes that play pivotal role in the survival of filarial parasite inside host are of immense importance. In this study, we are reporting the presence of an important redox regulatory enzyme, thioredoxin reductase (TrxR) in the bovine filarial parasite Setaria cervi. TrxR was found to be exists throughout the developmental stages viz. oocyte, microfilaria and adult of the parasite. Since further studies on this enzyme require adequate quantity, influential extraction parameters were optimized statistically using response surface methodology (RSM) employing a seven factors based Box-Behnken design matrix. ANOVA analysis revealed the relative importance of each parameter and a regression equation was eventually developed that could predict the specific activity (SA) of TrxR. Finally the optimized extraction conditions predicted by RSM was 6.1ml of 61.86mM buffer, pH 6.0, with extraction temperature 39.96°C for 180min in addition to 450rpm agitation and 20μl/ml of protease inhibitor. Therefore this study is going to be the maiden report depicting the identity of TrxR in filarial parasite and the optimized extraction conditions for its isolation with better kinetic efficiency.
新型药物靶点的探索一直是丝虫病研究的主要重点。在这方面，鉴定和表征在寄生虫宿主内生存中起关键作用的氧化酶至关重要。在这项研究中，我们报告了牛丝虫寄生虫 Setaria cervi 中存在一种重要的氧化还原调节酶，即硫氧还蛋白还原酶 (TrxR)。TrxR 存在于寄生虫的各个发育阶段，即卵母细胞、微丝蚴和成虫。由于对该酶的进一步研究需要足够的数量，因此使用基于七因素的 Box-Behnken 设计矩阵的响应面法 (RSM) 对有影响力的提取参数进行了统计学优化。方差分析揭示了每个参数的相对重要性，并最终开发出一个回归方程，可以预测 TrxR 的比活度 (SA)。最后，RSM 预测的优化提取条件为 6.1ml 61.86mM 缓冲液，pH 值为 6.0，提取温度为 39.96°C，持续 180min，同时搅拌速度为 450rpm，蛋白酶抑制剂添加量为 20μl/ml。因此，这项研究将首次报道 TrxR 在丝虫寄生虫中的存在，并优化其分离的提取条件，以提高动力学效率。
