scholarly journals Transcriptome-wide Identification and Quantification of Caffeoylquinic Acid Biosynthesis Pathway and Prediction of their Putative BAHDs Gene Complex in A. spathulifolius

2021 ◽  
Author(s):  
SeonJoo Park ◽  
Jean Claude Sivagami ◽  
Sunmi Park
Keyword(s):  
Phenylpropanoid Pathway ◽  
Biosynthesis Pathway ◽  
Gene Complex ◽  
Biotic And Abiotic Stress ◽  
Acid Biosynthesis ◽  
Gene Encoding ◽  
Caffeoylquinic Acid ◽  
Key Enzyme ◽  
By Products

The phenylpropanoid pathway is a major secondary metabolite pathway that helps plants overcome biotic and abiotic stress and produces various by-products that promote human health. Its byproduct, chloroquinic acid (CQA), is a soluble phenolic compound present in many angiosperms. Hy-droxycinnamate-CoA shikimate/quinate transferase(BAHDs superfamily enzyme) is a significant en-zyme that plays a role in accumulating CQA biosynthesis. This study analyzed transcriptome-wide identification of the phenylpropanoid to chloroquinic acid biosynthesis candidate genes in A. spathulifolius flowers and leaves. Transcriptomic analyses of the flowers and leaves showed a differential expression of the PPP and CQA biosynthesis regulated unigenes. An analysis of PPP captive unigenes revealed the following: the major duplication of the key enzyme, PAL, 120 unigenes in leaves and 76 in flowers; the gene encoding C3’H, 169 unigenes in leaves and 140 unigenes in flowers; duplicated unigenes of 4CL, 41 in leaves and 27 in flowers. In addition, C4H unigenes had 12 unigenes in the leaves of A. spathulifolius and four in the flowers. The characterization of the BAHDs superfamily members identified 82 in leaves and 72 in flowers. Among them, phylogenetic analysis showed that five unigenes encoded HQT and three en-coded HCT in A. spathulifolius. The three HQT are common to both leaves and flowers, whereas the two HQT were specialized for leaves. The pattern of HQT synthesis was upregulated in flowers, whereas HCT was expressed strongly in the leaves of A. spathulifolius. Overall, 4CL, C4H, and HQT are expressed strongly in flowers, and caffeic acid and HCT show more expression in leaves. Therefore, CQA biosynthesis occurs in the flowers of A. spathulifolius rather than leaves.

scholarly journals Transcriptome-Wide Identification and Quantification of Caffeoylquinic Acid Biosynthesis Pathway and Prediction of Its Putative BAHDs Gene Complex in A. spathulifolius

2021 ◽  
Vol 22 (12) ◽  
pp. 6333
Author(s):  
Sivagami-Jean Claude ◽  
Sunmi Park ◽  
Seon-Joo Park
Keyword(s):  
Phylogenetic Analysis ◽  
Abiotic Stress ◽  
Human Health ◽  
Caffeic Acid ◽  
Phenylpropanoid Pathway ◽  
Biosynthesis Pathway ◽  
Gene Complex ◽  
Biotic And Abiotic Stress ◽  
Acid Biosynthesis ◽  
Caffeoylquinic Acid

The phenylpropanoid pathway is a major secondary metabolite pathway that helps plants overcome biotic and abiotic stress and produces various byproducts that promote human health. Its byproduct caffeoylquinic acid is a soluble phenolic compound present in many angiosperms. Hydroxycinnamate-CoA shikimate/quinate transferase is a significant enzyme that plays a role in accumulating CQA biosynthesis. This study analyzed transcriptome-wide identification of the phenylpropanoid to caffeoylquinic acid biosynthesis candidate genes in A. spathulifolius flowers and leaves. Transcriptomic analyses of the flowers and leaves showed a differential expression of the PPP and CQA biosynthesis regulated unigenes. An analysis of PPP-captive unigenes revealed a major duplication in the following genes: PAL, 120 unigenes in leaves and 76 in flowers; C3′H, 169 unigenes in leaves and 140 in flowers; 4CL, 41 unigenes in leaves and 27 in flowers; and C4H, 12 unigenes in leaves and 4 in flowers. The phylogenetic analysis revealed 82 BAHDs superfamily members in leaves and 72 in flowers, among which five unigenes encode for HQT and three for HCT. The three HQT are common to both leaves and flowers, whereas the two HQT were specialized for leaves. The pattern of HQT synthesis was upregulated in flowers, whereas HCT was expressed strongly in the leaves of A. spathulifolius. Overall, 4CL, C4H, and HQT are expressed strongly in flowers and CAA and HCT show more expression in leaves. As a result, the quantification of HQT and HCT indicates that CQA biosynthesis is more abundant in the flowers and synthesis of caffeic acid in the leaves of A. spathulifolius.

