PMID- 32561588
OWN - NLM
STAT- MEDLINE
DCOM- 20201120
LR  - 20201120
IS  - 1098-5336 (Electronic)
IS  - 0099-2240 (Print)
IS  - 0099-2240 (Linking)
VI  - 86
IP  - 17
DP  - 2020 Aug 18
TI  - Applying Statistical Design of Experiments To Understanding the Effect of Growth 
      Medium Components on Cupriavidus necator H16 Growth.
LID - 10.1128/AEM.00705-20 [doi]
LID - e00705-20
AB  - Cupriavidus necator H16 is gaining significant attention as a microbial chassis 
      for range of biotechnological applications. While the bacterium is a major 
      producer of bioplastics, its lithoautotrophic and versatile metabolic 
      capabilities make the bacterium a promising microbial chassis for biofuels and 
      chemicals using renewable resources. It remains necessary to develop appropriate 
      experimental resources to permit controlled bioengineering and system 
      optimization of this microbe. In this study, we employed statistical design of 
      experiments to gain understanding of the impact of components of defined media on 
      C. necator growth and built a model that can predict the bacterium's cell density 
      based on medium components. This highlighted medium components, and interaction 
      between components, having the most effect on growth: fructose, amino acids, 
      trace elements, CaCl(2), and Na(2)HPO(4) contributed significantly to growth (t 
      values of <-1.65 or >1.65); copper and histidine were found to interact and must 
      be balanced for robust growth. Our model was experimentally validated and found 
      to correlate well (r(2) = 0.85). Model validation at large culture scales showed 
      correlations between our model-predicted growth ranks and experimentally 
      determined ranks at 100 ml in shake flasks (rho = 0.87) and 1 liter in a bioreactor 
      (rho = 0.90). Our approach provides valuable and quantifiable insights on the 
      impact of medium components on cell growth and can be applied to model other C. 
      necator responses that are crucial for its deployment as a microbial chassis. 
      This approach can be extended to other nonmodel microbes of medical and 
      industrial biotechnological importance.IMPORTANCE Chemically defined media (CDM) 
      for cultivation of C. necator vary in components and compositions. This lack of 
      consensus makes it difficult to optimize new processes for the bacterium. This 
      study employed statistical design of experiments (DOE) to understand how basic 
      components of defined media affect C. necator growth. Our growth model predicts 
      that C. necator can be cultivated to high cell density with components held at 
      low concentrations, arguing that CDM for large-scale cultivation of the bacterium 
      for industrial purposes will be economically competitive. Although existing CDM 
      for the bacterium are without amino acids, addition of a few amino acids to 
      growth medium shortened lag phase of growth. The interactions highlighted by our 
      growth model show how factors can interact with each other during a process to 
      positively or negatively affect process output. This approach is efficient, 
      relying on few well-structured experimental runs to gain maximum information on a 
      biological process, growth.
CI  - Copyright (c) 2020 Azubuike et al.
FAU - Azubuike, Christopher C
AU  - Azubuike CC
AUID- ORCID: 0000-0002-2376-3057
AD  - School of Natural and Environmental Sciences, Faculty of Science, Agriculture and 
      Engineering, Newcastle University, Newcastle-upon-Tyne, United Kingdom.
AD  - Department of Microbiology, Faculty of Science, University of Port Harcourt, Port 
      Harcourt, Rivers State, Nigeria.
FAU - Edwards, Martin G
AU  - Edwards MG
AUID- ORCID: 0000-0002-4863-653X
AD  - School of Natural and Environmental Sciences, Faculty of Science, Agriculture and 
      Engineering, Newcastle University, Newcastle-upon-Tyne, United Kingdom.
FAU - Gatehouse, Angharad M R
AU  - Gatehouse AMR
AUID- ORCID: 0000-0002-6406-2927
AD  - School of Natural and Environmental Sciences, Faculty of Science, Agriculture and 
      Engineering, Newcastle University, Newcastle-upon-Tyne, United Kingdom.
FAU - Howard, Thomas P
AU  - Howard TP
AUID- ORCID: 0000-0002-5546-4043
AD  - School of Natural and Environmental Sciences, Faculty of Science, Agriculture and 
      Engineering, Newcastle University, Newcastle-upon-Tyne, United Kingdom 
      Thomas.Howard@newcastle.ac.uk.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200818
PL  - United States
TA  - Appl Environ Microbiol
JT  - Applied and environmental microbiology
JID - 7605801
RN  - 0 (Culture Media)
SB  - IM
MH  - Culture Media/chemistry/*metabolism
MH  - Cupriavidus necator/*growth & development/metabolism
MH  - Models, Statistical
PMC - PMC7440812
OTO - NOTNLM
OT  - Cupriavidus necator H16
OT  - automation
OT  - chemically defined media
OT  - design of experiments (DOE)
EDAT- 2020/06/21 06:00
MHDA- 2020/11/21 06:00
PMCR- 2020/08/18
CRDT- 2020/06/21 06:00
PHST- 2020/03/24 00:00 [received]
PHST- 2020/05/31 00:00 [accepted]
PHST- 2020/06/21 06:00 [pubmed]
PHST- 2020/11/21 06:00 [medline]
PHST- 2020/06/21 06:00 [entrez]
PHST- 2020/08/18 00:00 [pmc-release]
AID - AEM.00705-20 [pii]
AID - 00705-20 [pii]
AID - 10.1128/AEM.00705-20 [doi]
PST - epublish
SO  - Appl Environ Microbiol. 2020 Aug 18;86(17):e00705-20. doi: 10.1128/AEM.00705-20. 
      Print 2020 Aug 18.