通过破坏wprA基因增强枯草芽孢杆菌中葡萄球菌激酶的分泌及细胞外稳定性。
Enhancement of secretion and extracellular stability of staphylokinase in Bacillus subtilis by wprA gene disruption.
作者信息
Lee S J, Kim D M, Bae K H, Byun S M, Chung J H
机构信息
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Republic of Korea.
出版信息
Appl Environ Microbiol. 2000 Feb;66(2):476-80. doi: 10.1128/AEM.66.2.476-480.2000.
Abstract
Staphylokinase (SAK), a polypeptide secreted by Staphylococcus aureus, is a plasminogen activator with a therapeutic potential in thrombosis diseases. A Bacillus subtilis strain which is multiply deficient in exoproteases was transformed by an expression plasmid carrying a promoter and a signal sequence of subtilisin fused in frame with the sak open reading frame. However, the amount of SAK secretion was marginal (45 mg/liter). In contrast, disruption of the wprA gene, which encodes a subtilisin-type protease, strongly promoted the production of SAK in the stationary phase (181 mg/liter). In addition, the extracellular stability of mature SAK was dramatically enhanced. These data indicate a significant role of the wprA gene product in degrading foreign proteins, both during secretion and in the extracellular milieu.
摘要
葡萄球菌激酶（SAK）是由金黄色葡萄球菌分泌的一种多肽，是一种纤溶酶原激活剂，在血栓性疾病中具有治疗潜力。用携带启动子和枯草杆菌蛋白酶信号序列且与sak开放阅读框读框融合的表达质粒转化一种外蛋白酶多重缺陷的枯草芽孢杆菌菌株。然而，SAK的分泌量很少（45毫克/升）。相比之下，编码枯草杆菌蛋白酶型蛋白酶的wprA基因的破坏在稳定期强烈促进了SAK的产生（181毫克/升）。此外，成熟SAK的细胞外稳定性显著增强。这些数据表明wprA基因产物在分泌过程中和细胞外环境中对外源蛋白质的降解具有重要作用。
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