Applied Microbiology and Biotechnology

, Volume 102, Issue 10, pp 4499–4510 | Cite as

Construction of Halomonas bluephagenesis capable of high cell density growth for efficient PHA production

  • Yilin Ren
  • Chen Ling
  • Ivan Hajnal
  • Qiong Wu
  • Guo-Qiang ChenEmail author
Biotechnological products and process engineering


High-cell-density cultivation is an effective way to improve the productivity of microbial fermentations and in turn reduce the cost of the final products, especially in the case of intracellular products. Halomonas bluephagenesis TD01 is a halophilic platform bacterium for the next generation of industrial biotechnology with a native PHA synthetic pathway, able to grow under non-sterile continuous fermentation conditions. A selection strategy for mutant strains that can grow to a high cell density was developed. Based on an error-prone DNA polymerase III ε subunit, a genome-wide random mutagenesis system was established and used in conjunction with an artificial high cell density culture environment during the selection process. A high-cell-density H. bluephagenesis TDHCD-R3 obtained after 3 rounds of selection showed an obvious enhancement of resistance to toxic metabolites including acetate, formate, lactate and ethanol compared to wild-type. H. bluephagenesis TDHCD-R3-8-3 constructed from H. bluephagenesis TDHCD-R3 by overexpressing an optimized phaCAB operon was able to grow to 15 g/L cell dry weight (CDW) containing 94% PHA in shake flask studies. H. bluephagenesis TDHCD-R3-8-3 was grown to more than 90 g/L CDW containing 79% PHA compared with only 81 g/L with 70% PHA by the wild type when incubated in a 7-L fermentor under the same conditions.


High-cell-density growth Halomonas bluephagenesis PHB Polyhydroxyalkanoates Next generation industrial biotechnology 



SEVA plasmids were kindly donated by Prof. Victor de Lorenzo from the National Centre for Biotechnology in Madrid, Spain.


This study was funded by the Ministry of Sciences and Technology of China (Grant No. 2016YFB0302504), the National Natural Science Foundation of China (Grant No. 31430003 and 31600072) and the Tsinghua President Fund (Grant No. 2015THZ10).

Compliance with ethical standards

Conflict of interest

Yilin Ren declares that he has no conflict of interest.

Chen Ling declares that he has no conflict of interest.

Ivan Hajnal declares that he has no conflict of interest.

Qiong Wu declares that he has no conflict of interest.

Guo-Qiang Chen declares that he has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Yilin Ren
    • 1
  • Chen Ling
    • 1
  • Ivan Hajnal
    • 1
  • Qiong Wu
    • 1
  • Guo-Qiang Chen
    • 1
    • 2
    • 3
    Email author
  1. 1.MOE Key Lab of Bioinformatics, Department of Biological Science and Biotechnology, School of Life Science, Tsinghua-Peking Center for Life SciencesTsinghua UniversityBeijingChina
  2. 2.Center for Nano and Micro-MechanicsTsinghua UniversityBeijingChina
  3. 3.MOE Key Lab for Industrial BiocatalysisTsinghua UniversityBeijingChina

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