Abstract
Streptomyces is an attractive host for heterologous protein secretion. To further optimize its expression capacity, better expression vectors will be helpful. Here, based on pSGL1, a high copy number plasmid present in Streptomyces globisporus C-1027, we constructed a series of novel E. coli-Streptomyces shuttle expression vectors pIMB2–4. These vectors, which are compatible with pIJ-derived vectors, contain the strong promoter ermE*p and signal sequence SPMelC1 of the first ORF of melanin operon in S. antibiotics (pIMB2), SPCagA of C-1027 apoprotein in S. globisporus C-1027 (pIMB3 and pIMB4). Using these vectors, human interleukin-6 (IL-6) could successfully be expressed and secreted using S. lividans TK24 as host. Furthermore, replacement of a rare leucine codon TTA with CTG in SPCagA enhanced IL-6 production.
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Acknowledgments
We thank Dr. Kenneth J. McDowall for providing plasmid pL646. We also appreciate Prof. Yuan Li and Rong Jiang for their kind help for HPLC analysis. We are especially grateful to Prof. Jozef Anné for his valuable comments and careful language revision. This work was supported by the China Ministry of Science and Technology (2006BAF07B01 and 2009BAK61B04) and the Key New Drug Creation and Manufacturing Program (2009ZX09501-008). Support is also acknowledged from the National Natural Science Foundation of China (30973668), Beijing Natural Science Foundation (5102032) and China Ministry of Education (NCET-06-0157).
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Yuanjun Zhu and Lifei Wang equally contributed to this study.
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Zhu, Y., Wang, L., Du, Y. et al. Heterologous expression of human interleukin-6 in Streptomyces lividans TK24 using novel secretory expression vectors. Biotechnol Lett 33, 253–261 (2011). https://doi.org/10.1007/s10529-010-0428-0
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DOI: https://doi.org/10.1007/s10529-010-0428-0