磁性磷脂酶D交联酶聚集体的制备及其酶学性质  

Preparation and enzymatic properties of magnetic cross-linked phospholipase D aggregates

作　　者：石婷婷 吴蓉 杨猛 苏二正[1,2] SHI Tingting;WU Rong;YANG Meng;SU Erzheng(Co-innovation Center for the Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,Jiangsu,China;College of Light Industry and Food Engineering,Nanjing Forestry University,Nanjing 210037,Jiangsu,China)

机构地区：[1]南京林业大学,南方现代林业协同创新中心,江苏南京210037 [2]南京林业大学轻工与食品学院,江苏南京210037

出　　处：《精细化工》2023年第11期2463-2471,共9页Fine Chemicals

基　　金：江苏省第五期“333工程”培养资金资助项目(BRA2017458)。

摘　　要：以磁性Fe3O4为载体,采用吸附-聚集-交联的方法固定来源于Streptomyces chromofuscus的磷脂酶D(scPLD),制备了磁性磷脂酶D交联酶聚集体(MCLEA)。该方法制备的MCLEA酶活回收率可达72.89%,酶活为(437.00±6.60)U/g。与游离酶相比,MCLEA在不同温度和pH下的稳定性都得到了一定程度的提升,但由于与底物的亲和力降低,需要更高浓度的磷脂酰胆碱(PC)和Ca^(2+)作为反应底物与MCLEA结合。乙醇、四氢呋喃、叔丁醇、乙酸乙酯、乙醚和甲苯对MCLEA的酶促反应具有促进作用,MCLEA在进行连续13次酶促反应后,酶活仍可保留在76%以上,其半衰期为331.67min,该固定化方案可以提高酶的稳定性,从而提高其在催化反应过程中的重复利用性。Magnetic cross-linked phospholipase D from Streptomyces chromofuscus(scPLD)aggregates(MCLEA)were prepared by adsorption-aggregation-crosslinking with magnetic Fe3O4 as the carrier.The enzyme activity recovery of MCLEA was 72.89%,and the enzyme activity was(437.00±6.60)U/g.Compared to free scPLD,the MCLEA showed better tolerance at different temperatures and pH levels.However,due to the reduced affinity of MCLEA with substrate,higher concentrations of phosphatidylcholine(PC)and Ca^(2+)as reaction substrates were required for binding with MCLEA.Furthermore,organic solvents such as ethanol,tetrahydrofuran,tert-butanol,ethyl acetate,ether,and toluene could improve the enzymatic reaction of MCLEA.After 13 cycles of continuous enzymatic reaction,the activity of MCLEA remained above 76%,with a half-life of 331.67 min.This immobilization scheme could enhance the stability of the enzyme and thus improve its reusability in the catalytic reaction process.

关 键 词：磷脂酶D 吸附法 固定化 磁性磷脂酶D交联酶聚集体 性能 生物工程 

分 类 号：Q814.2[生物学—生物工程]