Engineering Mannitol Biosynthesis in Escherichia
coli and Synechococcus sp. PCC 7002 Using
a Green Algal Fusion Protein
Version 2 2019-01-12, 00:43
Version 1 2018-11-21, 13:50
Posted on 2019-01-12 - 00:43
The genetic engineering of microbial
cell factories is a sustainable
alternative to the chemical synthesis of organic compounds. Successful
metabolic engineering often depends on manipulating several enzymes,
requiring multiple transformation steps and selection markers, as
well as protein assembly and efficient substrate channeling. Naturally
occurring fusion genes encoding two or more enzymatic functions may
offer an opportunity to simplify the engineering process and to generate
ready-made protein modules, but their functionality in heterologous
systems remains to be tested. Here we show that heterologous expression
of a fusion enzyme from the marine alga Micromonas pusilla, comprising a mannitol-1-phosphate dehydrogenase and a mannitol-1-phosphatase,
leads to synthesis of mannitol by Escherichia coli and by the cyanobacterium Synechococcus sp. PCC
7002. Neither of the heterologous systems naturally produce this sugar
alcohol, which is widely used in food, pharmaceutical, medical, and
chemical industries. While the mannitol production rates obtained
by single-gene manipulation were lower than those previously achieved
after pathway optimization with multiple genes, our findings show
that naturally occurring fusion proteins can offer simple building
blocks for the assembly and optimization of recombinant metabolic
pathways.
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Madsen, Mary Ann; Semerdzhiev, Stefan; Amtmann, Anna; Tonon, Thierry (2018). Engineering Mannitol Biosynthesis in Escherichia
coli and Synechococcus sp. PCC 7002 Using
a Green Algal Fusion Protein. ACS Publications. Collection. https://doi.org/10.1021/acssynbio.8b00238
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AUTHORS (4)
MM
Mary Ann Madsen
SS
Stefan Semerdzhiev
AA
Anna Amtmann
TT
Thierry Tonon