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Simultaneous Uptake of Lignocellulose- Based Monosaccharides by Escherichia Coli
KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
KTH, Skolan för bioteknologi (BIO), Proteomik och nanobioteknologi. KTH, Centra, Science for Life Laboratory, SciLifeLab.
KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.ORCID-id: 0000-0002-6979-0069
2014 (engelsk)Inngår i: Biotechnology and Bioengineering, ISSN 0006-3592, E-ISSN 1097-0290, Vol. 111, nr 6, s. 1108-1115Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Lignocellulosic waste is a naturally abundant biomass and is therefore an attractive material to use in second generation biorefineries. Microbial growth on the monosaccharides present in hydrolyzed lignocellulose is however associated with several obstacles whereof one is the lack of simultaneous uptake of the sugars. We have studied the aerobic growth of Escherichia coli on D-glucose, D-xylose, and L-arabinose and for simultaneous uptake to occur, both the carbon catabolite repression mechanism (CCR) and the AraC repression of xylose uptake and metabolism had to be removed. The strain AF1000 is a MC4100 derivative that is only able to assimilate arabinose after a considerable lag phase, which is unsuitable for commercial production. This strain was successfully adapted to growth on L-arabinose and this led to simultaneous uptake of arabinose and xylose in a diauxic growth mode following glucose consumption. In this strain, a deletion in the phosphoenolpyruvate:phosphotransferase system (PTS) for glucose uptake, the ptsG mutation, was introduced. The resulting strain, PPA652ara simultaneously consumed all three monosaccharides at a maximum specific growth rate of 0.59h(-1), 55% higher than for the ptsG mutant alone. Also, no residual sugar was present in the cultivation medium. The potential of PPA652ara is further acknowledged by the performance of AF1000 during fed-batch processing on a mixture of D-glucose, D-xylose, and L-arabinose. The conclusion is that without the removal of both layers of carbon uptake control, this process results in accumulation of pentoses and leads to a reduction of the specific growth rate by 30%.

sted, utgiver, år, opplag, sider
2014. Vol. 111, nr 6, s. 1108-1115
Emneord [en]
Escherichia coli, simultaneous uptake, lignocellulose, ptsG, carbon catabolite repression
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-146542DOI: 10.1002/bit.25182ISI: 000335154300007Scopus ID: 2-s2.0-84899632399OAI: oai:DiVA.org:kth-146542DiVA, id: diva2:724117
Forskningsfinansiär
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Merknad

QC 20140612

Tilgjengelig fra: 2014-06-12 Laget: 2014-06-12 Sist oppdatert: 2017-12-05bibliografisk kontrollert
Inngår i avhandling
1. Strategies for improved Escherichia coli bioprocessing performance
Åpne denne publikasjonen i ny fane eller vindu >>Strategies for improved Escherichia coli bioprocessing performance
2015 (engelsk)Doktoravhandling, med artikler (Annet vitenskapelig)
Abstract [en]

Escherichia coli has a proven track record for successful production of anything from small molecules like organic acids to large therapeutic proteins, and has thus important applications in both R&D and commercial production. The versatility of this organism in combination with the accumulated knowledge of its genome, metabolism and physiology, has allowed for development of specialty strains capable of performing very specific tasks, opening up opportunities within new areas. The work of this thesis has been devoted to alter membrane transport proteins and the regulation of these, in order for E. coli to find further application within two such important areas.

The first area was vaccine development, where it was investigated if E. coli could be a natural vehicle for live vaccine production. The hypothesis was that the introduction and manipulation of a protein surface translocation system from pathogenic E. coli would result in stable expression levels of Salmonella subunit antigens on the surface of laboratory E. coli. While different antigen combinations were successfully expressed on the surface of E. coli, larger proteins were affected by proteolysis, which manipulation of cultivation conditions could reduce, but not eliminate completely. The surface expressed antigens were further capable of inducing proinflammatory responses in epithelial cells.

The second area was biorefining. By altering the regulation of sugar assimilation, it was hypothesized that simultaneous uptake of the sugars present in lignocellulose hydrolyzates could be achieved, thereby improving the yield and productivity of important bio-based chemicals. The dual-layered catabolite repression was identified and successfully removed in the engineered E. coli, and the compound (R)-3-hydroxybutyric acid was produced from simultaneous assimilation of glucose, xylose and arabinose.

sted, utgiver, år, opplag, sider
Stockholm: KTH Royal Institute of Technology, 2015. s. 104
Serie
TRITA-BIO-Report, ISSN 1654-2312 ; 2015:9
Emneord
E. coli, Salmonella, surface expression, autotransport, AIDA-I, lignocellulose, glucose, xylose, arabinose, simultaneous uptake, 3HB
HSV kategori
Forskningsprogram
Bioteknologi
Identifikatorer
urn:nbn:se:kth:diva-166387 (URN)978-91-7595-523-0 (ISBN)
Disputas
2015-06-05, FB52, AlbaNova universitetscentrum, Roslagstullsbacken 21, KTH, Stockholm, 10:00 (engelsk)
Opponent
Veileder
Merknad

QC 20150508

Tilgjengelig fra: 2015-05-08 Laget: 2015-05-08 Sist oppdatert: 2015-05-08bibliografisk kontrollert

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