scholarly journals Evolutionary Implications of a Peroxidase with High Affinity for Cinnamyl Alcohols from Physcomitrium patens, a Non-Vascular Plant

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1476
Author(s):  
Teresa Martínez-Cortés ◽  
Federico Pomar ◽  
Esther Novo-Uzal
Keyword(s):  
Cell Walls ◽  
Physcomitrella Patens ◽  
Vascular System ◽  
Vascular Plant ◽  
Lignin Biosynthesis ◽  
Exchange Chromatography ◽  
Limiting Factor ◽  
Kinetic Properties ◽  
Class Iii ◽  
Cationic Exchange

Physcomitrium (Physcomitrella) patens is a bryophyte highly tolerant to different stresses, allowing survival when water supply is a limiting factor. This moss lacks a true vascular system, but it has evolved a primitive water-conducting system that contains lignin-like polyphenols. By means of a three-step protocol, including ammonium sulfate precipitation, adsorption chromatography on phenyl Sepharose and cationic exchange chromatography on SP Sepharose, we were able to purify and further characterize a novel class III peroxidase, PpaPrx19, upregulated upon salt and H2O2 treatments. This peroxidase, of a strongly basic nature, shows surprising homology to angiosperm peroxidases related to lignification, despite the lack of true lignins in P. patens cell walls. Moreover, PpaPrx19 shows catalytic and kinetic properties typical of angiosperm peroxidases involved in oxidation of monolignols, being able to efficiently use hydroxycinnamyl alcohols as substrates. Our results pinpoint the presence in P. patens of peroxidases that fulfill the requirements to be involved in the last step of lignin biosynthesis, predating the appearance of true lignin.

scholarly journals Targeting hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase for lignin modification in Brachypodium distachyon

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Juan Carlos Serrani-Yarce ◽  
Luis Escamilla-Trevino ◽  
Jaime Barros ◽  
Lina Gallego-Giraldo ◽  
Yunqiao Pu ◽  
...  
Keyword(s):  
Negative Impact ◽  
Lignin Content ◽  
Brachypodium Distachyon ◽  
Lignin Biosynthesis ◽  
Reverse Direction ◽  
Wall Structure ◽  
Kinetic Properties ◽  
Loss Of Function ◽  
Down Regulation ◽  
Lignin Modification

Abstract Background Hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT) is a central enzyme of the so-called “esters” pathway to monolignols. As originally envisioned, HCT functions twice in this pathway, to form coumaroyl shikimate and then, in the “reverse” direction, to convert caffeoyl shikimate to caffeoyl CoA. The discovery of a caffeoyl shikimate esterase (CSE) that forms caffeic acid directly from caffeoyl shikimate calls into question the need for the reverse HCT reaction in lignin biosynthesis. Loss of function of HCT gives severe growth phenotypes in several dicot plants, but less so in some monocots, questioning whether this enzyme, and therefore the shikimate shunt, plays the same role in both monocots and dicots. The model grass Brachypodium distachyon has two HCT genes, but lacks a classical CSE gene. This study was therefore conducted to evaluate the utility of HCT as a target for lignin modification in a species with an “incomplete” shikimate shunt. Results The kinetic properties of recombinant B. distachyon HCTs were compared with those from Arabidopsis thaliana, Medicago truncatula, and Panicum virgatum (switchgrass) for both the forward and reverse reactions. Along with two M. truncatula HCTs, B. distachyon HCT2 had the least kinetically unfavorable reverse HCT reaction, and this enzyme is induced when HCT1 is down-regulated. Down regulation of B. distachyon HCT1, or co-down-regulation of HCT1 and HCT2, by RNA interference led to reduced lignin levels, with only modest changes in lignin composition and molecular weight. Conclusions Down-regulation of HCT1, or co-down-regulation of both HCT genes, in B. distachyon results in less extensive changes in lignin content/composition and cell wall structure than observed following HCT down-regulation in dicots, with little negative impact on biomass yield. Nevertheless, HCT down-regulation leads to significant improvements in biomass saccharification efficiency, making this gene a preferred target for biotechnological improvement of grasses for bioprocessing.

