壳聚糖固定β-淀粉酶-海藻糖合酶双功能融合酶产海藻糖的研究  

Immobilization of β-amylase-trehalose synthase bifunctional fusion enzyme by chitosan

作　　者：郑海杰[1] 徐波[1] 苏静[1] 王瑞明[1] 马春玲[1] 

机构地区：[1]齐鲁工业大学生物工程学院山东省微生物重点实验室,济南250353

出　　处：《食品科技》2017年第7期244-248,共5页Food Science and Technology

基　　金：国家自然科学基金项目(31401626)

摘　　要：β-淀粉酶-海藻糖合酶双功能融合酶以戊二醛为交联剂,壳聚糖为载体,采用交联-吸附法固定。通过单因素试验和正交设计方案得到最优条件:壳聚糖浓度为2.5 g/L,戊二醛浓度3 g/L,交联时间80 min,融合酶浓度为8 mg/m L,固定时间为12 h,酶活力回收率为71%。酶学特性:最适pH向碱性偏移0.5,最适温度不变,具有良好的储存稳定性和操作稳定性,其酸碱稳定性和热稳定性均优于游离态。4℃储存31 d后酶活力损失33%,循环使用6次酶活力降低27%,通过固定化海藻糖转化效率与游离态和单酶混合体系相比分别低29%、6%。β-amylase-trehalose synthase fusion enzyme was immobilized by cross-linking and adsorption method with glutaraldehyde as crosslinking agent and chitosan as carrier. The optimum conditions were obtained by single factor test and orthogonal design： the concentration of chitosan was 2.5 g/L, the concentration of glutaraldehyde was 3 g/L, the crosslinking time was 80 min, the concentration of fusion enzyme was 8 mg/mL, the immobilization was 12 h, the recovery rate of fusion enzyme activity was 71%. Enzymatic differences in characteristics： optimum pH to alkaline offset 0.5, optimum temperature constant, had good storage stability and operational stability, while its pH stability and thermal stability were better than the free state. The enzyme activity was reduced by 33% and 27%, after storage 31 d at 4 ℃ and 6 cycles, respectively. The immobilized trehalose conversion efficiency was 29% and 6% lower than that of the free and single enzyme mixtures, respectively.

关 键 词：β-淀粉酶-海藻糖合酶双功能融合酶 壳聚糖 戊二醛 固定化酶 

分 类 号：TS201.25[轻工技术与工程—食品科学]