Factors affecting the synthesis of the n-terminal methionine-free molecule of recombinant human interleukin-2 by Escherichia coli

Journal of Fermentation Technology Pub Date : 1988-01-01 DOI:10.1016/0385-6380(88)90045-3

Shigeru Fujimoto, Masanori Nakatsu, Koichi Kato, Kazuaki Kitano

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引用次数: 7

Abstract

Escherichia coli harboring the gene encoding human interleukin-2 (IL-2) produces a mixture of two recombinant IL-2 species: one with the amino-terminal alanine (rIL-2) and the other having an additional methionine residue at the amino terminus (Met-rIL-2). Ways to increase the amount of rIL-2 and its proportion to the total IL-2 were tried. Among E. coli K-12 derivatives, N4830 was an effective producer of recombinant IL-2. The production of the mixture was greatly increased by optimizing the medium ingredients or culture conditions. However, the percentage of rIL-2 in the product decreased almost linearly with an increase of the total production of recombinant IL-2 and was less than 10% under optimal culture conditions. By adding 4.1 × 10⁻⁵ M maganese and 7.4 × 10⁻⁵ M ferric ions to the medium, we succeeded in raising the percentage of rIL-2 to 50% without any decrease of the total production.

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影响大肠杆菌合成重组人白细胞介素-2 n端无蛋氨酸分子的因素

携带人类白细胞介素-2 (IL-2)编码基因的大肠杆菌产生两种重组IL-2的混合物:一种具有氨基端丙氨酸(IL-2)，另一种具有氨基端额外的蛋氨酸残基(met - IL-2)。尝试增加IL-2的量及其占总IL-2的比例。在大肠杆菌K-12衍生物中，N4830是重组IL-2的有效生产者。通过对培养基成分或培养条件的优化，可大大提高混合物的产量。然而，随着重组IL-2总产量的增加，产物中IL-2的百分比几乎呈线性下降，在最佳培养条件下，IL-2的百分比小于10%。通过在培养基中加入4.1 × 10−5 M锰离子和7.4 × 10−5 M铁离子，我们成功地将il -2的百分比提高到50%，而总产量没有下降。

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