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Vibrio natriegens as a superior host for the production of c-type cytochromes and difficult-to-express redox proteins
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Vibrio natriegens as a superior host for the production of c-type cytochromes and difficult-to-express redox proteins
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Journal Article
Vibrio natriegens as a superior host for the production of c-type cytochromes and difficult-to-express redox proteins
2024
Overview
C-type cytochromes fulfil many essential roles in both aerobic and anaerobic respiration. Their characterization requires large quantities of protein which can be obtained through heterologous production. Heterologous production of c-type cytochromes in
Escherichia coli
is hindered since the
ccmABCDEFGH
genes necessary for incorporation of heme c are only expressed under anaerobic conditions. Different strategies were devised to bypass this obstacle, such as co-expressing the
ccm
genes from the pEC86 vector. However, co-expression methods restrict the choice of expression host and vector. Here we describe the first use of
Vibrio natriegens
V
max
X2 for the recombinant production of difficult-to-express redox proteins from the extreme acidophile
Acidithiobacillus ferrooxidans
CCM4253, including three c-type cytochromes. Co-expression of the
ccm
genes was not required to produce holo-c-type cytochromes in V
max
X2.
E. coli
T7 Express only produced holo-c-type cytochromes during co-expression of the
ccm
genes and was not able to produce the inner membrane cytochrome CycA. Additionally, V
max
X2 cell extracts contained higher portions of recombinant holo-proteins than T7 Express cell extracts. All redox proteins were translocated to the intended cell compartment in both hosts. In conclusion,
V. natriegens
represents a promising alternative for the production of c-type cytochromes and difficult-to-express redox proteins.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
