Key Laboratory of Ministry of Education Industrial Fermentation Microbiology, Tianjin Key Laboratory of Industrial Microbiology, Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control, College of Biotechnology, Tianjin University of Science and Technology , Tianjin, P. R. China.
Bioengineered. 2021 Dec;12(1):266-277. doi: 10.1080/21655979.2020.1869438.
Chitosan hydrolysis by chitosanase is one of the most effective methods to produce chitosan oligosaccharides. One of the prerequisites of enzyme fermentation production is to select and breed enzyme-producing cells with good performance. So in the process of fermentation production, the low yield of chitosanase cannot meet the current requirement. In this paper, a strain producing chitosanase was screened and identified, and a novel mutagenesis system (Atmospheric and Room Temperature Plasma (ARTP)) was selected to increase the yield of chitosanase. Then, the fermentation medium was optimized to further improve the enzyme activity of the strain. A strain of capable of producing chitosanase was screened and identified from soil samples. A mutant strain of was obtained by Atmospheric and Room Temperature Plasma mutagenesis and bioscreening method, and chitosanase activity was 2.49 folds that of the original bacterium. After an optimized fermentation medium, the enzyme activity of the mutant strain was 1.47 folds that of the original bacterium. Combined with all the above optimization experiments, the enzyme activity of mutant strain increased by 3.66 times. The results showed that the Atmospheric and Room Temperature Plasma mutagenesis and bioscreening method could significantly increase the yield of chitosanase in , and had little effect on the properties of the enzyme. These findings have potential applications in the mutagenesis of other enzyme-producing microorganisms.
壳聚糖酶水解壳聚糖是生产壳寡糖最有效的方法之一。酶发酵生产的前提之一是筛选和培育具有良好性能的产酶细胞。因此,在发酵生产过程中,壳聚糖酶的低产量不能满足当前的要求。本文筛选鉴定了一株产壳聚糖酶的菌株,并选择了一种新型的诱变系统(大气压室温等离子体(ARTP))来提高壳聚糖酶的产量。然后,对发酵培养基进行了优化,以进一步提高菌株的酶活性。从土壤样品中筛选鉴定出一株产壳聚糖酶的菌株。通过大气压室温等离子体诱变和生物筛选法获得了一株壳聚糖酶活性提高了 2.49 倍的突变株。经过优化发酵培养基,突变株的酶活是原始菌株的 1.47 倍。结合以上所有优化实验,突变株的酶活提高了 3.66 倍。结果表明,大气压室温等离子体诱变和生物筛选法可显著提高 产壳聚糖酶的产量,而对酶的性质影响较小。这些发现可能在其他产酶微生物的诱变中有应用前景。