壳聚糖脱乙酰酶的表达和分子修饰。
Expression and Molecular Modification of Chitin Deacetylase from .
机构信息
Qingdao Key Laboratory of Food Biotechnology, College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China.
Key Laboratory of Biological Processing of Aquatic Products, China National Light Industry, Qingdao 266404, China.
出版信息
Molecules. 2022 Dec 23;28(1):113. doi: 10.3390/molecules28010113.
Chitin deacetylase can be used in the green and efficient preparation of chitosan from chitin. Herein, a novel chitin deacetylase CDA from was heterologously expressed and comprehensively characterized. DNA exhibits its highest deacetylation activity at 35 °C and pH 8.0. The enzyme activity is enhanced by Mn and prominently inhibited by Zn, SDS, and EDTA. CDA showed better deacetylation activity on colloidal chitin, (GlcNAc), and (GlcNAc) than other forms of the substrate. Molecular modification of CDA was conducted based on sequence alignment and homology modeling. A mutant CDA63G with higher activity and better temperature stability was obtained. The deacetylation activity of CDA63G was increased by 133% compared with the original enzyme, and the optimal reaction temperature increased from 35 to 40 °C. The half-life of CDA63G at 40 °C is 15 h, which was 5 h longer than that of the original enzyme. The improved characteristics of the chitin deacetylase CDA63G make it a potential candidate to industrially produce chitosan from chitin.
壳聚糖脱乙酰酶可用于从甲壳素绿色高效制备壳聚糖。本文从 中异源表达并全面表征了一种新型的壳聚糖脱乙酰酶 CDA。DNA 在 35°C 和 pH 8.0 时表现出最高的脱乙酰化活性。该酶的活性被 Mn 增强，被 Zn、SDS 和 EDTA 显著抑制。CDA 对胶体甲壳素、(GlcNAc) 和 (GlcNAc) 的脱乙酰化活性优于其他形式的底物。基于序列比对和同源建模对 CDA 进行了分子修饰。获得了活性更高、热稳定性更好的突变体 CDA63G。与原始酶相比，CDA63G 的脱乙酰化活性提高了 133%，最佳反应温度从 35°C 升高到 40°C。CDA63G 在 40°C 时的半衰期为 15 小时，比原始酶长 5 小时。壳聚糖脱乙酰酶 CDA63G 的这些改进特性使其成为从甲壳素工业生产壳聚糖的潜在候选酶。
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