scholarly journals Characterization of Ligand Binding to the Bifunctional Key Enzyme in the Sialic Acid Biosynthesis by NMR

2004 ◽  
Vol 279 (53) ◽  
pp. 55722-55727 ◽  
Author(s):  
Andrew J. Benie ◽  
Astrid Blume ◽  
Richard R. Schmidt ◽  
Werner Reutter ◽  
Stephan Hinderlich ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Ligand Binding ◽  
Acid Biosynthesis ◽  
Key Enzyme

scholarly journals Isolation and Characterization of FecA- and FeoB-Mediated Iron Acquisition Systems of the Spirochete Leptospira biflexa by Random Insertional Mutagenesis

2005 ◽  
Vol 187 (9) ◽  
pp. 3249-3254 ◽  
Author(s):  
Hélène Louvel ◽  
Isabelle Saint Girons ◽  
Mathieu Picardeau
Keyword(s):  
Insertional Mutagenesis ◽  
Iron Acquisition ◽  
Transposon Mutagenesis ◽  
Ecological Niches ◽  
Acid Biosynthesis ◽  
Gene Encoding ◽  
Isolation And Characterization ◽  
Transposon Mutants

ABSTRACT The specific mechanisms by which Leptospira spp. acquire iron from their ecological niches are unknown. A major factor contributing to our ignorance of spirochetal biology is the lack of methods for genetic analysis of these organisms. In this study, we have developed a system for random transposon mutagenesis of Leptospira biflexa using a mariner transposon, Himar1. To demonstrate the validity of Himar1 in vivo transposon mutagenesis in L. biflexa, a screen of mutants for clones impaired in amino acid biosynthesis was first performed, enabling the identification of tryptophan and glutamate auxotrophs. To investigate iron transporters, 2,000 L. biflexa transposon mutants were screened onto media with and without hemin, thus allowing the identification of five hemin-requiring mutants, and the putative genes responsible for this phenotype were identified. Three mutants had distinct insertions in a gene encoding a protein which shares homology with the TonB-dependent receptor FecA, involved in ferric citrate transport. We also identified two mutants with a Himar1 insertion into a feoB-like gene, the product of which is required for ferrous iron uptake in many bacterial organisms. Interestingly, the growth inhibition exhibited by the fecA and feoB mutants was relieved by deferoxamine, suggesting the presence of a ferric hydroxamate transporter. These results confirm the importance of iron for the growth of Leptospira and its ability to use multiple iron sources.

scholarly journals Purification, Overproduction, and Partial Characterization of β-RFAP Synthase, a Key Enzyme in the Methanopterin Biosynthesis Pathway†

2002 ◽  
Vol 184 (16) ◽  
pp. 4442-4448 ◽  
Author(s):  
Joseph W. Scott ◽  
Madeline E. Rasche
Keyword(s):  
Biosynthetic Pathway ◽  
Methanogenic Archaea ◽  
Methylotrophic Bacteria ◽  
Methylobacterium Extorquens ◽  
Gene Encoding ◽  
Methanosarcina Thermophila ◽  
Inorganic Pyrophosphate ◽  
Sulfate Reducing ◽  
Key Enzyme

