Physiomics Array: A Platform for Genome Research and Cultivation of Difficult-to-Cultivate Microorganisms Final Technical Report Page: 1 of 12
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Final Report
Title: Physiomics Array: A Platform for Genome Research and
Cultivation of Difficult-to-Cultivate Microorganisms
Project ID: 0007067
Prog Mgr: Daniel W. Drell
Phone: 301-903-4742
Division: SC-72
PI: Jay D. Keasling
Award Register#: ER63144 0007067
Final Report
Executive Summary
A scalable array technology for parametric control of high-throughput cell cultivations is
demonstrated. The technology makes use of commercial printed circuit board (PCB)
technology, integrated circuit sensors, and an electrochemical gas generation system. We
present results for an array of eight 250 l microbioreactors. Each bioreactor contains an
independently addressable suite that provides closed-loop temperature control, generates
feed gas electrochemically, and continuously monitors optical density. The PCB
technology allows for the assembly of additional off-the-shelf components into the
microbioreactor array; we demonstrate the use of a commercial ISFET chip to
continuously monitor culture pH. The electrochemical dosing system provides a powerful
paradigm for reproducible gas delivery to high-density arrays of microreactors. We have
scaled the technology to a standard 96-well format and have constructed a system that
could be easily assembled.
Introduction
There is a pressing need for reliable, high-throughput cell cultivation technology. The
growing library of genomic data is expanding the number of possible modifications to
cultured species. Additionally, recent estimates indicate a large number of
microorganisms found in the environment are unculturable using current techniques or
conditions (Cowan 2000; Gao and Moore 1996; Moter and Gobel 2000; Pillai 1997).
Increasingly, the researcher will explore an extensive cultivation parameter space when
optimizing metabolic activity or bioconversions. Despite this, bioprocess experimentation
and cultivation has remained largely unchanged for decades, relying mostly on shake
flasks and 1-10 l bioreactors (Schugerl 2001; Stanbury, et al. 1995).
The miniaturization of cultivation technology has been hampered by the lack of suitably
durable, compact and relevant sensors and by the limited ability to control parameters
within small volumes (Harms, et al. 2002; Scheller, et al. 2001; Schugerl 2001; Walther,
et al. 1994). Applicable sensor technology has been progressing steadily; methods
employing microfabricated silicon-based sensors or optochemical sensing allow for the
monitoring of parameters in many simultaneous experiments (Pons 1993; Scheller, et al.
2001; Zanzotto, et al. 2002; Zhang, et al. 2002). Parameter control, however, is still
relatively primitive. For example, even the more successful miniaturized bioreactor
designs utilize tiny, hand assembled tubing to provide oxygen to the culture (Kostov, et
al. 2001). Several commercial cell culture plates (e.g. www.synthecon.com;Page 1 of 12
Keasling
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Keasling, Jay D. Physiomics Array: A Platform for Genome Research and Cultivation of Difficult-to-Cultivate Microorganisms Final Technical Report, report, July 10, 2006; United States. (https://digital.library.unt.edu/ark:/67531/metadc891975/m1/1/: accessed April 25, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.