Preparation and characterization of a highly stable phenoxazinone synthase nanogel.
Preparation and characterization of a highly stable phenoxazinone synthase nanogel.
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DOI：
10.1186/s13065-016-0178-8
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发表时间：
2016
影响因子：
--
通讯作者：
Wei P
中科院分区：
文献类型：
--
作者：
Jia H;Gao Z;Ma Y;Zhong C;Wang C;Zhou H;Wei P
吩恶嗪酮合酶（PHS）是一种来源于链霉菌的漆酶样多铜氧化酶，具有很大的工业应用潜力。在本文中，我们制备的PHS纳米凝胶保留其初始活性的82%，通过水溶液原位聚合在pH 9.3。基于动态光散射（DLS）分析，PHS纳米凝胶的平均直径为50.8nm。荧光分析表明，修饰后的酶分子结构的令人印象深刻的保存。PHS纳米凝胶在pH 4.0-4.5和50 °C下表现出最大的活性，而天然PHS的相应值为pH 4.5和40 °C。发现PHS纳米凝胶的Km和Vmax分别为0.052 mM和0.018 mM/min，而天然PHS的Km和Vmax分别为0.077 mM和0.021 mM/min。此外，PHS纳米凝胶具有更高的热稳定性、储存稳定性和耐溶剂性。PHS纳米凝胶的半衰期为1.71小时，与天然纳米凝胶的0.19小时相比增加了约9倍。综上所述，PHS纳米凝胶是一种很有前途的生物催化剂。
Phenoxazinone synthase (PHS) is a laccase-like multicopper oxidase originating from Streptomyces with great industrial application potential. In this paper, we prepared the PHS nanogel retaining 82 % of its initial activity by aqueous in situ polymerization at pH 9.3. The average diameter of the PHS nanogel was 50.8 nm based on dynamic light scattering (DLS) analysis. Fluorescence analysis indicated the impressive preservation of the enzyme molecular structure upon modification. The PHS nanogel exhibited the most activity at pH 4.0–4.5 and 50 °C while the corresponding values were pH 4.5 and 40 °C for the native PHS. The Km and Vmax of the PHS nanogel were found to be 0.052 mM and 0.018 mM/min, whereas those of the native PHS were 0.077 mM and 0.021 mM/min, respectively. In addition, the PHS nanogel possessed higher thermal and storage stability and solvent tolerance compared with the native one. The half-life of the PHS nanogel was 1.71 h and multiplied around ninefold compared to 0.19 h for the native one. In summary, the PHS nanogel could be a promising biocatalyst in industry.