ABSTRACT Methanopterin is a folate analog involved in the C1 metabolism of methanogenic archaea, sulfate-reducing archaea, and methylotrophic bacteria. Although a pathway for methanopterin biosynthesis has been described in methanogens, little is known about the enzymes and genes involved in the biosynthetic pathway. The enzyme β-ribofuranosylaminobenzene 5′-phosphate synthase (β-RFAP synthase) catalyzes the first unique step to be identified in the pathway of methanopterin biosynthesis, namely, the condensation of p-aminobenzoic acid with phosphoribosylpyrophosphate to form β-RFAP, CO2, and inorganic pyrophosphate. The enzyme catalyzing this reaction has not been purified to homogeneity, and the gene encoding β-RFAP synthase has not yet been identified. In the present work, we report on the purification to homogeneity of β-RFAP synthase. The enzyme was purified from the methane-producing archaeon Methanosarcina thermophila, and the N-terminal sequence of the protein was used to identify corresponding genes from several archaea, including the methanogen Methanococcus jannaschii and the sulfate-reducing archaeon Archaeoglobus fulgidus. The putative β-RFAP synthase gene from A. fulgidus was expressed in Escherichia coli, and the enzymatic activity of the recombinant gene product was verified. A BLAST search using the deduced amino acid sequence of the β-RFAP synthase gene identified homologs in additional archaea and in a gene cluster required for C1 metabolism by the bacterium Methylobacterium extorquens. The identification of a gene encoding a potential β-RFAP synthase in M. extorquens is the first report of a putative methanopterin biosynthetic gene found in the Bacteria and provides evidence that the pathways of methanopterin biosynthesis in Bacteria and Archaea are similar.

scholarly journals Characterization of Ligand Binding to the Bifunctional Key Enzyme in the Sialic Acid Biosynthesis by NMR

2004 ◽  
Vol 279 (53) ◽  
pp. 55715-55721 ◽  
Author(s):  
Astrid Blume ◽  
Andrew J. Benie ◽  
Florian Stolz ◽  
Richard R. Schmidt ◽  
Werner Reutter ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Ligand Binding ◽  
Acid Biosynthesis ◽  
Key Enzyme

scholarly journals Inactivation of the ampDE Operon Increases Transcription of algD and Affects Morphology and Encystment of Azotobacter vinelandii

2000 ◽  
Vol 182 (17) ◽  
pp. 4829-4835 ◽  
Author(s):  
Cinthia Núñez ◽  
Soledad Moreno ◽  
Luis Cárdenas ◽  
Gloria Soberón-Chávez ◽  
Guadalupe Espín
Keyword(s):  
Escherichia Coli ◽  
Biosynthetic Pathway ◽  
Azotobacter Vinelandii ◽  
Transmembrane Protein ◽  
Nucleotide Sequencing ◽  
Insertion Mutant ◽  
Gene Encoding ◽  
Alginate Production ◽  
Key Enzyme

ABSTRACT Transcription of algD, encoding GDP-mannose dehydrogenase, the key enzyme in the alginate biosynthetic pathway, is highly regulated in Azotobacter vinelandii. We describe here the characterization of a Tn5 insertion mutant (AC28) which shows a higher level of expression of analgD::lacZ fusion. AC28 cells were morphologically abnormal and unable to encyst. The cloning and nucleotide sequencing of the Tn5-disrupted locus in AC28 revealed an operon homologous to the Escherichia coli ampDEoperon. Tn5 was located within the ampD gene, encoding a cytosolicN-acetyl-anhydromuramyl-l-alanine amidase that participates in the intracellular recycling of peptidoglycan fragments. The ampE gene encodes a transmembrane protein, but the function of the protein is not known. We constructed strains carryingampD or ampE mutations and one with anampDE deletion. The strain with a deletion of theampDE operon showed a phenotype similar to that of mutant AC28. The present work demonstrates that both alginate production and bacterial encystment are greatly influenced by the bacterial ability to recycle its cell wall.

scholarly journals Characterization of the Caulobacter crescentus Holdfast Polysaccharide Biosynthesis Pathway Reveals Significant Redundancy in the Initiating Glycosyltransferase and Polymerase Steps

