Expanding the product portfolio of fungal type I fatty acid synthases

Zhu, Zhiwei; Zhou, Yongjin J.; Krivoruchko, Anastasia; Grininger, Martin; Zhao, Zongbao K.; Nielsen, Jens

doi:10.1038/nchembio.2301

Cited by 105 publications

(85 citation statements)

References 25 publications

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“…Structural knowledge on the interaction landscape of ACP within type I FAS and understanding the mechanism of ACP-mediated substrate shuttling are highly sought after in antibiotic development [19][20][21] and biofuel production efforts. [22][23][24] This study demonstrates that ACP interaction with a catalytic center can be modulated by stalling catalysis at another reaction site, suggesting that the loading state of ACP's phosphopantetheine arm may have at least a partial role in determination of ACP localization within the reaction chambers of type I fungal FAS. Future structural studies should systematically investigate ACP distribution upon inhibition of each reaction site in FAS (Supplementary Figure 1A) using site-specific inhibitors or point mutations in recombinantly expressed enzymes.…”

mentioning

confidence: 77%

Electron cryomicroscopy observation of acyl carrier protein translocation in type I fungal fatty acid synthase

Lou

Iyer

Hasan

et al. 2019

Preprint

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During fatty acid biosynthesis, acyl carrier proteins (ACPs) from type I fungal fatty acid synthase (FAS) shuttle substrates and intermediates within a reaction chamber that hosts multiple spatially-fixed catalytic centers. A major challenge in understanding the mechanism of ACP-mediated substrate shuttling is experimental observation of its transient interaction landscape within the reaction chamber. Here, we have shown that ACP spatial distribution is sensitive to the presence of substrates in a catalytically inhibited state, which enables high-resolution investigation of the ACP-dependent conformational transitions within the enoyl reductase (ER) reaction site. In two fungal FASs with distinct ACP localization, the shuttling domain is targeted to the ketoacyl-synthase (KS) domain and away from other catalytic centers, such as acetyl-transferase (AT) and ER domains by steric blockage

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mentioning

confidence: 77%

Electron cryomicroscopy observation of acyl carrier protein translocation in type I fungal fatty acid synthase

Lou

Iyer

Hasan

et al. 2019

Preprint

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“…Potentially, the introduction of a constitutively neutral charge or a smaller AA at site K173 generates more space for the growing acyl chain within the FAS-chamber, thus increasing the amount of C18 FAs at the expense of C16 FAs. However, a recent study presented that it was possible to add a complete thioesterase domain at the Cterminal end of the S. cerevisiae FAS next to the ACP domain without causing loss of function (Zhu et al, 2017). In order to determine if the observed difference in FA chain-length is the result of a regulatory acetylation on K173 or a steric alteration of the catalytic chamber when introducing a non-charged/smaller AA, further studies are required.…”

Section: Discussionmentioning

confidence: 99%

Investigation of putative regulatory acetylation sites in Fas2p ofSaccharomyces cerevisiae

Bergman

Wenning

Siewers

et al. 2018

Preprint

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Yeast metabolism is highly regulated, in part via coordinated reprogramming of metabolism on a transcriptional level, for example in response to environmental changes. Furthermore, regulation occurs on the protein level via posttranslational modifications directly affecting enzymatic activity -a mode of regulation that has the benefit of being very fast in response to environmental changes. One group of posttranslational modification that has been suggested to have a high impact on regulation of metabolism are acetylations. Around 4000 distinct protein acetylation sites have been found in Saccharomyces cerevisiae, many of which are located in central metabolic enzymes. However, reports on the verification of regulatory roles of specific acetylation sites on these metabolic enzymes have yet to emerge. This study investigates putative regulatory acetylation sites on Fas2p, which in concert with Fas1p is responsible for cytosolic fatty acid (FA) biosynthesis in S. cerevisiae. Fas2p stands out as one of the most highly acetylated proteins in yeast and is located at a branchpoint of acetyl-CoA metabolism. The amino acids (AAs) glutamine (Q) and arginine (R) were introduced to mimic a constitutively acetylated or non-acetylatable state at three separate lysine sites (K) (K83, K173 and K1551) confirmed to be acetylated in two independent studies, either separately or simultaneously. The results suggest that the residue replacement system in the specific case interferes with the enzymatic function of the fatty acid synthase (FAS), as QQQ and RRR triple mutants both reduce the amount of secreted free fatty acids (FFAs) in a faa1∆ faa4∆ yeast deletion mutant. The K173Q substitution significantly decreased C16 FA species at the expense of C18 FAs, while no such change could be observed for the corresponding K173R modification.3 KeywordsSaccharomyces cerevisiae, fatty acid synthase, regulation, protein acetylation, residue replacement system cytosolic/peroxisomal glyoxylate cycle. This pathway is essential for growth on C2 compounds (like acetate and ethanol), but also for degradation of FAs via β-oxidation, and produces succinate which can enter the TCA cycle .As a central metabolite, acetyl-CoA is an important building block in many anabolic reactions, but it also plays a role in the posttranslational modification of proteins by serving as a substrate for lysine acetyltransferases (KATs), which catalyze the transfer of acetyl groups to the epsilon-amino groups of lysines (Lys, K). In this way, fluctuating concentrations of acetyl-CoA, which are connected to the metabolic condition of the cell, are translated into dynamic protein acetylations that regulate a range of cellular functions. Enzymes that remove acetylations from proteins are called lysine deacetylases (KDACs). So far, seven KATs and nine KDACs, respectively, have been identified in yeast. KATs are classified based on their cellular localization, with being either of the nuclear type (A-type) or the cytoplasmic type (Btype), whereas KDACs are phylogenetically div...

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“…By adapting active site specificities, mutations essentially steer de novo fatty acid synthesis towards the early release of not yet fully elongated C 16 and C 18 -acyl-CoA, while leaving the overall molecular mechanisms intact. Indeed, evaluated on the basis of SCFA yields, the approach turned out to be highly powerful compared to other strategies that were overwriting native synthesis with a short-chain acyl-ACP specific thioesterase that is inserted as extra domain into the polypeptide chain [60–63]. …”

Section: Reviewmentioning

confidence: 99%

Strategies in megasynthase engineering – fatty acid synthases (FAS) as model proteins

Fischer¹,

Grininger²

2017

Beilstein J. Org. Chem.

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Megasynthases are large multienzyme proteins that produce a plethora of important natural compounds by catalyzing the successive condensation and modification of precursor units. Within the class of megasynthases, polyketide synthases (PKS) are responsible for the production of a large spectrum of bioactive polyketides (PK), which have frequently found their way into therapeutic applications. Rational engineering approaches have been performed during the last 25 years that seek to employ the “assembly-line synthetic concept” of megasynthases in order to deliver new bioactive compounds. Here, we highlight PKS engineering strategies in the light of the newly emerging structural information on megasynthases, and argue that fatty acid synthases (FAS) are and will be valuable objects for further developing this field.

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Expanding the product portfolio of fungal type I fatty acid synthases

Cited by 105 publications

References 25 publications

Electron cryomicroscopy observation of acyl carrier protein translocation in type I fungal fatty acid synthase

Electron cryomicroscopy observation of acyl carrier protein translocation in type I fungal fatty acid synthase

Investigation of putative regulatory acetylation sites in Fas2p ofSaccharomyces cerevisiae

Strategies in megasynthase engineering – fatty acid synthases (FAS) as model proteins

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