Production of recombinant bovine lactoferrin N-lobe in insect cells and its antimicrobial activity

被引:27
作者
Nakamura, I
Watanabe, A
Tsunemitsu, H
Lee, NY
Kumura, H
Shimazaki, K
Yagi, Y
机构
[1] Hokkaido Univ, Fac Agr, Dairy Sci Lab, Kita Ku, Sapporo, Hokkaido 0608589, Japan
[2] Natl Inst Anim Hlth, Hokkaido res Stn, Lab Clin Biochem, Toyohira Ku, Sapporo, Hokkaido 0620045, Japan
[3] Natl Inst Anim Hlth, Shichinohe Res Unit, Shichinohe, Aomori 0392586, Japan
关键词
bovine lactoferrin; antimicrobial activity; baculovirus; insect cells; milk protein;
D O I
10.1006/prep.2001.1396
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Lactoferrin is a multifunctional, iron-binding glycoprotein found in physiological fluids of mammals. In the present study, a gene encoding the N-terminal half (N-lobe) of bovine lactoferrin was cloned and expressed in cultured insect cells using a baculovirus expression system. One mutation was found in the lactoferrin N-lobe gene, but it resulted in no amino acid substitution. The recombinant lactoferrin N-lobe was secreted into the culture medium and partially purified by means of an immobilized heparin column. The recombinant lactoferrin N-lobe secreted was not glycosylated, but it possessed antimicrobial activity toward Escherichia coli O111. The recombinant product synthesized and accumulated in the host cells exhibited greater electrophoretic mobility on SDS-PAGE than the secreted product and showed no potency to inhibit the growth of bacteria. It is thought that the product accumulated intracellularly lacks antimicrobial ability due to its degradation in the host cells or due to disruption of the active conformation. (C) 2001 Academic Press.
引用
收藏
页码:424 / 431
页数:8
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