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Abstract
Biopolymers such as polyhydroxybutyrate (PHB) are
considered to be carbon-neutral, and thus environmentfriendly, replacements for fossil fuel-derived plastics. They
are more expensive, however, and production process
costs must be reduced to increase market acceptance.
Alternative feedstocks offer a promising way to reduce
costs.
This application note presents the process development
Introduction
Research into polyhydroxyalkanoate (PHA) biopolymers
(including PHB) has intensified. Since their mechanical,
physical, and thermal characteristics are similar to many
fossil-fuel-based plastics, such as polypropylene (PP) and
polyethylene (PE), they have the potential to replace those
high-volume products in certain applications. Naturally
produced from sugars by various bacterial strains, they are
an environmentally friendly alternative to plastics derived
from petroleum and natural gas. PHAs are biodegradable,
non-toxic, and can either be thermoplastic or elastomeric
materials, making them suitable for applications in
biomedicine, packaging, and many other fields.
In 2014, the global production of PHAs was estimated
at 54kilotonnes, with a more than 5-fold market increase
expected by 2020 [1]. But although production costs have
decreased substantially over the last several decades, PHA
prices are still significantly higher than traditional plastics.
Feedstocks account for 50% of their average production
costs [2]. Using cheaper feedstocks, such as lignocellulosic
sugars, would be a major breakthrough in cutting costs.
Xylose [g/L]
Total [g/L]
Xylose [%]
573
270
843
32
II
532
315
847
37
III
491
360
851
42
IV
450
405
855
47
Hydr.
465
269
733
37
Figure 2: Growth and PHB formation in four 2 L fermentations (I IV, see table 1) fed with feeding solutions containing varying glucose/
xylose ratio. A: Dry cell weight (DCW) and PHB concentration; B: Glucose and xylose concentrations
Feed
Hydrolysate
DCW
[g/L]
123.4
PHB
[g/L]
63.6
PHB
[%]
51.6
YX/S
[g/g]
0.42
YP/S
[g/g]
0.23
10 L
Hydrolysate
123.0
64.6
52.5
0.40
0.19
50 L
Sugar mix
101.6
56.3
55.4
0.42
0.25
200 L
Sugar mix
129.0
71.1
56.2
0.45
0.22
Conclusion
The results confirm that the process engineers at Biotrend
achieved a successful fermentation scale up and validated
the scalability of the process. Despite the challenges of
scaling up bioprocesses, fine-tuning the fermentation
protocol in smaller scale transitions resulted in robust and
scalable processes. Scale up based on maintaining a constant
tip speed is a common strategy, but adjustments may be
necessary to manage a scalable yield. The careful, rational,
and step-by-step approach, using 5-fold scale increases
worked well. The sequence of fermentation trials at
Literature
[1] Ravenstijn J (2014): PHA... Is it here to stay? Presentation. http://www.kcpk.nl/algemeen/bijeenkomsten/presentaties/20140508jan-ravenstijn-pha-is-it-here-to-stay
[2] E4tech, RE-CORD and WUR (2015): From the Sugar Platform to biofuels and biochemicals. Final report for the European Commission, contract No. ENER/C2/423-2012/SI2.673791
[3] Kim BS, Lee SC, Lee SY, Chang HN, Chang YK, Woo SI (1994) Production of poly(3- hydroxybutyric acid) by fed-batch culture of
Alcaligenes eutrophus with glucose concentration control. Biotechnol. Bioeng. 43:8928.
[4] Lei S. (2011) Biorefining of lignocellulosic feedstock by AFEX-pretreatment and enzymatic hydrolysis for production of fermentable sugar Biorefinery.de GmbH, Hanover, Germany: German Russian Forum Biotechnology.
[5] Cesrio MT, Raposo RS, de Almeida MCMD, van Keulen F, Ferreira BS and da Fonseca MMR (2014) Enhanced bioproduction of
poly-3-hydroxybutyrate from wheat straw lignocellulosic hydrolysates. New Biotechnology 31: 104-113.
Ordering information
Description
Order no.
New Brunswick BioFlo 115, Configured Master Control Station with TMFC
New Brunswick BioFlo 115, 2 L Advanced Fermentation Vessel Kit
New Brunswick BioFlo 415
www.eppendorf.com
This work has received funding from the European Union 7th Framework Programme (FP7/20072013) under Grant Agreement number 246449 BUGWORKERS.
The authors would like to acknowledge biorefinery.de GmbH (Germany) for providing the hydrolysates.
Biotrend is a registered trademark of Biotrend Inovao e Engenharia em Biotecnologia, S.A., Portugal. Eppendorf and the Eppendorf logo are registered trademarks of
Eppendorf AG, Germany. New Brunswick is a trademark of Eppendorf AG, Germany. BioFlo and CelliGen are registered trademarks of Eppendorf, Inc., USA. U.S. Design
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