嗜热脂肪酶-无机杂化纳米花的制备及其性能研究  

Research about the preparation and characterisation of inorganic thermophilic lipase hybrid nanoflowers

作　　者：刘振山 时祎 陈剑雄 辛瑜 顾正华[1,2] 张梁 LIU Zhenshan;SHI Yi;CHEN Jianxiong;XIN Yu;GU Zhenghua;ZHANG Liang(National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]粮食发酵工艺与技术国家工程实验室(江南大学),江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与发酵工业》2022年第15期1-8,共8页Food and Fermentation Industries

基　　金：国家重点研发计划绿色生物制造专项(2021YF2100300)。

摘　　要：为满足工业生物催化需求,寻求一种简单、快捷、有效的脂肪酶固定化方法来扩大其实际应用范围。采用声化学方法,在10 min内将脂肪酶固定于由磷酸铜沉淀所构成的无机杂化纳米花中,并进一步通过扫描电子显微镜(scanning electron microscope,SEM)、X射线衍射(X-ray diffraction,XRD)和傅里叶红外光谱(Fourier transform infrared spectroscopy,FT-IR)等技术对制备的脂肪酶杂化纳米花进行表征。随后,研究了不同金属离子、金属盐离子类型、合成条件对脂肪酶杂化纳米花活性的影响。结果表明,Cu^(2+)可实现最佳固定化效果,且不同含铜无机盐类型对所形成的杂化纳米花活性存在影响。其中,使用CuSO_(4)合成纳米花可实现100%的包封率,而CuCl_(2)可使最终合成的纳米花相对酶活力达到145.7%。酶动力学研究结果表明,CuSO_(4)和CuCl_(2)纳米花的催化效率(k_(cat)/K_(m))分别是游离酶的106%和134%。此外,该固定化方法可有效提高固定化酶的储藏稳定性,在室温下储藏30 d其酶活力仍能保持在80%以上。在重复使用性研究中发现,固定化酶在连续使用5次后仍具有40%以上的催化活性。该研究结果有助于提高脂肪酶在工业生产中的适用性,并为其在生物柴油合成、洗涤剂及高温生产环境等多方面领域的应用奠定基础。Performed by enzyme,biocatalysis is considering as a green and sustainable technology that could eliminate the production of toxic and harmful by-products during the industrial manufacturing process.Lipase,which is believed as one of the most promising industrial enzymes that could catalysis the synthesis of lipid,interesterification,alcoholysis as well as acidolysis,has been characterized with advantages in catalytic activity,variety of substrates,regioselectivity and corresponding selectivity.However,limited by the inherent property of biomacromolecule,requirements including the high enzymatic activity,repeatability,storage stability and the feasibility of separation might not be meet easily in the industrial biocatalysis process when applying free enzyme without optimization.To fulfill the need in industrial application of bio-catalysis,it is desired to develop valid,simple as well as effective enzymatic immobilization method for improving the practical application potential of lipase.Recently,self-assembly organic-inorganic hybrid nanoflowers has been developed and applied as a novel enzymatic immobilization method.Via this technology,the limitations for the application of free enzymes could be overcome as the stability,repeatability and storage stability of the enzyme would be highly improved after immobilization.Nevertheless,it should be noted that such immobilizing process would normally involve an incubation period of three days at the temperature of 4℃.This relative slow preparation duration will bring considerable limitations for its further scale-up application,though both the enzymatic activity and stability could be significantly enhanced by immobilizing lipase in the form of nanoflowers.In order to break through the above restriction,sonochemistry was introduced in the current study to have the free lipase hybridized with metal phosphate precipitation within 10 min.In consideration of the potential effect of reaction conditions on the activity of the hybrid nanoflowers,parameters including the dif

关 键 词：脂肪酶 固定化 有机-无机杂化纳米花 催化活性 稳定性 

分 类 号：TQ426.97[化学工程] Q814.2[生物学—生物工程]