Controlled production of stable heterologous proteins in Lactococcus lactis
Controlled production of stable heterologous proteins in Lactococcus lactis
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DOI：
10.1128/aem.68.6.3141-3146.2002
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发表时间：
2002-06-01
影响因子：
4.4
通讯作者：
Langella, P
中科院分区：
文献类型：
--
作者：
Miyoshi, A;Poquet, I;Langella, P
在某些情况下，使用乳酸乳球菌（最广泛表征的乳酸菌）作为异源蛋白质的递送生物体，由于表面蛋白水解而受到生产水平低和产品质量差的限制。在这项研究中，我们在一种乳酸乳球菌菌株中结合使用乳链菌肽诱导型启动子 P-nisA 和细胞外管家蛋白酶 HtrA 的失活。通过使用葡萄球菌核酸酶 (Nuc) 和以下四种与 Nuc 融合或未融合的异源蛋白（最初在野生型乳酸乳球菌菌株中不稳定），证实了突变菌株（命名为 htrA-NZ9000）产生高水平稳定蛋白的能力：(i) 猪葡萄球菌脂肪酶，(ii) 牛轮状病毒抗原 非结构蛋白 4，(iii) 人乳头瘤病毒抗原 E7，和 (iv) 流产布鲁氏菌抗原 L7/L12。在所有情况下，菌株 htrA-NZ9000 中的蛋白质降解均显着降低，这证明了该菌株对于稳定的异源蛋白质生产的有用性。
The use of Lactococcus lactis (the most extensively characterized lactic acid bacterium) as a delivery organism for heterologous proteins is, in some cases, limited by low production levels and poor-quality products due to surface proteolysis. In this study, we combined in one L. lactis strain use of the nisin-inducible promoter P-nisA and inactivation of the extracellular housekeeping protease HtrA. The ability of the mutant strain, designated htrA-NZ9000, to produce high levels of stable proteins was confirmed by using the staphylococcal nuclease (Nuc) and the following four heterologous proteins fused or not fused to Nuc that were initially unstable in wild-type L. lactis strains: (i) Staphylococcus hyicus lipase, (ii) the bovine rotavirus antigen nonstructural protein 4, (iii) human papillomavirus antigen E7, and (iv) Brucella abortus antigen L7/L12. In all cases, protein degradation was significantly lower in strain htrA-NZ9000, demonstrating the usefulness of this strain for stable heterologous protein production.