Coimmobilization of beta-Agarase and alpha-Neoagarobiose Hydrolase for Enhancing the Production of 3,6-Anhydro-L-galactose
Coimmobilization of beta-Agarase and alpha-Neoagarobiose Hydrolase for Enhancing the Production of 3,6-Anhydro-L-galactose
复制标题
共固定 β-琼脂酶和 α-新琼脂二糖水解酶以提高 3,6-脱水-L-半乳糖的产量
DOI：
10.1021/acs.jafc.8b01974
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发表时间：
2018
影响因子：
6.1
通讯作者：
Mao Xiangzhao
中科院分区：
文献类型：
--
作者：
Wang Qidong;Sun Jianan;Liu Zhen;Huang Wencan;Xue Changhu;Mao Xiangzhao
在此，我们报道了一种简单有效的方法，通过具有共固定化 β-琼脂酶 AgWH50B 和 α-新琼脂二糖水解酶 K134D 的多酶系统一步生产 3,6-脱水-L-半乳糖 (l-AHG) 和琼脂三糖 (AO3)。 K134D 通过 AgaWH117 诱变获得，当通过共价键固定在功能化磁性纳米颗粒上时，显示出更高的热稳定性。通过傅里叶变换红外光谱（FTIR）对所得多酶生物催化剂进行了表征。与游离琼脂酶相比，共固定化琼脂酶表现出相对较高的琼脂糖至l-AHG的转化效率。共固定化琼脂酶得到的l-AHG产率为40.6%，比游离琼脂酶提高了6.5%。八个循环后，多酶生物催化剂仍保留初始活性的46.4%。据我们所知，这是第一份同时固定两种不同琼脂酶的报告。这些结果证明了通过共价键在磁性纳米粒子上制造新的多酶系统以产生1-AHG的新方法的可行性。
Here we report a simple and efficient method to produce 3,6-anhydro-l-galactose (l-AHG) and agarotriose (AO3) in one step by a multienzyme system with the coimmobilized β-agarase AgWH50B and α-neoagarobiose hydrolase K134D. K134D was obtained by AgaWH117 mutagenesis and showed improved thermal stability when immobilized via covalent bonds on functionalized magnetic nanoparticles. The obtained multienzyme biocatalyst was characterized by Fourier transform infrared spectroscopy (FTIR). Compared with free agarases, the coimmobilized agarases exhibited a relatively higher agarose-to-l-AHG conversion efficiency. The yield ofl-AHG obtained with the coimmobilized agarases was 40.6%, which was 6.5% higher than that obtained with free agarases. After eight cycles, the multienzyme biocatalyst still preserved 46.4% of the initial activity. To the best of our knowledge, this is the first report where two different agarases were coimmobilized. These results demonstrated the feasibility of the new method to fabricate a new multienzyme system onto magnetic nanoparticles via covalent bonds to producel-AHG.