摘要
目的:研究重组内切几丁质酶的分离纯化及其催化壳聚糖制备壳寡糖的反应条件优化。方法:重组菌发酵液经PEG20000浓缩和DEAE-Sepharose FF离子交换层析后,测定总蛋白含量和内切几丁质酶活力。用纯化的内切几丁质酶催化壳聚糖制备壳寡糖,探讨其最佳的反应条件。结果:纯化的内切几丁质酶比活力41.34U/mg,纯化倍数2.49,酶总回收率89.63%。内切几丁质酶催化壳聚糖降解制备壳寡糖的最优化反应条件是:壳聚糖质量分数4%,p H7.0,温度30℃,时间12 h。结论:本研究结果为内切几丁质酶和壳寡糖的产业化应用奠定了良好基础。
Objective： To study the isolation and purification of the recombinant endochitinase and the optimization of the chito-oligosaccharide preparation by chitosan degradation using endochitinase. Methods： Variations in the total protein content and the endochitinase activity were determined after the recombinant fermentation broth was concentrated with PEG20000, and was chromatographed with DEAE-Sepharose FF ion-exchange column. The optimal reaction conditions of the chito- oligosaccharide preparation by chitosan degradation using endochitinase were explored. Results： The specific activity, the purification-fold and the recovery rate of the endochitinase obtained were 41.34 U/mg, 2.49 and 89.63%,respectively. The optimal reaction conditions of the chito-oligosaccharide preparation by chitosan degradation using endochitinase were as follows： chitosan concentration of 4%, p H 7.0, temperature 30 ℃ and time 12 h. Conclusion： The result lays a good foundation for the industrial application of endochitinase and chito-oligosaccharide.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2015年第2期99-103,共5页
Journal of Chinese Institute Of Food Science and Technology
基金
浙江省自然科学基金项目(M303087)
浙江省科学技术厅重点项目(2006C23073)
杭州市科技局项目(20061133B26)资助
关键词
内切几丁质酶
纯化
壳聚糖
反应条件优化
endochitinase
purification
chitosan
reaction condition optimization
