Production of the short peptide surfactant DAMP4 from glucose or sucrose in high cell density cultures of Escherichia coli BL21(DE3)

Bruschi, Michele, Kroemer, Jens O., Steen, Jennifer A. and Nielsen, Lars K. (2014) Production of the short peptide surfactant DAMP4 from glucose or sucrose in high cell density cultures of Escherichia coli BL21(DE3). Microbial Cell Factories, 13 1: 1-9. doi:10.1186/s12934-014-0099-y


Author Bruschi, Michele
Kroemer, Jens O.
Steen, Jennifer A.
Nielsen, Lars K.
Title Production of the short peptide surfactant DAMP4 from glucose or sucrose in high cell density cultures of Escherichia coli BL21(DE3)
Formatted title
Production of the short peptide surfactant DAMP4 from glucose or sucrose in high cell density cultures of Escherichia coli BL21(DE3)
Journal name Microbial Cell Factories   Check publisher's open access policy
ISSN 1475-2859
Publication date 2014-08-19
Year available 2014
Sub-type Article (original research)
DOI 10.1186/s12934-014-0099-y
Open Access Status DOI
Volume 13
Issue 1
Start page 1
End page 9
Total pages 9
Place of publication London, United Kingdom
Publisher BioMed Central
Collection year 2015
Language eng
Formatted abstract
Background

Peptides are increasingly used in industry as highly functional materials. Bacterial production of recombinant peptides has the potential to provide large amounts of renewable and low cost peptides, however, achieving high product titers from Chemically Defined Media (CDM) supplemented with simple sugars remains challenging.

Results

In this work, the short peptide surfactant, DAMP4, was used as a model peptide to investigate production in Escherichia coli BL21(DE3), a classical strain used for protein production. Under the same fermentation conditions, switching production of DAMP4 from rich complex media to CDM resulted in a reduction in yield that could be attributed to the reduction in final cell density more so than a significant reduction in specific productivity. To maximize product titer, cell density at induction was maximized using a fed-batch approach. In fed-batch DAMP4 product titer increased 9-fold compared to batch, while maintaining 60% specific productivity. Under the fed-batch conditions, the final product titer of DAMP4 reached more than 7 g/L which is the highest titer of DAMP4 reported to date. To investigate production from sucrose, sucrose metabolism was engineered into BL21(DE3) using a simple plasmid approach. Using this strain, growth and DAMP4 production characteristics obtained from CDM supplemented with sucrose were similar to those obtained when culturing the parent strain on CDM supplemented with glucose.

Conclusions

Production of a model peptide was increased to several grams per liter using a CDM medium with either glucose or sucrose feedstock. It is hoped that this work will contribute cost reduction for production of designer peptide surfactants to facilitate their commercial application.
Keyword Peptide production
E. coli
Glucose
Sucrose
DAMP4
Recombinant proteins
Metabolic efficiency
Batch-production
E.-Coli
Expression
Biosurfactant
Purification
Foam
Construction
Cultivation
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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