scholarly journals Engineering isoprenoid quinone production in yeast

2020 ◽  
Author(s):  
Divjot Kaur ◽  
Christophe Corre ◽  
Fabrizio Alberti
Keyword(s):  
Yeast Strain ◽  
Mevalonate Pathway ◽  
Coenzyme Q ◽  
Shikimate Pathway ◽  
Bioactive Molecules ◽  
Primary Metabolism ◽  
Isoprenoid Quinone ◽  
New Host ◽  
Domains Of Life ◽  
Isoprenoid Quinones

Isoprenoid quinones are bioactive molecules that include an isoprenoid chain and a quinone head. They are traditionally found to be involved in primary metabolism, where they act as electron transporters, but specialized isoprenoid quinones are also produced by all domains of life. Here, we report the engineering of a baker's yeast strain, Saccharomyces cerevisiae EPYFA3, for the production of isoprenoid quinones. Our yeast strain was developed through overexpression of the shikimate pathway in a well-established recipient strain (S. cerevisiae EPY300) where the mevalonate pathway is overexpressed. As a proof of concept, our new host strain was used to overproduce the endogenous isoprenoid quinone coenzyme Q6, resulting in a final four-fold production increase. EPYFA3 represents a valuable platform for the heterologous production of high value isoprenoid quinones. EPYFA3 will also facilitate the elucidation of isoprenoid quinone biosynthetic pathways.

scholarly journals The Mevalonate Pathway of Staphylococcus aureus

2008 ◽  
Vol 191 (3) ◽  
pp. 851-861 ◽  
Author(s):  
Carl J. Balibar ◽  
Xiaoyu Shen ◽  
Jianshi Tao
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Dna Microarrays ◽  
Mevalonate Pathway ◽  
Inducible Promoter ◽  
Primary Metabolism ◽  
Gram Positive Bacteria ◽  
Cell Surface Structures ◽  
Transcriptional Changes ◽  
Electron Carriers

ABSTRACT Isoprenoids are a class of ubiquitous organic molecules synthesized from the five-carbon starter unit isopentenyl pyrophosphate (IPP). Comprising more than 30,000 known natural products, isoprenoids serve various important biological functions in many organisms. In bacteria, undecaprenyl pyrophosphate is absolutely required for the formation of cell wall peptidoglycan and other cell surface structures, while ubiquinones and menaquinones, both containing an essential prenyl moiety, are key electron carriers in respiratory energy generation. There is scant knowledge on the nature and regulation of bacterial isoprenoid pathways. In order to explore the cellular responses to perturbations in the mevalonate pathway, responsible for producing the isoprenoid precursor IPP in many gram-positive bacteria and eukaryotes, we constructed three strains of Staphylococcus aureus in which each of the mevalonate pathway genes is regulated by an IPTG (isopropyl-β-d-thiogalactopyranoside)-inducible promoter. We used DNA microarrays to profile the transcriptional effects of downregulating the components of the mevalonate pathway in S. aureus and demonstrate that decreased expression of the mevalonate pathway leads to widespread downregulation of primary metabolism genes, an upregulation in virulence factors and cell wall biosynthetic determinants, and surprisingly little compensatory expression in other isoprenoid biosynthetic genes. We subsequently correlate these transcriptional changes with downstream metabolic consequences.

scholarly journals Coenzyme Q10 Biosynthesis Established in the Non-Ubiquinone Containing Corynebacterium glutamicum by Metabolic Engineering

2021 ◽  
Vol 9 ◽  
Author(s):  
Arthur Burgardt ◽  
Ayham Moustafa ◽  
Marcus Persicke ◽  
Jens Sproß ◽  
Thomas Patschkowski ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Corynebacterium Glutamicum ◽  
Paracoccus Denitrificans ◽  
Coenzyme Q ◽  
Shikimate Pathway ◽  
Farnesyl Diphosphate ◽  
Mass Spectrometry Analysis ◽  
Biotechnological Production ◽  
Spectrometry Analysis ◽  
Shake Flask Cultivation

