Non-classical secretion of a type I L-asparaginase in Bacillus subtilis.
Non-classical secretion of a type I L-asparaginase in Bacillus subtilis.
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DOI：
10.1016/j.ijbiomac.2021.03.104
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发表时间：
2021-03
影响因子：
8.2
通讯作者：
Jia-feng Niu;Fanqiang Meng;Yawen Zhou;Chong Zhang;Zhaoxin Lu;F. Lu;Meirong Chen
中科院分区：
文献类型：
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作者：
Jia-feng Niu;Fanqiang Meng;Yawen Zhou;Chong Zhang;Zhaoxin Lu;F. Lu;Meirong Chen
L-天冬酰胺酶（EC 3.5.1.1）由于其对急性淋巴细胞白血病（ALL）、淋巴系统恶性肿瘤和霍奇金病的有效治疗以及其在防止油炸和烘焙食品中丙烯酰胺形成中的用途而显示出巨大的商业价值。本研究克隆了地衣芽孢杆菌Z-1的I型L-天冬酰胺酶基因（BlAase），并在枯草芽孢杆菌RIK 1285中表达。结果表明，即使没有任何N端信号肽的介导，BlAase也能有效地分泌到培养基中。进一步的研究表明，BlAase的分泌途径既不依赖于Sec，也不依赖于Tat，而其N端和C端区域对BlAase的表达和分泌都是必不可少的，这表明BlAase可能是通过非经典的分泌途径分泌的。为了探索其分泌能力，BlAase被用作信号肽来指导各种异源蛋白的分泌，其中五种蛋白中的两种在BlAase的介导下成功分泌。据我们所知，这是第一次在B.枯草芽孢杆菌表达的重组蛋白，为分泌表达于枯草芽孢杆菌中的重组蛋白提供了潜在的工具。枯草杆菌以BlAase为信号肽。
L-asparaginase (EC 3.5.1.1) showed great commercial value owing to its effective treatment of acute lymphoblastic leukemia (ALL), lymphoid system malignancies and Hodgkin disease, and also to its use in the prevention of acrylamide formation in fried and baked foods. In this study, a type I L-asparaginase gene fromBacillus licheniformisZ-1 (BlAase) was cloned and expressed inBacillus subtilisRIK 1285. Results showed that even without the mediation of any N-terminal signal peptides, BlAase can efficiently secrete into the medium. Further investigation indicated that the secretion of the BlAase was via neither Sec- nor Tat-dependent secretion pathway, and both the N- and C-terminal regions of the BlAase were essential for its expression and secretion, implying that BlAase might be secreted via a non-classical secretion pathway. To explore its secretion ability, BlAase was used as a signal peptide to direct the secretion of various heterologous proteins, where two of five proteins were successfully secreted with the mediation of BlAase. To the best of our knowledge, this is the first time to achieve extracellular expression of L-asparaginase via non-classical protein secretion pathway inB. subtilis, and provide a potential tool for secretion of recombinant proteins expressed inB. subtilisusing BlAase as a signal peptide.