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Enhanced l-phenylalanine production by recombinant Escherichia coli BR-42 (pAP-B03) resistant to bacteriophage BP-1 via a two-stage feeding approach

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Journal of Industrial Microbiology & Biotechnology

Abstract

The l-phenylalanine (l-Phe) production by Escherichia coli WSH-Z06 (pAP-B03) was frequently prevented by bacteriophage BP-1 infestation. To cope with the bacteriophage BP-1 problem for an improved l-Phe production, one bacteriophage BP-1-resistant mutant, E. coli BR-42, was obtained from 416 mutant colonies of E. coli WSH-Z06 after N-methyl-N’-nitro-N-nitrosoguanidine (NTG) mutagenesis by selection for resistance to bacteriophage BP-1. The recombinant E. coli BR-42-carrying plasmid pAP-B03 had a high capacity in l-Phe production and a remarkable tolerance to 1 × 1010 pfu (plaque-forming unit)/ml bacteriophage stock. For an enhanced l-Phe production by E. coli BR-42 (pAP-B03), the effects of different feeding strategies including pH–stat, constant rate feeding, linear decreasing rate feeding, and exponential feeding on l-Phe production were investigated; and a two-stage feeding strategy, namely exponential feeding at μ set = 0.18 h−1 in the first 20 h and a following linear varying rate feeding with F = (−0.55 × t + 18.6) ml/h, was developed to improve l-Phe production. With this two-stage feeding approach, a maximum l-Phe titer of 57.63 g/l with a high l-Phe productivity (1.15 g/l/h) was achieved, which was 15% higher than the highest level (50 g/l) reported so far according to our knowledge. The recombinant E. coli BR-42 (pAP-B03) is a potential l-Phe over-producer in substantial prevention of bacteriophage BP-1 infestation compared to its parent strain WSH-Z06 (pAP-B03).

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Acknowledgments

Funding for this research was supported by a grant from National Science Fund for Distinguished Young Scholars (No. 20625619), the Key Program of National Natural Science Foundation of China (No. 20836003), the Key Project of Chinese National Programs for Fundamental Research and Development (973 program, No. 2007CB71403), the Program for Changjiang Scholars and Innovative Research Team in University (No. IRT0532), the Fundamental Research Funds for the Central Universities (No. JUSRP10917), the Fundamental Research Funds for the Central Universities (No. JUSRP30901), and 973 Project (2010CB535014).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, Jiangsu, China

    Haiyan Zhou, Long Liu, Tianwen Wang, Guocheng Du & Jian Chen

  2. School of Biotechnology, Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, China

    Haiyan Zhou, Long Liu, Tianwen Wang, Guocheng Du & Jian Chen

  3. School of Life Sciences, Shanghai University, Shanghai, 200444, China

    Xianyan Liao

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  1. Haiyan Zhou
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Correspondence to Tianwen Wang or Guocheng Du.

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Zhou, H., Liao, X., Liu, L. et al. Enhanced l-phenylalanine production by recombinant Escherichia coli BR-42 (pAP-B03) resistant to bacteriophage BP-1 via a two-stage feeding approach. J Ind Microbiol Biotechnol 38, 1219–1227 (2011). https://doi.org/10.1007/s10295-010-0900-9

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  • DOI: https://doi.org/10.1007/s10295-010-0900-9

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