Abstract
ɛ-Poly-L-lysine (ɛ-PL) is an L-lysine linear homopolymer, which is produced by bacteria belonging to the Streptomycetaceae family and by ergot fungi. However, the production of ɛ-PL by the wild bacteria strain is very low, which limits its utilization. In most bacteria including the Streptomyces genus, L-lysine is a precursor of ɛ-PL and is biosynthesized by the L-aspartate pathway. Aspartokinase (Ask) is the first key enzyme in this pathway and is subject to complex regulation such as the feedback inhibition by the end product amino acids. In addition, phosphoenolpyruvate carboxykinase is feedback-regulated by L-aspartate. To reduce these feedback inhibitions and to improve ɛ-PL productivity, resistant mutants were produced using sulfaguanidine (SG) + glycine + L-lysine + DL-3-hydroxynorvaline (AHV) as selective markers. Using the interaction between ɛ-PL and the charged dye in the solid culture medium, hundreds of colonies were simultaneously screened in a quick and effective manner. Finally, one ɛ-PL-producing strain, Streptomyces diastatochromogenes L9, was selected. The productivity of this strain during flask fermentation was 0.77 g/L, which was 15% higher than that of the original strain. Moreover, its fermentation performance and genetic characteristics were very stable.
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Shima S, Sakai H. Polylysine produced by Streptomyces. Agricultural and Biological Chemistry, 1977, 41(9): 1807–1809
Hiraki J, Ichikawa T, Ninomiya S, Seki H, Uohama K, Seki H, Kimura S, Yanagimoto Y, Barnett J W Jr. Use of ADME studies to confirm the safety of ɛ-polylysine as a preservative in food. Regulatory Toxicology and Pharmacology, 2003, 37(2): 328–340
Tsujita T, Takaichi H, Takaku T, Aoyama S, Hiraki J. Antiobesity action of ɛ-polylysine, a potent inhibitor of pancreatic lipase. Journal of Lipid Research, 2006, 47(8): 1852–1858
Yoshida T, Nagasawa T. ɛ-Poly-L-lysine: microbial production, biodegradation and application potential. Applied Microbiology and Biotechnology, 2003, 62(1): 21–26
Shih I L, Shen M H, Van Y T. Microbial synthesis of poly(ɛ-lysine) and its various applications. Bioresource Technology, 2006, 97(9): 1148–1159
Jun H, Masakazu H, Hiroshi M, Yoshikazu I. Improved ɛ-poly-L-Lysine production of an S-(2-aminoethyl)-L-cysteine resistant mutant of Streptomyces albulus. Seibutsu Kogakkaishi, 1998, 76: 487–493
Kahar P, Iwata T, Hiraki J, Park E Y, Okabe M. Enhancement of ɛ-polylysine production by Streptomyces albulus strain 410 using pH control. Journal of Bioscience and Bioengineering, 2001, 91(2): 190–194
Nishikawa M, Ogawa K. Distribution of microbes producing antimicrobial ɛ-poly-L-lysine polymers in soil microflora determined by a novel method. Applied and Environmental Microbiology, 2002, 68(7): 3575–3581
Hirohara H, Takehara M, Saimura M, Ikezaki A, Miyamoto M. Biosynthesis of poly(ɛ-L-lysine)s in two newly isolated strains of Streptomyces sp. Applied Microbiology and Biotechnology, 2006, 73(2): 321–331
Takehara M, Hibino A, Saimura M, Hirohara H. High-yield production of short chain length poly(ɛ-L-lysine) consisting of 5–20 residues by Streptomyces aureofaciens, and its antimicrobial activity. Biotechnology Letters, 2010, 32(9): 1299–1303
Nishikawa M, Kobayashi K. Streptomyces roseoverticillatus produces two different poly(amino acid)s: lariat-shaped gammapoly( L-glutamic acid) and epsilon-poly(L-lysine). Microbiology-Sgm, 2009, 155(Pt 9): 2988–2993
Jia S, Xu C, Tan Z, Cao W, Ou H, He X, Deng Z. Isolation characterization of a new ɛ-poly-L-lysine-producing strain TUST-2. Acta Microbiologica Sinica, 2010, 50(2): 191–196 (in Chinese)
Hamano Y, Nicchu I, Shimizu T, Onji Y, Hiraki J, Takagi H. ɛ-Poly-L-lysine producer, Streptomyces albulus, has feedback-inhibition resistant aspartokinase. Applied Microbiology and Biotechnology, 2007, 76(4): 873–882
Cao W, Tan Z, Yuan G, Jia S. Improvement of the assay method for ɛ-polylysine. Journal of Tianjin University of Science & Technology, 2007, 22: 9–11 (in Chinese)
Kawai T, Kubota T, Hiraki J, Izumi Y. Biosynthesis of ɛ-poly-Llysine in a cell-free system of Streptomyces albulus. Biochemical and Biophysical Research Communications, 2003, 311(3): 635–640
Kahar P, Kobayashi K, Iwata T, Hiraki J, Kojima M, Okabe M. Production of ɛ-polylysine in an airlift bioreactor (ABR). Journal of Bioscience and Bioengineering, 2002, 93(3): 274–280
Hirohara H, Saimura M, Takehara M, Miyamoto M, Ikezaki A. Substantially monodispersed poly(ɛ-L-lysine)s frequently occurred in newly isolated strains of Streptomyces sp. Applied Microbiology and Biotechnology, 2007, 76(5): 1009–1016
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Wang, T., Jia, S., Tan, Z. et al. Mutagenesis and selective breeding of a high producing ɛ-poly-L-lysine strain. Front. Chem. Sci. Eng. 6, 179–183 (2012). https://doi.org/10.1007/s11705-012-1273-6
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DOI: https://doi.org/10.1007/s11705-012-1273-6