Coenzyme Q10 (CoQ10) serves as an electron carrier in aerobic respiration and has become an interesting target for biotechnological production due to its antioxidative effect and benefits in supplementation to patients with various diseases. For the microbial production, so far only bacteria have been used that naturally synthesize CoQ10 or a related CoQ species. Since the whole pathway involves many enzymatic steps and has not been fully elucidated yet, the set of genes required for transfer of CoQ10 synthesis to a bacterium not naturally synthesizing CoQ species remained unknown. Here, we established CoQ10 biosynthesis in the non-ubiquinone-containing Gram-positive Corynebacterium glutamicum by metabolic engineering. CoQ10 biosynthesis involves prenylation and, thus, requires farnesyl diphosphate as precursor. A carotenoid-deficient strain was engineered to synthesize an increased supply of the precursor molecule farnesyl diphosphate. Increased farnesyl diphosphate supply was demonstrated indirectly by increased conversion to amorpha-4,11-diene. To provide the first CoQ10 precursor decaprenyl diphosphate (DPP) from farnesyl diphosphate, DPP synthase gene ddsA from Paracoccus denitrificans was expressed. Improved supply of the second CoQ10 precursor, para-hydroxybenzoate (pHBA), resulted from metabolic engineering of the shikimate pathway. Prenylation of pHBA with DPP and subsequent decarboxylation, hydroxylation, and methylation reactions to yield CoQ10 was achieved by expression of ubi genes from Escherichia coli. CoQ10 biosynthesis was demonstrated in shake-flask cultivation and verified by liquid chromatography mass spectrometry analysis. To the best of our knowledge, this is the first report of CoQ10 production in a non-ubiquinone-containing bacterium.

scholarly journals Glucose-1,6-bisphosphate, a key metabolic regulator, is synthesized by a distinct family of α-phosphohexomutases widely distributed in prokaryotes

2021 ◽  
Author(s):  
Niels Neumann ◽  
Simon Friz ◽  
Karl Forchhammer
Keyword(s):  
Phylogenetic Analysis ◽  
Essential Role ◽  
Biochemical Analysis ◽  
Reverse Reaction ◽  
Primary Metabolism ◽  
Human Gut ◽  
Unicellular Cyanobacterium ◽  
Domains Of Life ◽  
Fructose 1,6 Bisphosphate ◽  
Pcc 6803

AbstractThe reactions of α-D-phosphohexomutases (αPHM) are ubiquitous, key to primary metabolism and essential for several processes in all domains of life. The functionality of these enzymes relies on an initial auto-phosphorylation step which requires the presence of α-D-glucose-1,6-bisphosphate (Glc-1,6-BP). While well investigated in vertebrates, the origin of this activator compound in bacteria is unknown. Here we show that the Slr1334 protein from the unicellular cyanobacterium Synechocysitis sp. PCC 6803 is a Glc-1,6-BP-synthase. Biochemical analysis revealed that Slr1334 efficiently converts fructose-1,6-bisphosphate and α-D-glucose-1-phosphate/α-D-glucose-6-phosphate into Glc-1,6-BP and also catalyzes the reverse reaction. Phylogenetic analysis revealed that the slr1334 product belongs to a primordial subfamily of αPHMs that is present especially in deeply branching bacteria and also includes human commensals and pathogens. Interestingly, the homologue of Slr1334 in the human gut bacterium Bacteroides salyersiae catalyzes the same reaction, suggesting a conserved and essential role for the members of this αPHM subfamily.

scholarly journals The pattern and control of isoprenoid quinone and tocopherol metabolism in the germinating grain of wheat (Triticum vulgare)

1968 ◽  
Vol 108 (3) ◽  
pp. 475-482 ◽  
Author(s):  
G. S. Hall ◽  
D L Laidman
Keyword(s):  
Gibberellic Acid ◽  
Biological Significance ◽  
Isoprenoid Quinone ◽  
Wheat Grain ◽  
Triticum Vulgare ◽  
Aleurone Cells ◽  
Wheat Shoot ◽  
Isoprenoid Quinones ◽  
And Control

1. The syntheses of ubiquinone-9 and plastoquinone-9 were used as parameters respectively of mitochondrial and proplastid development in the germinating wheat grain. 2. The changes in the amounts of the tocopherols were also studied and the possible biological significance of these changes is discussed. During germination, the dimethyl tocopherols of the resting grain are probably not utilized for the synthesis of α-tocopherol. 3. It was demonstrated that ubiquinone synthesis, and hence probably mitochondrial development, in the aleurone cells during germination, is independent of control by gibberellic acid from the embryo. 4. The influence of light on the syntheses of the isoprenoid quinones in the etiolated wheat shoot was investigated. In particular, illumination did not stimulate the synthesis of either α-tocopherol or α-tocopherolquinone.

