Key Laboratory of Molecular Biophysics, Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan, China.
Appl Biochem Biotechnol. 2012 Feb;166(4):925-32. doi: 10.1007/s12010-011-9480-z. Epub 2011 Dec 15.
Cross-linking of enzyme aggregates from recombinant Geotrichum sp. lipase based on polyethylenimine (PEI) was applied to hydrolyze fish oil for enrichment of polyunsaturated fatty acids successfully. Through acetone precipitation and cross-linking of physical aggregates using glutaraldehyde in the presence of PEI, firmly cross-linked enzyme aggregates (PEI-CLEAs) were prepared. They could maintain more than 65% of relative hydrolysis degree after incubation in the range of 50-55 °C for 4 h and maintain more than 85% of relative hydrolysis degree after being treated by acetone, tert-butyl alcohol and octane for 4 h. PEI-CLEAs increased hydrolysis degree to 42% from 12% by free lipase. After five batch reactions, PEI-CLEAs still maintained 72% of relative hydrolysis degree. Hydrolysis of fish oil by PEI-CLEAs produced glycerides containing concentrated EPA and DHA in good yield. PEI-CLEAs had advantages over general CLEAs and free lipase in initial reaction rate, hydrolysis degree, thermostability, organic solvent tolerance and reusability.
基于聚乙烯亚胺（PEI）交联重组根霉脂肪酶的酶聚集体，成功地用于水解鱼油以富集多不饱和脂肪酸。通过在存在 PEI 的情况下使用戊二醛沉淀和物理聚集体交联，制备了牢固交联的酶聚集体（PEI-CLEAs）。它们在 50-55°C 的范围内孵育 4 小时后，仍能保持超过 65%的相对水解度，并且在经过丙酮、叔丁醇和辛烷处理 4 小时后，仍能保持超过 85%的相对水解度。PEI-CLEAs 使游离脂肪酶的水解度从 12%提高到 42%。经过五次分批反应后，PEI-CLEAs 仍保持 72%的相对水解度。PEI-CLEAs 用于水解鱼油，可高产率地生成含有浓缩 EPA 和 DHA 的甘油酯。与普通 CLEAs 和游离脂肪酶相比，PEI-CLEAs 在初始反应速率、水解度、热稳定性、有机溶剂耐受性和可重复使用性方面具有优势。
