Abstract
Tk1884, an open reading frame encoding α-amylase in Thermococcus kodakarensis, was cloned with the native signal sequence and expressed in Escherichia coli. Heterologous gene expression resulted in secretion of the recombinant protein to the extracellular culture medium. Extracellular α-amylase activity gradually increased after induction. Tk1884 was purified from the extracellular medium, and its molecular mass determined by electrospray ionization mass spectrometry indicated the cleavage of a few amino acids. The N-terminal amino acid sequence of the purified Tk1884 was determined, which revealed that the signal peptide was cleaved between Ala26 and Ala27 by E. coli signal peptidase. To the best of our knowledge, this is the first report describing an archaeal signal sequence recognized and cleaved by E. coli signal peptidase.
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Published in Russian in Biokhimiya, 2017, Vol. 82, No. 7, pp. 1069–1074.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM17-112, May 8, 2017.
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Majida Atta Muhammad, Samia Falak, Naeem Rashid, Qurra-tul-Ann Afza Gardner, Nasir Ahmad, Tadayuki Imanaka, and Muhammad Akhtar, Escherichia coli Signal Peptidase Recognizes and Cleaves Archaeal Signal Sequence (ISSN 0006-2979, Biochemistry (Moscow), 2017, Vol. 82, No. 7, pp. 821–825)
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Muhammad, M.A., Falak, S., Rashid, N. et al. Escherichia coli signal peptidase recognizes and cleaves archaeal signal sequence. Biochemistry Moscow 82, 821–825 (2017). https://doi.org/10.1134/S0006297917070070
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DOI: https://doi.org/10.1134/S0006297917070070