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2012, Volume 6, Issue 2

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Frontiers of Chemical Science and Engineering >> 2012, Volume 6, Issue 2 doi: 10.1007/s11705-012-1273-6

Mutagenesis and selective breeding of a high producing

Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science & Technology, Tianjin 300457, China

Available online: 2012-06-05
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Abstract

-Poly- -lysine ( -PL) is an -lysine linear homopolymer, which is produced by bacteria belonging to the family and by ergot fungi. However, the production of -PL by the wild bacteria strain is very low, which limits its utilization. In most bacteria including the genus, -lysine is a precursor of -PL and is biosynthesized by the -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 -aspartate. To reduce these feedback inhibitions and to improve -PL productivity, resistant mutants were produced using sulfaguanidine (SG) + glycine+ -lysine+ -3-hydroxynorvaline (AHV) as selective markers. Using the interaction between -PL and the charged dye in the solid culture medium, hundreds of colonies were simultaneously screened in a quick and effective manner. Finally, one -PL-producing strain, 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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