Enhanced production of polyunsaturated fatty acids by enzyme engineering of tandem acyl carrier proteins

Hayashi, Shinobu; Satoh, Yasuharu; Ujihara, Tetsuro; Takata, Yasumitsu; Dairi, Tohru

doi:10.1038/srep35441

Cited by 59 publications

(40 citation statements)

References 31 publications

(40 reference statements)

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“…Similar to previous studies, expressed sequence tag (EST) sequencing or PCR-based detection failed to identify the probable desaturases in the FAS pathway [72]. The modification of FAs is performed to produce long-chain DHA (C22:6) by an enzyme-dependent continuous process [73]. On the other side, it has been reported that marine bacteria could produce polyunsaturated fatty acid via the polyketide synthases (PKS) pathway [74].…”

Section: Pks Pathwaysupporting

confidence: 60%

Comparative Transcriptomic Analysis Uncovers Genes Responsible for the DHA Enhancement in the Mutant Aurantiochytrium sp.

Liu

et al. 2020

Microorganisms

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Docosahexaenoic acid (DHA), a n-3 long-chain polyunsaturated fatty acid, is critical for physiological activities of the human body. Marine eukaryote Aurantiochytrium sp. is considered a promising source for DHA production. Mutational studies have shown that ultraviolet (UV) irradiation (50 W, 30 s) could be utilized as a breeding strategy for obtaining high-yield DHA-producing Aurantiochytrium sp. After UV irradiation (50 W, 30 s), the mutant strain X2 which shows enhanced lipid (1.79-fold, 1417.37 mg/L) and DHA (1.90-fold, 624.93 mg/L) production, was selected from the wild Aurantiochytrium sp. Instead of eicosapentaenoic acid (EPA), 9.07% of docosapentaenoic acid (DPA) was observed in the mutant strain X2. The comparative transcriptomic analysis showed that in both wild type and mutant strain, the fatty acid synthesis (FAS) pathway was incomplete with key desaturases, but genes related to the polyketide synthase (PKS) pathway were observed. Results presented that mRNA expression levels of CoAT, AT, ER, DH, and MT down-regulated in wild type but up-regulated in mutant strain X2, corresponding to the increased intercellular DHA accumulation. These findings indicated that CoAT, AT, ER, DH, and MT can be exploited for high DHA yields in Aurantiochytrium.

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Section: Pks Pathwaysupporting

confidence: 60%

Comparative Transcriptomic Analysis Uncovers Genes Responsible for the DHA Enhancement in the Mutant Aurantiochytrium sp.

Liu

et al. 2020

Microorganisms

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“…[3] In contrast, only DHA producers have been isolated from microalgae such as Schizochytrium sp.a nd Aurantiochytrium sp. [5] Second, PUFAs ynthases possess two types of dehydratase,ap olyketide synthase (PKS) type dehydratase (DH PKS )a nd FabA-type dehydratase (DH FabA ). However,PUFAsynthases have unique features distinct from FASs.First, PUFAsynthases have multitandem ACPdomains and we previously showed that PUFAp roductivities increased, depending on the number of ACPs,without profile changes.…”

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confidence: 99%

“…GC-MS analysis traces at m/z 79[5] of products produced in the gene replacementassay.A )Gene replacementofepa genes with dha genes (from 2nd to 5th). Standards (top) and epa-ABCD (bottom).B)Gene replacementofdha genes with epa genes (from 2nd to 5th).…”

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confidence: 99%

Control Mechanism for Carbon‐Chain Length in Polyunsaturated Fatty‐Acid Synthases

et al. 2019

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“…These genes were cloned into different and compatible expression vectors and introduced into E. coli BLR(DE3)Δ fadE for evaluation of PUFA profiles. Because ARA productivity was significantly lower than that of EPA in the heterologous expression system, the PUFA profiles were evaluated by replacing each epa gene with the corresponding ara gene. When epa‐D was replaced with ara‐D , the PUFA profile was almost the same as that of the parents.…”

Section: Figurementioning

confidence: 99%

“… GC‐MS analysis trace at m / z 79 of products produced in the gene replacement assay. A) Gene replacement of epa genes with ara genes (from 3 rd to bottom).…”

Section: Figurementioning

confidence: 99%

Control Mechanism for cis Double‐Bond Formation by Polyunsaturated Fatty‐Acid Synthases

et al. 2019

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Polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) are essential fatty acids for humans. Some microorganisms biosynthesize these PUFAs through PUFA synthases composed of four subunits with multiple catalytic domains. These PUFA synthases each create a specific PUFA without undesirable byproducts, even though the multiple catalytic domains in each large subunit are very similar. However, the detailed biosynthetic pathways and mechanisms for controlling final‐product profiles are still obscure. In this study, the FabA‐type dehydratase domain (DHFabA) in the C‐subunit and the polyketide synthase‐type dehydratase domain (DHPKS) in the B‐subunit of ARA synthase were revealed to be essential for ARA biosynthesis by in vivo gene exchange assays. Furthermore, in vitro analysis with truncated recombinant enzymes and C4‐ to C8‐acyl ACP substrates showed that ARA and EPA synthases utilized two types of DH domains, DHPKS and DHFabA, depending on the carbon‐chain length, to introduce either saturation or cis double bonds to growing acyl chains.

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Enhanced production of polyunsaturated fatty acids by enzyme engineering of tandem acyl carrier proteins

Cited by 59 publications

References 31 publications

Comparative Transcriptomic Analysis Uncovers Genes Responsible for the DHA Enhancement in the Mutant Aurantiochytrium sp.

Comparative Transcriptomic Analysis Uncovers Genes Responsible for the DHA Enhancement in the Mutant Aurantiochytrium sp.

Control Mechanism for Carbon‐Chain Length in Polyunsaturated Fatty‐Acid Synthases

Control Mechanism for cis Double‐Bond Formation by Polyunsaturated Fatty‐Acid Synthases

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