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Characterization of cyanobacterial cells synthesizing 10-methyl stearic acid

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Abstract

Recently, microalgae have attracted attention as sources of biomass energy. However, fatty acids from the microalgae are mainly unsaturated and show low stability in oxygenated environments, due to oxidation of the double bonds. The branched-chain fatty acid, 10-methyl stearic acid, is synthesized from oleic acid in certain bacteria; the fatty acid is saturated, but melting point is low. Thus, it is stable in the presence of oxygen and is highly fluid. We previously demonstrated that BfaA and BfaB in Mycobacterium chlorophenolicum are involved in the synthesis of 10-methyl stearic acid from oleic acid. In this study, as a consequence of the introduction of bfaA and bfaB into the cyanobacterium, Synechocystis sp. PCC 6803, we succeeded in producing 10-methyl stearic acid, with yields up to 4.1% of the total fatty acid content. The synthesis of 10-methyl stearic acid in Synechocystis cells did not show a significant effect on photosynthetic activity, but the growth of the cells was retarded at 34 °C. We observed that the synthesis of 10-methylene stearic acid, a precursor of 10-methyl stearic acid, had an inhibitory effect on the growth of the transformants, which was mitigated under microoxic conditions. Eventually, the amount of 10-methyl stearic acid present in the sulfoquinovosyl diacylglycerol and phosphatidylglycerol of the transformants was remarkably higher than that in the monogalactosyldiacylglycerol and digalactosyldiacylglycerol. Overall, we successfully synthesized 10-methyl stearic acid in the phototroph, Synechocystis, demonstrating that it is possible to synthesize unique modified fatty acids via photosynthesis that are not naturally produced in photosynthetic organisms.

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Abbreviations

MGDG:

Monogalactosyldiacylglycerol

DGDG:

Digalactosyldiacylglycerol

SQDG:

Sulfoquinovosyldiacylglycerol

PG:

Phosphatidylglycerol

cobfaAB :

Codon-optimized bfaA and bfaB

GC:

Gas chromatography

FAME:

Fatty acid methyl ester

16:0:

Palmitic acid

16:1Δ9:

Palmitoleic acid

18:0:

Stearic acid

18:1Δ9:

Oleic acid

18:2Δ9,12:

Linoleic acid

18:3Δ6,9,12:

γ-Linolenic acid

18:3Δ9,12,15:

α-Linolenic acid

18:4Δ6,9,12,15:

Stearidonic acid

19:0Me10:

10-Methyl stearic acid

19:1∆Me10:

10-Methylene stearic acid.

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Acknowledgements

The genomic DNA of M. chlorophenolicum JCM 7439T was obtained from RIKEN BRC, which is a participant in the National BioResources Project of the MEXT, Japan.

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Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan

    Shuntaro Machida

  2. Food Research Institute, National Agriculture and Food Research Organization, Kannondai 2-1-12, Tsukuba, Ibaraki, 305-8642, Japan

    Shuntaro Machida

  3. Faculty of Life and Environmental Sciences, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki, 305-8572, Japan

    Iwane Suzuki

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  1. Shuntaro Machida
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  2. Iwane Suzuki
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Correspondence to Iwane Suzuki.

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Machida, S., Suzuki, I. Characterization of cyanobacterial cells synthesizing 10-methyl stearic acid. Photosynth Res 139, 173–183 (2019). https://doi.org/10.1007/s11120-018-0537-5

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