scholarly journals BIOMASS YIELD 1 regulates sorghum biomass and grain yield via the shikimate pathway

2020 ◽  
Vol 71 (18) ◽  
pp. 5506-5520 ◽  
Author(s):  
Jun Chen ◽  
Mengjiao Zhu ◽  
Ruixiang Liu ◽  
Meijing Zhang ◽  
Ya Lv ◽  
...  
Keyword(s):  
Grain Yield ◽  
Molecular Mechanisms ◽  
Histological Analysis ◽  
Biomass Yield ◽  
Agronomic Traits ◽  
Shikimate Pathway ◽  
Floral Organ ◽  
Primary Metabolism ◽  
Primary And Secondary Metabolism ◽  
The Relationship

Abstract Biomass and grain yield are key agronomic traits in sorghum (Sorghum bicolor); however, the molecular mechanisms that regulate these traits are not well understood. Here, we characterized the biomass yield 1 (by1) mutant, which displays a dramatically altered phenotype that includes reduced plant height, narrow stems, erect and narrow leaves, and abnormal floral organs. Histological analysis suggested that these phenotypic defects are mainly caused by inhibited cell elongation and abnormal floral organ development. Map-based cloning revealed that BY1 encodes a 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS) that catalyses the first step of the shikimate pathway. BY1 was localized in chloroplasts and was ubiquitously distributed in the organs examined, particularly in the roots, stems, leaves, and panicles, which was consistent with its role in biomass production and grain yield. Transcriptome analysis and metabolic profiling revealed that BY1 was involved in primary metabolism and that it affected the biosynthesis of various secondary metabolites, especially flavonoids. Taken together, these findings demonstrate that BY1 affects biomass and grain yield in sorghum by regulating primary and secondary metabolism via the shikimate pathway. Moreover, our results provide important insights into the relationship between plant development and metabolism.

scholarly journals Conversion of Mevalonate 3-Kinase into 5-Phosphomevalonate 3-Kinase by Single Amino Acid Mutations

2019 ◽  
Vol 85 (9) ◽  
Author(s):  
Kento Motoyama ◽  
Fumiaki Sobue ◽  
Hiroshi Kawaide ◽  
Tohru Yoshimura ◽  
Hisashi Hemmi
Keyword(s):  
Amino Acid ◽  
Mevalonate Pathway ◽  
Bioactive Molecules ◽  
Single Amino Acid ◽  
Carotenoid Pigment ◽  
Bacterial Cells ◽  
Content Type ◽  
Single Amino Acid Residue ◽  
Amino Acid Mutations ◽  
Thermophilic Archaea

ABSTRACT Mevalonate 3-kinase plays a key role in a recently discovered modified mevalonate pathway specific to thermophilic archaea of the order Thermoplasmatales. The enzyme is homologous to diphosphomevalonate decarboxylase, which is involved in the widely distributed classical mevalonate pathway, and to phosphomevalonate decarboxylase, which is possessed by halophilic archaea and some Chloroflexi bacteria. Mevalonate 3-kinase catalyzes the ATP-dependent 3-phosphorylation of mevalonate but does not catalyze the subsequent decarboxylation as related decarboxylases do. In this study, a substrate-interacting glutamate residue of Thermoplasma acidophilum mevalonate 3-kinase was replaced by smaller amino acids, including its counterparts in diphosphomevalonate decarboxylase and phosphomevalonate decarboxylase, with the aim of altering substrate specificity. These single amino acid mutations resulted in the conversion of mevalonate 3-kinase into 5-phosphomevalonate 3-kinase, which can synthesize 3,5-bisphosphomevalonate from 5-phosphomevalonate. The mutants catalyzing the hitherto undiscovered reaction enabled the construction of an artificial mevalonate pathway in Escherichia coli cells, as was demonstrated by the accumulation of lycopene, a red carotenoid pigment. IMPORTANCE Isoprenoid is the largest family of natural compounds, including important bioactive molecules such as vitamins, hormones, and natural medicines. The mevalonate pathway is a target for metabolic engineering because it supplies precursors for isoprenoid biosynthesis. Mevalonate 3-kinase is an enzyme involved in the modified mevalonate pathway specific to limited species of thermophilic archaea. Replacement of a single amino acid residue in the active site of the enzyme changed its substrate preference and allowed the mutant enzymes to catalyze a previously undiscovered reaction. Using the genes encoding the mutant enzymes and other archaeal enzymes, we constructed an artificial mevalonate pathway, which can produce the precursor of isoprenoid through an unexplored route, in bacterial cells.

