Biochemical characterization of polyunsaturated fatty acid synthesis in Schizochytrium: Release of the products as free fatty acids

Metz, James G.; Kuner, Jerry; Rosenzweig, Bradley; Lippmeier, J. Casey; Roessler, Paul G.; Zirkle, Ross

doi:10.1016/j.plaphy.2009.02.002

Cited by 72 publications

(62 citation statements)

References 23 publications

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“…The fi rst, which is found in several marine bacteria, is the polyketide synthase-like pathway (the PUFA synthase pathway), comprising reiterative cycles including condensation, reduction, dehydration, and isomerization steps, with each step catalyzed by different enzymes. Three functional ORFs of the PUFA synthase pathway have been identifi ed in Schizochytrium (now reassigned to Aurantiochytrium ) ( 9,20,21 ). Lippmeier et al (10) suggested that the PUFA synthase pathway is the sole system responsible for PUFA production in Schizochytrium because the disruption of an ORF of a PUFA synthase led to the loss of PUFAs in the thraustochytrids, which became PUFA-dependent auxotrophs.…”

Section: Discussionmentioning

confidence: 99%

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Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304

Matsuda

Sakaguchi

Hamaguchi

et al. 2012

Journal of Lipid Research

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Section: Discussionmentioning

confidence: 99%

“…This result indicates that Tau ⌬ 12des also accepts odd-chain fatty acids as substrates. Chang et al the synthetic machinery as free fatty acids ( 20 ). In contrast, the membrane desaturases accept a wide range of acyl substrates ( 24,25 ).…”

Section: Discussionmentioning

confidence: 99%

Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304

Matsuda

Sakaguchi

Hamaguchi

et al. 2012

Journal of Lipid Research

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“…It has been shown that PUFA producers, such as Schizochytrium species, accumulate products as free fatty acids, which are presumably released by the activity of a TE enzyme (15). However, a gene or DNA sequence encoding a dedicated TE enzyme has not been identified near any of the PUFA synthase gene clusters from deep-sea organisms.…”

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confidence: 99%

Structure, Activity, and Substrate Selectivity of the Orf6 Thioesterase from Photobacterium profundum

Rodriguez-Guilbe

Oyola‐Robles

Schreiter

et al. 2013

Journal of Biological Chemistry

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Background: Thioesterases are required for the release of polyunsaturated fatty acids in some bacteria. Results: The Orf6 protein from Photobacterium profundum has been characterized both functionally and structurally. Conclusion: Orf6 has a substrate preference for long-chain fatty acids. Significance: This is the first in vitro and structural characterization of a hotdog thioesterase from a deep-sea bacterium.

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“…It is worth noting that the scope of all reports describing the molecular tools for thraustochytrids is limited to the genus Aurantiochytrium (formerly assigned to Schizochytrium) (6,12,17). Thus, the development of a genetic system for multiple transgene expression and targeted gene knockout in thraustochytrids is urgently required for mass and/or tailor-made production of beneficial PUFAs as well as FA/ squalene-based fuels.…”

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confidence: 99%

Versatile Transformation System That Is Applicable to both Multiple Transgene Expression and Gene Targeting for Thraustochytrids

Sakaguchi

Matsuda

Kobayashi

et al. 2012

Appl Environ Microbiol

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A versatile transformation system for thraustochytrids, a promising producer for polyunsaturated fatty acids and fatty acid-derived fuels, was established. G418, hygromycin B, blasticidin, and zeocin inhibited the growth of thraustochytrids, indicating that multiple selectable marker genes could be used in the transformation system. A neomycin resistance gene (neo r ), driven with an ubiquitin or an EF-1␣ promoter-terminator from Thraustochytrium aureum ATCC 34304, was introduced into representatives of two thraustochytrid genera, Aurantiochytrium and Thraustochytrium. The neo r marker was integrated into the chromosomal DNA by random recombination and then functionally translated into neo r mRNA. Additionally, we confirmed that another two genera, Parietichytrium and Schizochytrium, could be transformed by the same method. By this method, the enhanced green fluorescent protein was functionally expressed in thraustochytrids. Meanwhile, T. aureum ATCC 34304 could be transformed by two 18S ribosomal DNA-targeting vectors, designed to cause single-or double-crossover homologous recombination. Finally, the fatty acid ⌬5 desaturase gene was disrupted by double-crossover homologous recombination in T. aureum ATCC 34304, resulting in an increase of dihomo-␥-linolenic acid (C 20:3n-6 ) and eicosatetraenoic acid (C 20:4n-3 ), substrates for ⌬5 desaturase, and a decrease of arachidonic acid (C 20:4n-6 ) and eicosapentaenoic acid (C 20:5n-3 ), products for the enzyme. These results clearly indicate that a versatile transformation system which could be applicable to both multiple transgene expression and gene targeting was established for thraustochytrids.

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Biochemical characterization of polyunsaturated fatty acid synthesis in Schizochytrium: Release of the products as free fatty acids

Cited by 72 publications

References 23 publications

Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304

Analysis of Δ12-fatty acid desaturase function revealed that two distinct pathways are active for the synthesis of PUFAs in T. aureum ATCC 34304

Structure, Activity, and Substrate Selectivity of the Orf6 Thioesterase from Photobacterium profundum

Versatile Transformation System That Is Applicable to both Multiple Transgene Expression and Gene Targeting for Thraustochytrids

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