蒸汽爆破预处理联合壳聚糖酶制备壳三糖  

Chitotriose Preparation by Steam Explosion Pretreatment and Chitosanase

作　　者：郭超然 赵红军 苏海鹏 黄海燕 孙建安[1] 毛相朝[1] GUO Chaoran;ZHAO Hongjun;SU Haipeng;HUANG Haiyan;SUN Jianan;MAO Xiangzhao(College of Food Science and Engineering,Ocean University of China,Qingdao 266003,Shandong,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《食品研究与开发》2023年第14期192-197,共6页Food Research and Development

基　　金：国家自然科学基金项目(U21A20271);山东省重点研发计划项目(2022TZXD001);国家虾蟹产业技术体系建设项目(CARS-48)。

摘　　要：为解除壳聚糖底物的不溶解性对壳聚糖酶的限制,并实现单一聚合度壳寡糖的绿色制备,该研究对壳聚糖进行蒸汽爆破预处理,随后使用在枯草芽孢杆菌WB800中高效表达的壳聚糖酶Csn-But进行酶解。扫描电镜结果显示,经过2.4 MPa蒸汽爆破处理的壳聚糖显示多刺的纤维状细枝排列,且表面被明显撕裂,分离和暴露的纤维中含有多个空腔。傅里叶变换衰减全反射红外光谱结果显示蒸汽爆破处理后壳聚糖的化学键未被破坏,氢键网络被破坏。使用壳聚糖酶Csn-But酶解在2.4 MPa蒸汽爆破压力下处理的壳聚糖48 h后,壳寡糖含量较未预处理对照组提高5.4倍,产物浓度达到2.0 mg/mL。并通过调控底物浓度,在作用6%浓度的底物时,实现了高纯度壳三糖的制备。To overcome the limitation of the insolubility of chitosan substrates to chitosanase and achieve the green preparation of chitooligo saccharides with a single degree of polymerization (DP),this study conducted steam explosion pretreatment on chitosan and performed enzymatic hydrolysis by using chitosanase Csn-But,which was efficiently expressed in Bacillus subtilis WB800.Scanning electron microscopy (SEM) results showed that the chitosan treated with the steam explosion at 2.4 MPa exhibited a multi-spiked fibrous arrangement and obvious tearing on the surface,with multiple cavities present in the separated and exposed fibers.The results of attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy showed that the chemical bonds of chitosan were not destroyed by steam explosion treatment,but the hydrogen bonding network was disrupted.After using chitosanase Csn-But to hydrolyze the chitosan treated with the steam explosion (2.4 MPa) for 48 h,the content of chitooligosaccharides was 5.4 times higher than that of the control group without pretreatment,with a product concentration of 2.0 mg/mL.Furthermore,by regulating the substrate concentration,high-purity chitotriose was obtained from the substrate with a concentration of 6%.

关 键 词：壳聚糖酶 枯草芽孢杆菌 粉状壳聚糖 蒸汽爆破 壳三糖 

分 类 号：TQ426.97[化学工程] O636.1[理学—高分子化学]