Integrated analysis of transcriptomic and metabolomic data reveals critical metabolic pathways involved in polyphenol biosynthesis in Nicotiana tabacum under chilling stress

2019 ◽  
Vol 46 (1) ◽  
pp. 30 ◽  
Author(s):  
Peilu Zhou ◽  
Qiyao Li ◽  
Guangliang Liu ◽  
Na Xu ◽  
Yinju Yang ◽  
...  
Keyword(s):  
Nicotiana Tabacum ◽  
Chilling Stress ◽  
Lignin Biosynthesis ◽  
Cinnamyl Alcohol Dehydrogenase ◽  
Integrated Analysis ◽  
Limiting Factor ◽  
Active Metabolites ◽  
Cold Temperatures ◽  
Flight Mass Spectrometry ◽  
Key Steps

Chilling stress increases the amount of polyphenols, especially lignin, which protects tobacco (Nicotiana tabacum L. cv. k326) from chilling stress. To clarify the molecular biosynthesis mechanism of the key representative compounds, specifically lignin, RNA sequencing and ultra-high pressure liquid chromatography coupled to quadrupole-time of flight mass spectrometry technologies were used to construct transcriptomic and metabolomic libraries from the leaves of tobacco plants subjected to normal (25°C) and chilling (4°C) temperature treatments. Transcriptomic libraries from the different samples were sequenced, generating more than 40million raw reads. Among nine samples, metabolomic analysis identified a total of 97 encoding enzymes that function in the key steps of pathways related to polyphenol biosynthesis, where 42 metabolites were also located. An integrated analysis of metabolic and transcriptomic data revealed that most of the intermediate metabolites and enzymes related to lignin biosynthesis were synthesised in the leaves under chilling stress, which suggests that the biosynthesis of lignin plays an important role in the response of tobacco leaves to cold temperatures. In addition, the cold insensitivity of chalcone synthase genes might be considered to be an important rate-limiting factor in the process of precursor substance flow to flavonoid biosynthesis under chilling stress. Furthermore, the upregulated expression of phenylalanine ammonia lyase (PAL), hydroxycinnamoyl transferase (HCT) and cinnamyl-alcohol dehydrogenase (CAD) under chilling stress is the key to an increase in lignin synthesis. This study provides a hypothetical basis for the screening of new active metabolites and the metabolic engineering of polyphenols in tobacco.

scholarly journals STRUCTURAL AND CHEMICAL CHARACTERISTIC OF PECTIN FROM PICEA ABIES GREENERY

2020 ◽  
pp. 59-71
Author(s):  
Evgeniy Gennad'yevich Shakhmatov ◽  
Elena Nikolayevna Makarova
Keyword(s):  
Cell Walls ◽  
New Technologies ◽  
Chemical Characteristic ◽  
Structural Features ◽  
Exchange Chromatography ◽  
Raw Material ◽  
Rhamnogalacturonan I ◽  
Wood Processing ◽  
Potential Source ◽  
Pectin Substances

The present work aimed to determine structural features of polysaccharides derived from the P. abies foliage by extraction with a (NH4)2C2O4 solution. The isolated polysaccharide was studied in detail by the methods of ion exchange chromatography, partial acidic hydrolys and NMR spectroscopy. It was shown that this polysaccharide contained polymers of various structures. The major constituents of PAO were low-methoxyl and low-acetylated 1,4-a-D-galacturonan and by minor parts of partly 2-O- and/or 3-O- acetylated rhamnogalacturonan-I (RG-I). The side carbohydrate chains of the branched region of RG-I were represented predominantly by highly branched 1,5-a-L-arabinan and minor portions of 1,4-β-D-galactan. In addition to the dominant pectins, polysaccharide PAO contained binding glycans of the glucomannans class, which indicated a close interaction of these polysaccharides in the cell walls. Thus, the structural features of pectin woody P. abies, extracted with a solution of (NH4)2C2O4, were first determined. It can be concluded that P. abies woody greens, a large tonnage waste from the wood processing industry, can be considered as a potential source of pectin substances. The results of studying the structure of components of woody green P. abies can be the basis for the development and improvement of new technologies for the integrated use of this raw material.

scholarly journals Patatin-Related Phospholipase AtpPLAIIIα Affects Lignification of Xylem in Arabidopsis and Hybrid Poplars

Plants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 451 ◽  
Author(s):  
Jin Hoon Jang ◽  
Ok Ran Lee
Keyword(s):  
Cell Walls ◽  
Lignin Content ◽  
Lignin Biosynthesis ◽  
Biofuel Production ◽  
Amide Bonds ◽  
Hybrid Poplars ◽  
Plant Lipids ◽  
Lipid Acyl Hydrolase ◽  
Specific Regulation

Lipid acyl hydrolase are a diverse group of enzymes that hydrolyze the ester or amide bonds of fatty acid in plant lipids. Patatin-related phospholipase AIIIs (pPLAIIIs) are one of major lipid acyl hydrolases that are less closely related to potato tuber patatins and are plant-specific. Recently, overexpression of ginseng-derived PgpPLAIIIβ was reported to be involved in the reduced level of lignin content in Arabidopsis and the mature xylem layer of poplar. The presence of lignin-polysaccharides renders cell walls recalcitrant for pulping and biofuel production. The tissue-specific regulation of lignin biosynthesis, without altering all xylem in plants, can be utilized usefully by keeping mechanical strength and resistance to various environmental stimuli. To identify another pPLAIII homolog from Arabidopsis, constitutively overexpressed AtpPLAIIIα was characterized for xylem lignification in two well-studied model plants, Arabidopsis and poplar. The characterization of gene function in annual and perennial plants with respect to lignin biosynthesis revealed the functional redundancy of less lignification via downregulation of lignin biosynthesis-related genes.

