来自黄色粘球菌的一种新型酸性脂氧合酶的表达、纯化及特性分析
Expression, purification, and characterization of a novel acidic Lipoxygenase from Myxococcus xanthus.
作者信息
Qian Hui, Xia Bingjie, He Yujun, Lu Zhaoxin, Bie Xiaomei, Zhao Haizhen, Zhang Chong, Lu Fengxia
机构信息
College of Food Science and Technology, Nanjing Agricultural University, Weigang, Nanjing 210095, PR China.
College of Food Science and Technology, Nanjing Agricultural University, Weigang, Nanjing 210095, PR China.
出版信息
Protein Expr Purif. 2017 Oct;138:13-17. doi: 10.1016/j.pep.2017.05.006. Epub 2017 May 25.
The gene encoding a novel acidic lipoxygenase from Myxococcus xanthus DK1622 (accession: WP_011551853.1) was cloned into vector pET-28a and expressed in Escherichia coli BL21(DE3). The recombinant enzyme (rMxLOX), with a molecular weight of approximately 80 kDa, was purified to homogeneity using one-step nickel-affinity chromatography and showed an activity of 5.6 × 10 U/mg. The optimum pH and temperature for rMxLOX activity were found to be 3.0 and 30 °C, respectively. Purified rMxLOX exhibited activity towards linoleic acid and arachidonic acid as substrates, with linoleic acid being the better substrate (K and k values of 0.048 mM and 13.3/s, respectively). The synthetic dye aniline blue was decolorized 69.7 ± 3.5%, following incubation with rMxLOX for 35 min. These results reveal the potential for the use of rMxLOX in the pulp, textile, and wastewater treatment industries.
编码来自黄色黏球菌DK1622的一种新型酸性脂氧合酶的基因（登录号：WP_011551853.1）被克隆到载体pET-28a中，并在大肠杆菌BL21(DE3)中表达。重组酶（rMxLOX）的分子量约为80 kDa，通过一步镍亲和层析纯化至同质，其活性为5.6×10 U/mg。发现rMxLOX活性的最适pH和温度分别为3.0和30℃。纯化的rMxLOX以亚油酸和花生四烯酸为底物表现出活性，其中亚油酸是更好的底物（K和k值分别为0.048 mM和13.3/s）。与rMxLOX孵育35分钟后，合成染料苯胺蓝的脱色率为69.7±3.5%。这些结果揭示了rMxLOX在制浆、纺织和废水处理行业应用的潜力。
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