2008 ◽  
Vol 190 (21) ◽  
pp. 7219-7231 ◽  
Author(s):  
Evelyn Toh ◽  
Harry D. Kurtz ◽  
Yves V. Brun
Keyword(s):  
Caulobacter Crescentus ◽  
Repeat Unit ◽  
Deletion Mutants ◽  
Biosynthesis Pathway ◽  
Surface Adhesion ◽  
Wild Type ◽  
Chitin Deacetylase ◽  
Gene Encoding ◽  
Polysaccharide Biosynthesis

ABSTRACT Caulobacter crescentus cells adhere to surfaces by using an extremely strong polar adhesin called the holdfast. The polysaccharide component of the holdfast is comprised in part of oligomers of N-acetylglucosamine. The genes involved in the export of the holdfast polysaccharide and the anchoring of the holdfast to the cell were previously discovered. In this study, we identified a cluster of polysaccharide biosynthesis genes (hfsEFGH) directly adjacent to the holdfast polysaccharide export genes. Sequence analysis indicated that these genes are involved in the biosynthesis of the minimum repeat unit of the holdfast polysaccharide. HfsE is predicted to be a UDP-sugar lipid-carrier transferase, the glycosyltransferase that catalyzes the first step in polysaccharide biosynthesis. HfsF is predicted to be a flippase, HfsG is a glycosyltransferase, and HfsH is similar to a polysaccharide (chitin) deacetylase. In-frame hfsG and hfsH deletion mutants resulted in severe deficiencies both in surface adhesion and in binding to the holdfast-specific lectin wheat germ agglutinin. In contrast, hfsE and hfsF mutants exhibited nearly wild-type levels of adhesion and holdfast synthesis. We identified three paralogs to hfsE, two of which are redundant to hfsE for holdfast synthesis. We also identified a redundant paralog to the hfsC gene, encoding the putative polysaccharide polymerase, and present evidence that the hfsE and hfsC paralogs, together with the hfs genes, are absolutely required for proper holdfast synthesis.

Inactivation of mshB, a key gene in the mycothiol biosynthesis pathway in Mycobacterium smegmatis

Microbiology ◽  
2003 ◽  
Vol 149 (5) ◽  
pp. 1341-1349 ◽  
Author(s):  
Mamta Rawat ◽  
Svetozar Kovacevic ◽  
Helen Billman-Jacobe ◽  
Yossef Av-Gay
Keyword(s):  
Bacterial Growth ◽  
Mycobacterium Smegmatis ◽  
Parent Strain ◽  
Stationary Phases ◽  
Growth Cycle ◽  
Biosynthesis Pathway ◽  
Phenotypic Analysis ◽  
Gene Encoding ◽  
Key Enzyme ◽  
Amide Hydrolase

The mshB gene encoding N-acetyl-1-d-myo-inosityl-2-amino-2-deoxy-α-d-glucopyranoside deacetylase (MshB) is a key enzyme in mycothiol biosynthesis. Disruption of mshB in Mycobacterium smegmatis resulted in decreased production of mycothiol (5–10 % of the parent strain mc2155) but did not abolish mycothiol synthesis completely. Complementation of the MshB− mutants with the mshB gene resulted in increased mycothiol production towards the exponential and stationary phases of the bacterial growth cycle. These results suggest that another enzyme is capable of mycothiol biosynthesis by providing N-acetylglucosaminylinositol deacetylation activity in the absence of MshB. One of the candidate enzymes capable of carrying out such reactions is the MshB orthologue mycothiol amide hydrolase, MCA. However, epichromosomal expression of mca in the MshB− mutants did not restore mycothiol levels to the level of the parent strain. Unlike other mutants, which have little or no detectable levels of mycothiol, the MshB− mutant did not exhibit increased resistance to isoniazid. However, the MshB− mutant was resistant to ethionamide. Phenotypic analysis of other mutants lacking mycothiol revealed that MshA− mutants also exhibit ethionamide resistance but that a MshC−mutant was sensitive to ethionamide, suggesting that mycothiol or its early intermediates influence ethionamide activation.

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