scholarly journals Cell-wall polysaccharides and glycoproteins of parenchymatous tissues of runner bean (Phaseolus coccineus)

1990 ◽  
Vol 269 (2) ◽  
pp. 393-402 ◽  
Author(s):  
P Ryden ◽  
R R Selvendran
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Anion Exchange Chromatography ◽  
Structural Features ◽  
Good Evidence ◽  
Exchange Chromatography ◽  
Cell Wall Polysaccharides ◽  
Pectic Polysaccharides ◽  
Runner Bean ◽  
Cellulose Residue

1. Polymers were solubilized from the cell walls of parenchyma from mature runner-bean pods with minimum degradation by successive extractions with cyclohexane-trans-1,2-diamine-NNN′N′-tetra-acetate (CDTA), Na2CO3 and KOH to leave the alpha-cellulose residue, which contained cross-linked pectic polysaccharides and Hyp-rich glycoproteins. These were solubilized with chlorite/acetic acid and cellulase. The polymers were fractionated by anion-exchange chromatography, and fractions were subjected to methylation analysis. 2. The pectic polysaccharides differed in their ease of extraction, and a small proportion were highly cross-linked. The bulk of the pectic polysaccharides solubilized by CDTA and Na2CO3 were less branched than those solubilized by KOH. There was good evidence that most of the pectic polysaccharides were not degraded during extraction. 3. The protein-containing fractions included Hyp-rich and Hyp-poor glycoproteins associated with easily extractable pectic polysaccharides, Hyp-rich glycoproteins solubilized with 4M-KOH+borate, the bulk of which were not associated with pectic polysaccharides, and highly cross-linked Hyp-rich glycoproteins. 4. Isodityrosine was not detected, suggesting that it does not have a (major) cross-linking role in these walls. Instead, it is suggested that phenolics, presumably linked to C-5 of 3,5-linked Araf residues of Hyp-rich glycoproteins, serve to cross-link some of the polymers. 5. There were two main types of xyloglucan, with different degrees of branching. The bulk of the less branched xyloglucans were solubilized by more-concentrated alkali. The anomeric configurations of the sugars in one of the highly branched xyloglucans were determined by 13C-n.m.r. spectroscopy. 6. The structural features of the cell-wall polymers and complexes are discussed in relation to the structure of the cell walls of parenchyma tissues.

scholarly journals Kinetic properties and physiological role of the plastoquinone terminal oxidase (PTOX) in a vascular plant

2012 ◽  
Vol 1817 (12) ◽  
pp. 2140-2148 ◽  
Author(s):  
Martin Trouillard ◽  
Maryam Shahbazi ◽  
Lucas Moyet ◽  
Fabrice Rappaport ◽  
Pierre Joliot ◽  
...  
Keyword(s):  
Vascular Plant ◽  
Physiological Role ◽  
Kinetic Properties ◽  
Terminal Oxidase

scholarly journals Carbonic anhydrase isoenzymes in the erthrocytes and dorsolateral prostate of the rat

1969 ◽  
Vol 114 (3) ◽  
pp. 463-476 ◽  
Author(s):  
J. E. A. McIntosh
Keyword(s):  
Carbonic Anhydrase ◽  
High Activity ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Exchange Chromatography ◽  
Kinetic Properties ◽  
Efficient Catalyst ◽  
Molecular Weights ◽  
Hydrolysis Of ◽  
Low Activity

1. Three forms of the zinc-containing enzyme carbonic anhydrase (EC 4.2.1.1) were isolated from the erythrocytes of the rat and two forms from the dorsolateral prostate of the rat. Several additional minor components were observed but not isolated. Separation of the isoenzymes was achieved by ion-exchange chromatography, polyacrylamide-gel electrophoresis and isoelectric focusing. 2. The general properties of the isolated isoenzymes, their molecular weights and their contents of zinc were closely similar. As catalysts of the hydration of carbon dioxide, however, they were distinctly different. The two most abundant isoenzymes of the erythrocytes, which were found in equal proportions, differed 70-fold in specific activity, whereas the isoenzymes of the dorsolateral prostate were similar to one another and resembled the high-activity component of the erythrocytes. The inhibition of the latter by acetazolamide (5-acetamido-1-thia-3,4-diazole-2-sulphonamide) was mainly competitive, whereas in identical conditions the low-activity erythrocyte component and the dorsolateral prostate isoenzymes were non-competitively inhibited. 3. The use of chloroform–ethanol to remove haemoglobin from the rat haemolysate was found (a) to bring about changes in the kinetic properties of the soluble isoenzymes and (b) to cause the appearance of an additional isoenzyme. 4. The actions were compared of the inhibitors acetazolamide, 1,1-dimethylaminonaphthalene-5-sulphonamide and ethoxzolamide (6-ethoxybenzothiazole-2-sulphonamide) on the hydrolysis of p-nitrophenyl acetate catalysed by the isoenzymes. 5. The low-activity erythrocyte isoenzyme was an efficient catalyst of the hydrolysis of β-naphthyl acetate whereas the high-activity forms were much less active towards this ester. Neither of the isoenzymes present in the dorsolateral prostate catalysed this reaction. 6. Carbonic anhydrase in the rat dorsolateral prostate accounts for no more than 5% of the unusually high content of zinc in this organ.

