Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium

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Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium. / Jacobsen, Jacob Hedemand; Frigaard, Niels-Ulrik.

In: Metabolic Engineering, Vol. 21, 2014, p. 60-70.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Jacobsen, JH & Frigaard, N-U 2014, 'Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium', Metabolic Engineering, vol. 21, pp. 60-70. https://doi.org/10.1016/j.ymben.2013.11.004

APA

Jacobsen, J. H., & Frigaard, N-U. (2014). Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium. Metabolic Engineering, 21, 60-70. https://doi.org/10.1016/j.ymben.2013.11.004

Vancouver

Jacobsen JH, Frigaard N-U. Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium. Metabolic Engineering. 2014;21:60-70. https://doi.org/10.1016/j.ymben.2013.11.004

Author

Jacobsen, Jacob Hedemand ; Frigaard, Niels-Ulrik. / Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium. In: Metabolic Engineering. 2014 ; Vol. 21. pp. 60-70.

Bibtex

@article{1e83016f45404e07aa63dac6bf83d3de,
title = "Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium",
abstract = "d-Mannitol (hereafter denoted mannitol) is used in the medical and food industry and is currently produced commercially by chemical hydrogenation of fructose or by extraction from seaweed. Here, the marine cyanobacterium Synechococcus sp. PCC 7002 was genetically modified to photosynthetically produce mannitol from CO2 as the sole carbon source. Two codon-optimized genes, mannitol-1-phosphate dehydrogenase (mtlD) from Escherichia coli and mannitol-1-phosphatase (mlp) from the protozoan chicken parasite Eimeria tenella, in combination encoding a biosynthetic pathway from fructose-6-phosphate to mannitol, were expressed in the cyanobacterium resulting in accumulation of mannitol in the cells and in the culture medium. The mannitol biosynthetic genes were expressed from a single synthetic operon inserted into the cyanobacterial chromosome by homologous recombination. The mannitol biosynthesis operon was constructed using a novel uracil-specific excision reagent (USER)-based polycistronic expression system characterized by ligase-independent, directional cloning of the protein-encoding genes such that the insertion site was regenerated after each cloning step. Genetic inactivation of glycogen biosynthesis increased the yield of mannitol presumably by redirecting the metabolic flux to mannitol under conditions where glycogen normally accumulates. A total mannitol yield equivalent to 10% of cell dry weight was obtained in cell cultures synthesizing glycogen while the yield increased to 32% of cell dry weight in cell cultures deficient in glycogen synthesis; in both cases about 75% of the mannitol was released from the cells into the culture medium by an unknown mechanism. The highest productivity was obtained in a glycogen synthase deficient culture that after 12 days showed a mannitol concentration of 1.1gmannitolL(-1) and a production rate of 0.15gmannitolL(-1)day(-1). This system may be useful for biosynthesis of valuable sugars and sugar derivatives from CO2 in cyanobacteria.",
author = "Jacobsen, {Jacob Hedemand} and Niels-Ulrik Frigaard",
note = "{\textcopyright} 2013 International Metabolic Engineering Society Published by International Metabolic Engineering Society All rights reserved.",
year = "2014",
doi = "10.1016/j.ymben.2013.11.004",
language = "English",
volume = "21",
pages = "60--70",
journal = "Metabolic Engineering",
issn = "1096-7176",
publisher = "Academic Press",

}

RIS

TY - JOUR

T1 - Engineering of photosynthetic mannitol biosynthesis from CO2 in a cyanobacterium

AU - Jacobsen, Jacob Hedemand

AU - Frigaard, Niels-Ulrik

N1 - © 2013 International Metabolic Engineering Society Published by International Metabolic Engineering Society All rights reserved.

PY - 2014

Y1 - 2014

N2 - d-Mannitol (hereafter denoted mannitol) is used in the medical and food industry and is currently produced commercially by chemical hydrogenation of fructose or by extraction from seaweed. Here, the marine cyanobacterium Synechococcus sp. PCC 7002 was genetically modified to photosynthetically produce mannitol from CO2 as the sole carbon source. Two codon-optimized genes, mannitol-1-phosphate dehydrogenase (mtlD) from Escherichia coli and mannitol-1-phosphatase (mlp) from the protozoan chicken parasite Eimeria tenella, in combination encoding a biosynthetic pathway from fructose-6-phosphate to mannitol, were expressed in the cyanobacterium resulting in accumulation of mannitol in the cells and in the culture medium. The mannitol biosynthetic genes were expressed from a single synthetic operon inserted into the cyanobacterial chromosome by homologous recombination. The mannitol biosynthesis operon was constructed using a novel uracil-specific excision reagent (USER)-based polycistronic expression system characterized by ligase-independent, directional cloning of the protein-encoding genes such that the insertion site was regenerated after each cloning step. Genetic inactivation of glycogen biosynthesis increased the yield of mannitol presumably by redirecting the metabolic flux to mannitol under conditions where glycogen normally accumulates. A total mannitol yield equivalent to 10% of cell dry weight was obtained in cell cultures synthesizing glycogen while the yield increased to 32% of cell dry weight in cell cultures deficient in glycogen synthesis; in both cases about 75% of the mannitol was released from the cells into the culture medium by an unknown mechanism. The highest productivity was obtained in a glycogen synthase deficient culture that after 12 days showed a mannitol concentration of 1.1gmannitolL(-1) and a production rate of 0.15gmannitolL(-1)day(-1). This system may be useful for biosynthesis of valuable sugars and sugar derivatives from CO2 in cyanobacteria.

AB - d-Mannitol (hereafter denoted mannitol) is used in the medical and food industry and is currently produced commercially by chemical hydrogenation of fructose or by extraction from seaweed. Here, the marine cyanobacterium Synechococcus sp. PCC 7002 was genetically modified to photosynthetically produce mannitol from CO2 as the sole carbon source. Two codon-optimized genes, mannitol-1-phosphate dehydrogenase (mtlD) from Escherichia coli and mannitol-1-phosphatase (mlp) from the protozoan chicken parasite Eimeria tenella, in combination encoding a biosynthetic pathway from fructose-6-phosphate to mannitol, were expressed in the cyanobacterium resulting in accumulation of mannitol in the cells and in the culture medium. The mannitol biosynthetic genes were expressed from a single synthetic operon inserted into the cyanobacterial chromosome by homologous recombination. The mannitol biosynthesis operon was constructed using a novel uracil-specific excision reagent (USER)-based polycistronic expression system characterized by ligase-independent, directional cloning of the protein-encoding genes such that the insertion site was regenerated after each cloning step. Genetic inactivation of glycogen biosynthesis increased the yield of mannitol presumably by redirecting the metabolic flux to mannitol under conditions where glycogen normally accumulates. A total mannitol yield equivalent to 10% of cell dry weight was obtained in cell cultures synthesizing glycogen while the yield increased to 32% of cell dry weight in cell cultures deficient in glycogen synthesis; in both cases about 75% of the mannitol was released from the cells into the culture medium by an unknown mechanism. The highest productivity was obtained in a glycogen synthase deficient culture that after 12 days showed a mannitol concentration of 1.1gmannitolL(-1) and a production rate of 0.15gmannitolL(-1)day(-1). This system may be useful for biosynthesis of valuable sugars and sugar derivatives from CO2 in cyanobacteria.

U2 - 10.1016/j.ymben.2013.11.004

DO - 10.1016/j.ymben.2013.11.004

M3 - Journal article

C2 - 24269997

VL - 21

SP - 60

EP - 70

JO - Metabolic Engineering

JF - Metabolic Engineering

SN - 1096-7176

ER -

ID: 90920713