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Mining and Expression of a Metagenome-Derived Keratinase Responsible for Biosynthesis of Silver Nanoparticles
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2018-02-12 00:00:00 , DOI: 10.1021/acsbiomaterials.7b00687 Li-Yan Tao 1 , Jin-Song Gong 1 , Chang Su 1 , Min Jiang 1 , Heng Li 1 , Hui Li 1 , Zhen-Ming Lu 1, 2 , Zheng-Hong Xu 1, 2 , Jin-Song Shi 1
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2018-02-12 00:00:00 , DOI: 10.1021/acsbiomaterials.7b00687 Li-Yan Tao 1 , Jin-Song Gong 1 , Chang Su 1 , Min Jiang 1 , Heng Li 1 , Hui Li 1 , Zhen-Ming Lu 1, 2 , Zheng-Hong Xu 1, 2 , Jin-Song Shi 1
Affiliation
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A keratinase gene kerBv was mined from soil metagenomes. The open reading frame consisted of 1149 bp and potentially encoded a protein of 382 amino acid residues. It shared the same active site with several reported typical keratinases via analysis of the amino acid sequence. The keratinase was successfully expressed in B. subtilis WB600 with pMA5 expression vector. The maximum activity of 164.8 U/mL in the fermentation supernatant was observed after incubating for 30 h in Terrifc Broth (TB) medium. The keratinase exhibited outstanding resistance to metal ions and was surfactant-stable. Additionally, the enzyme displayed broad substrate specificity especially toward insoluble substrate feather meal because of its disulfide bond-reducing activity. Furthermore, the reducing power of the recombinant keratinase was investigated. It showed that the protein exhibited a relatively high reducing power, which was subsequently used in the biosynthesis of silver nanoparticles (AgNPs). The biosynthesized AgNPs were characterized by ultraviolet–visible (UV–vis) spectroscopy, dynamic light scattering (DLS), transmission electron microscope (TEM), as well as Fourier transform infrared spectroscopy (FTIR) and displayed obvious antibacterial activities toward Escherichia coli.
中文翻译：
银基因纳米粒子的生物合成负责的基因组基因角蛋白酶的提取和表达。
角蛋白酶基因kerBv是从土壤基因组中提取的。开放阅读框由1149 bp组成，并可能编码382个氨基酸残基的蛋白质。通过对氨基酸序列的分析，它与几种报道的典型角蛋白酶共享相同的活性位点。角蛋白酶在枯草芽孢杆菌中成功表达带有pMA5表达载体的WB600。在Terrifc Broth（TB）培养基中孵育30小时后，观察到发酵上清液的最大活性为164.8 U / mL。角蛋白酶显示出对金属离子的优异抗性并且是表面活性剂稳定的。另外，由于其二硫键还原活性，该酶表现出广泛的底物特异性，特别是对不溶性底物羽毛粉。此外，研究了重组角蛋白酶的还原能力。结果表明，该蛋白表现出较高的还原能力，随后被用于银纳米颗粒（AgNPs）的生物合成中。生物合成的AgNPs的特征在于紫外可见光谱（UV-vis），动态光散射（DLS），透射电子显微镜（TEM），大肠杆菌。
更新日期：2018-02-12
中文翻译：
银基因纳米粒子的生物合成负责的基因组基因角蛋白酶的提取和表达。
角蛋白酶基因kerBv是从土壤基因组中提取的。开放阅读框由1149 bp组成，并可能编码382个氨基酸残基的蛋白质。通过对氨基酸序列的分析，它与几种报道的典型角蛋白酶共享相同的活性位点。角蛋白酶在枯草芽孢杆菌中成功表达带有pMA5表达载体的WB600。在Terrifc Broth（TB）培养基中孵育30小时后，观察到发酵上清液的最大活性为164.8 U / mL。角蛋白酶显示出对金属离子的优异抗性并且是表面活性剂稳定的。另外，由于其二硫键还原活性，该酶表现出广泛的底物特异性，特别是对不溶性底物羽毛粉。此外，研究了重组角蛋白酶的还原能力。结果表明，该蛋白表现出较高的还原能力，随后被用于银纳米颗粒（AgNPs）的生物合成中。生物合成的AgNPs的特征在于紫外可见光谱（UV-vis），动态光散射（DLS），透射电子显微镜（TEM），大肠杆菌。
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