scholarly journals A New Subfamily of Polyphosphate Kinase 2 (Class III PPK2) Catalyzes both Nucleoside Monophosphate Phosphorylation and Nucleoside Diphosphate Phosphorylation

2014 ◽  
Vol 80 (8) ◽  
pp. 2602-2608 ◽  
Author(s):  
Kei Motomura ◽  
Ryuichi Hirota ◽  
Mai Okada ◽  
Takeshi Ikeda ◽  
Takenori Ishida ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Amino Acid ◽  
High Energy ◽  
Class I ◽  
Polyphosphate Kinase ◽  
Kinetic Properties ◽  
Class Iii ◽  
Inorganic Polyphosphate ◽  
Pyrimidine Bases ◽  
Single Enzyme

ABSTRACTInorganic polyphosphate (polyP) is a linear polymer of tens to hundreds of phosphate (Pi) residues linked by “high-energy” phosphoanhydride bonds as in ATP. PolyP kinases, responsible for the synthesis and utilization of polyP, are divided into two families (PPK1 and PPK2) due to differences in amino acid sequence and kinetic properties. PPK2 catalyzes preferentially polyP-driven nucleotide phosphorylation (utilization of polyP), which is important for the survival of microbial cells under conditions of stress or pathogenesis. Phylogenetic analysis suggested that the PPK2 family could be divided into three subfamilies (classes I, II, and III). Class I and II PPK2s catalyze nucleoside diphosphate and nucleoside monophosphate phosphorylation, respectively. Here, we demonstrated that class III PPK2 catalyzes both nucleoside monophosphate and nucleoside diphosphate phosphorylation, thereby enabling us to synthesize ATP from AMP by a single enzyme. Moreover, class III PPK2 showed broad substrate specificity over purine and pyrimidine bases. This is the first demonstration that class III PPK2 possesses both class I and II activities.

scholarly journals Involvement of a Class III Peroxidase and the Mitochondrial Protein TSPO in Oxidative Burst Upon Treatment of Moss Plants with a Fungal Elicitor

2012 ◽  
Vol 25 (3) ◽  
pp. 363-371 ◽  
Author(s):  
Mikko T. Lehtonen ◽  
Motomu Akita ◽  
Wolfgang Frank ◽  
Ralf Reski ◽  
Jari P. T. Valkonen
Keyword(s):  
Oxidative Burst ◽  
Physcomitrella Patens ◽  
Redox Homeostasis ◽  
Mitochondrial Protein ◽  
Fungal Elicitor ◽  
Class Iii ◽  
Wild Type ◽  
Necrotrophic Pathogen ◽  
Free Heme ◽  
Class Iii Peroxidase

Production of apoplastic reactive oxygen species (ROS), or oxidative burst, is among the first responses of plants upon recognition of microorganisms. It requires peroxidase or NADPH oxidase (NOX) activity and factors maintaining cellular redox homeostasis. Here, PpTSPO1 involved in mitochondrial tetrapyrrole transport and abiotic (salt) stress tolerance was tested for its role in biotic stress in Physcomitrella patens, a nonvascular plant (moss). The fungal elicitor chitin caused an immediate oxidative burst in wild-type P. patens but not in the previously described ΔPrx34 mutants lacking the chitin-responsive secreted class III peroxidase (Prx34). Oxidative burst in P. patens was associated with induction of the oxidative stress-related genes AOX, LOX7, and NOX, and also PpTSPO1. The available ΔPpTSPO1 knockout mutants overexpressed AOX and LOX7 constitutively, produced 2.6-fold more ROS than wild-type P. patens, and exhibited increased sensitivity to a fungal necrotrophic pathogen and a saprophyte. These results indicate that Prx34, which is pivotal for antifungal resistance, catalyzes ROS production in P. patens, while PpTSPO1 controls redox homeostasis. The capacity of TSPO to bind harmful free heme and porphyrins and scavenge them through autophagy, as shown in Arabidopsis under abiotic stress, seems important to maintenance of the homeostasis required for efficient pathogen defense.

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