通过重组黑曲霉HEWL WT-13-16提高鸡蛋清溶菌酶（HEWL）产量的生物加工策略

Bioprocessing strategies for improving hen egg-white lysozyme (HEWL) production by recombinant Aspergillus niger HEWL WT-13-16.

作者信息

Gyamerah M, Merichetti G, Adedayo O, Scharer J M, Moo-Young M

机构信息

Department of Chemical Engineering, University of Waterloo, Ontario, N2L 3G1, Canada.

出版信息

Appl Microbiol Biotechnol. 2002 Dec;60(4):403-7. doi: 10.1007/s00253-002-1122-5. Epub 2002 Nov 5.

DOI:10.1007/s00253-002-1122-5

PMID:12466879

Abstract

Hen egg-white lysozyme (HEWL) production by recombinant Aspergillus niger HEWL WT-13-16 from a cDNA under the control of the A. niger glucoamylase promoter was used as a model system. The fungal mycelium was either immobilized on porous Celite 560 micro-carrier or grown in suspension as pelleted and dispersed forms. The objective was to reduce the protease activity that adversely affects the expressed HEWL. Free suspension culture at uncontrolled pH served as the benchmark. The control of pH during growth at pH 4.0 gave rise to a greater than five-fold reduction of protease activity in suspension culture. An additional 38.5% decrease in protease activity was achieved in mycelial-pellet cultures in comparison to a 40.9% decrease in protease activity obtained with Celite 560 beads in an airlift vessel at controlled pH. The specific HEWL yields were 5.8, 5.0 and 4.1 mg/g dry wt. for the free suspension, mycelial-pellet, and Celite-560-immobilized cultures, respectively.

摘要

以黑曲霉重组体HEWL WT-13-16在黑曲霉糖化酶启动子控制下由cDNA生产鸡蛋清溶菌酶（HEWL）作为模型系统。真菌菌丝体要么固定在多孔硅藻土560微载体上，要么以颗粒状和分散形式在悬浮液中生长。目的是降低对表达的HEWL产生不利影响的蛋白酶活性。在不受控制的pH值下进行的游离悬浮培养作为基准。在pH 4.0下生长期间控制pH值，使悬浮培养中的蛋白酶活性降低了五倍以上。与在控制pH值的气升式容器中使用硅藻土560珠获得的蛋白酶活性降低40.9%相比，菌丝体颗粒培养中的蛋白酶活性额外降低了38.5%。游离悬浮培养、菌丝体颗粒培养和硅藻土560固定化培养的HEWL比产率分别为5.8、5.0和4.1 mg/g干重。