摘要
采用絮凝、超滤和冷冻干燥组合的方法制备重组枯草芽孢杆菌亮氨酸氨肽酶,以氨肽酶切除末端疏水性氨基酸为酶解脱苦效果的指标,考察了该氨肽酶与脯氨酸氨肽酶协同水解大米肽的脱苦效果。酶的制备工艺条件如下:加入0.15%±0.008%的絮凝剂,2.5%±0.006%的硅藻土,调节p H8.0进行过滤,采用30 k Da的PES卷式膜,超滤浓缩7倍,氨肽酶的总回收率为64.69%±1.29%;在优化的酶解条件下,大米肽的酶解液中的亮氨酸、精氨酸及游离的疏水性氨基酸总量分别是酶解前的3.39±0.10、16.48±0.49、4.39±0.13倍,酶解后分子量500~1000 Da的肽含量降低了30.63%±0.61%,经双酶协同水解后脯氨酸含量是单酶水解后的1.78±0.07倍。优化后工艺简便,且该酶在大米肽脱苦中有良好的应用。
The combination of fiocculation, uhrafihration, and freeze drying method to extract the recombinant bacillus subtilis leucine aminopeptidase,with aminopeptidase removal terminal hydrophobic amino acids for debittering effect index, debittering effect effects of leucine aminopeptidase and proline aminopeptidase hydrolysis of rice peptide was investigated.The preparation conditions of the enzyme were as follows：adding 0.15% ± 0.008% floeeulant,2.5% ± 0.006% diatomite, adjusting pH8.0 for filtration,using 30 kDa PES roll membrane, ultrafiltration concentration 7 times, under this condition ： the total recovery of aminopeptidase rate was 64.69% ± 1.29%. In the optimized enzymatic hydrolysis conditions, the total amount of leucine, arginine and free hydrophobic amino acids in the hydrolyzate were 3.39 ± 0.10,16.48 ± 0.49 and 4.39 ± 0.13 times higher than that of the hydrolyzate.After the enzymolysis, the molecular weight was 500-1000 Da was reduced by 30.63 %± 0.61%, and the content of proline was 1.78±0.07 times higher than that of single enzyme hydrolysis after double enzyme synergistic hydrolysis. The ontimized process is simple and the enzyme has Rood application in the peeling of rice peptides.
出处
《食品工业科技》
CAS
CSCD
北大核心
2017年第19期109-113,共5页
Science and Technology of Food Industry
基金
江苏省产学研联合创新资金(BY2014023-22)
关键词
亮氨酸氨肽酶
提取
回收率
协同水解
疏水性氨基酸
leucine aminopeptidase
extraction
recovery rate
synergistic hydrolysis
hydrophobic aminoacid
