摘要
利用溶胶-凝胶原理,以正硅酸乙酯(TEOS)为原料,3-缩水甘油醚氧丙基三甲氧基硅烷(KH-560)为偶联剂,在酸性条件下发生缩合反应制备新型基质材料,通过红外光谱(FT-IR),固体核磁共振波谱(CP/MAS13C NMR)和X-射线衍射(XRD)对基质载体进行结构表征。通过交联-包埋固定化法制备新型纤维素固载酶,分析研究了固载酶及游离酶酶解不同微观尺度纤维素基质的酶学性能。结果表明:固载酶进行酶促反应的最优p H 4.0,最优温度为60℃,表明固载酶的热稳定性优于游离酶;测得固载酶的米氏常数与游离酶无显著差异;固定化纤维素酶的重复使用性和储存稳定性较游离酶均有较大改善。有关研究可为制备性能优良的固载酶提供一定借鉴。
A novel carrier was synthesized by condensation reaction with an acid catalytic action using tetraethyl orthosilicate and KH-560 as the silica resource and coupling reagent, respectively. The composition and structure of the carrier were characterized by infrared absorption spectroscopy,13 C nuclear magnetic resonance, and X-ray diffractometry. The novel immobilized cellulase was prepared by the crosslinking-embedding immobilized method. The enzymatic characteristics of the immobilized cellulase were tested and compared with that of free cellulase. Experimental results showed that the immobilized cellulase maintained higher activity in the broader ranges of p H and temperature. The Michaelis constant(Km) of immobilized cellulase(1.58 g·L-1)is similar as the free cellulase(1.40 g·L-1). Thus, immobilized cellulase could have a stronger affinity,reusability, and storage stability; and could provide a reference for preparing immobilized enzymes to perform better.
出处
《浙江农林大学学报》
CAS
CSCD
北大核心
2016年第2期315-321,共7页
Journal of Zhejiang A&F University
基金
浙江省大学生科技创新活动计划暨新苗人才计划资助项目(2014R412026
201111030107)
关键词
纤维素酶
交联-包埋
固定化
催化性能
cellulase
crosslinking and embedding
immobilized
catalytic property
