Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria

Microalgae such as cyanobacteria convert CO2 to compatible drop‐in fuels, such as alkanes. However, the production yield is approximately 0.05–1.0% of the dry weight of natural algae. Here, we aimed to study the role of transcriptional expression, mRNA molecular structure and culture‐dependent accum...

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Main Authors: Misato Nagao, Takato Ozaki, Hirofumi Fukuda, Yu Kanesaki, Munehiko Asayama
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:FEBS Open Bio
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Online Access:https://doi.org/10.1002/2211-5463.70009
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author Misato Nagao
Takato Ozaki
Hirofumi Fukuda
Yu Kanesaki
Munehiko Asayama
author_facet Misato Nagao
Takato Ozaki
Hirofumi Fukuda
Yu Kanesaki
Munehiko Asayama
author_sort Misato Nagao
collection DOAJ
description Microalgae such as cyanobacteria convert CO2 to compatible drop‐in fuels, such as alkanes. However, the production yield is approximately 0.05–1.0% of the dry weight of natural algae. Here, we aimed to study the role of transcriptional expression, mRNA molecular structure and culture‐dependent accumulation of alkanes from two cyanobacteria species. The transcription start sites of the alkane biosynthesis genes ado and aar were identified in the representative cyanobacteria strains Synechocystis sp. PCC 6803 and Limnothrix sp. SK1‐2‐1, which produce heptadecane and pentadecane, respectively. This characterisation revealed the potential promoters and unique mRNA structures of the ado and aar genes in these species. Transcripts from these genes were induced more in the nitrogen‐depleted BG11 (BG11‐N) culture than in the BG11 culture, although the biomass was reduced, and as such the amount of alkanes obtained per unit medium was greater for BG11 than for BG11‐N. PCC 6803 transconjugants carrying alkane biosynthesis genes from PCC 6803 or SK1‐2‐1 showed an approximately 1.8‐ to 2.3‐fold increase in heptadecane production compared to the control strain when grown on BG11 cultures without any nitrogen depletion. These results suggest that not only the enzymes ADO/AAR but also the intracellular production of fatty acyl‐ACP substrates may be important for the mass production of target alkanes.
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spelling doaj-art-cd01d79a2ff948c8a170e7c28d14ff6d2025-08-20T01:57:14ZengWileyFEBS Open Bio2211-54632025-06-0115694996210.1002/2211-5463.70009Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteriaMisato Nagao0Takato Ozaki1Hirofumi Fukuda2Yu Kanesaki3Munehiko Asayama4College of Agriculture Ibaraki University Ibaraki JapanCollege of Agriculture Ibaraki University Ibaraki JapanCollege of Agriculture Ibaraki University Ibaraki JapanResearch Institute of Green Science and Technology Shizuoka University Shizuoka JapanCollege of Agriculture Ibaraki University Ibaraki JapanMicroalgae such as cyanobacteria convert CO2 to compatible drop‐in fuels, such as alkanes. However, the production yield is approximately 0.05–1.0% of the dry weight of natural algae. Here, we aimed to study the role of transcriptional expression, mRNA molecular structure and culture‐dependent accumulation of alkanes from two cyanobacteria species. The transcription start sites of the alkane biosynthesis genes ado and aar were identified in the representative cyanobacteria strains Synechocystis sp. PCC 6803 and Limnothrix sp. SK1‐2‐1, which produce heptadecane and pentadecane, respectively. This characterisation revealed the potential promoters and unique mRNA structures of the ado and aar genes in these species. Transcripts from these genes were induced more in the nitrogen‐depleted BG11 (BG11‐N) culture than in the BG11 culture, although the biomass was reduced, and as such the amount of alkanes obtained per unit medium was greater for BG11 than for BG11‐N. PCC 6803 transconjugants carrying alkane biosynthesis genes from PCC 6803 or SK1‐2‐1 showed an approximately 1.8‐ to 2.3‐fold increase in heptadecane production compared to the control strain when grown on BG11 cultures without any nitrogen depletion. These results suggest that not only the enzymes ADO/AAR but also the intracellular production of fatty acyl‐ACP substrates may be important for the mass production of target alkanes.https://doi.org/10.1002/2211-5463.70009alkanecyanobacteriahydrocarbon fuelprimer extensionpromotertranscription
spellingShingle Misato Nagao
Takato Ozaki
Hirofumi Fukuda
Yu Kanesaki
Munehiko Asayama
Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
FEBS Open Bio
alkane
cyanobacteria
hydrocarbon fuel
primer extension
promoter
transcription
title Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
title_full Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
title_fullStr Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
title_full_unstemmed Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
title_short Alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
title_sort alkane biosynthesis gene expression and its increased production in recombinant cyanobacteria
topic alkane
cyanobacteria
hydrocarbon fuel
primer extension
promoter
transcription
url https://doi.org/10.1002/2211-5463.70009
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AT takatoozaki alkanebiosynthesisgeneexpressionanditsincreasedproductioninrecombinantcyanobacteria
AT hirofumifukuda alkanebiosynthesisgeneexpressionanditsincreasedproductioninrecombinantcyanobacteria
AT yukanesaki alkanebiosynthesisgeneexpressionanditsincreasedproductioninrecombinantcyanobacteria
AT munehikoasayama alkanebiosynthesisgeneexpressionanditsincreasedproductioninrecombinantcyanobacteria