Kaempferol as a precursor for ubiquinone (coenzyme Q) biosynthesis: An atypical node between specialized metabolism and primary metabolism

2022 ◽  
Vol 66 ◽  
pp. 102165
Author(s):  
Antoine Berger ◽  
Scott Latimer ◽  
Lauren R. Stutts ◽  
Eric Soubeyrand ◽  
Anna K. Block ◽  
...  
Keyword(s):  
Coenzyme Q ◽  
Primary Metabolism ◽  
Specialized Metabolism

scholarly journals Unusual features of the c-ring of F1FO ATP synthases

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
A. V. Vlasov ◽  
K. V. Kovalev ◽  
S.-H. Marx ◽  
E. S. Round ◽  
I. Yu. Gushchin ◽  
...  
Keyword(s):  
High Resolution ◽  
Molecular Mechanisms ◽  
Coenzyme Q ◽  
Ion Leakage ◽  
Vis Spectroscopy ◽  
High Resolution Structure ◽  
Isoprenoid Quinones ◽  
Electron Carriers ◽  
Resolution Structure ◽  
Atp Synthases

AbstractMembrane integral ATP synthases produce adenosine triphosphate, the universal “energy currency” of most organisms. However, important details of proton driven energy conversion are still unknown. We present the first high-resolution structure (2.3 Å) of the in meso crystallized c-ring of 14 subunits from spinach chloroplasts. The structure reveals molecular mechanisms of intersubunit contacts in the c14-ring, and it shows additional electron densities inside the c-ring which form circles parallel to the membrane plane. Similar densities were found in all known high-resolution structures of c-rings of F1FO ATP synthases from archaea and bacteria to eukaryotes. The densities might originate from isoprenoid quinones (such as coenzyme Q in mitochondria and plastoquinone in chloroplasts) that is consistent with differential UV-Vis spectroscopy of the c-ring samples, unusually large distance between polar/apolar interfaces inside the c-ring and universality among different species. Although additional experiments are required to verify this hypothesis, coenzyme Q and its analogues known as electron carriers of bioenergetic chains may be universal cofactors of ATP synthases, stabilizing c-ring and prevent ion leakage through it.

scholarly journals Isopentenyl Phosphate Kinases are Ubiquitous and Copy Numbers are Conserved in Plant Genomes

2019 ◽  
Author(s):  
Ben Hu ◽  
Heng Yao ◽  
Feng Li ◽  
Ran Wang ◽  
Yalong Xu ◽  
...  
Keyword(s):  
Plant Species ◽  
Mevalonate Pathway ◽  
Isoprenoid Biosynthesis ◽  
Plant Genome ◽  
Peptide Sequence ◽  
Plant Genomes ◽  
Copy Numbers ◽  
Key Enzyme ◽  
Domains Of Life ◽  
Isopentenyl Phosphate

Abstract Key message: Isopentenyl phosphate kinase (IPK) is a key enzyme in mevalonate pathway in isoprenoid biosynthesis. We analyzed 37 presumptive IPK sequences from 35 plants. An specific evolution model was found in plant IPKs, which can be used as an new target in studying the plant isoprenoids metabilte. Abstract: Isopentenyl phosphate kinase (IPK) is a recently discovered enzyme played key role in mevalonate pathway in isoprenoid biosynthesis. Here, we showed that IPKs are ubiquitously present in plant genomes. All IPKs previously identified had AAK domain. From 35 plant species with genome assembly data available, we extracted all AAK family members. Using OrthoMCL, we identified a group of 37 sequences in which Arabidopsis IPK was included. Further analysis showed that each peptide sequence in this group has a His residue which is a signature of IPK enzyme, indicating that the genes in this group were IPKs. Not like these in other domains of life which showed spotty distribution over the tree of life, virtually all plant genomes we analyzed here had IPK genes. Further, copy numbers of IPKs were very conserved in that no higher than 2 copies remained in each plant genome. Plant IPKs formed a distinctive clade in phylogenetic tree of plant AAK gene family, and had a phylogenetic topology conformed to that of plant species. The IPKs we identified here would provide new molecular targets for characterization of plant mevalonate pathway, and shed light on biochemistry of plant isoprenoids biosynthesis.

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