Characterisation of the Genes Encoding Mannuronan-C5-Epimerase in the Brown Alga Lessonia Variegata

<p>Alginate is known to be a commercially valuable polysaccharide, of great importance in industries such as food, cosmetics, medicine and pharmaceuticals. It is obtained commercially by harvesting brown algae. The final step in the alginate biochemical pathway involves the epimerization of D-mannuronic residues into L-guluronic residues, catalyzed by the enzyme mannuronan-C5-epimerase. This final step has been found to be responsible for controlling the physicochemical properties of the produced alginate. This study is the first to characterize the genes encoding for the enzyme mannuronan-C5- epimerase in the Northern, Southern and Wellington lineages of the brown alga Lessonia variegata (Phaeophyceae). The gene of interest was amplified by standard PCR and cloning. Cloning PCR results revealed the presence of two distinct copies of the gene in Lessonia variegata. The coding region of the copies was found to be very conserved with very little sequence variation. The Lessonia variegata sequences were compared with those of Laminaria digitata and Saccharina japonica, which indicated that at least one gene duplication event has occurred in Lessonia variegata, leading to the formation of two gene duplicates. The possible mechanisms by which the gene paralogs may control the structure and function of the produced alginate have been discussed.</p>

Characterisation of the Genes Encoding Mannuronan-C5-Epimerase in the Brown Alga Lessonia Variegata

<p>Alginate is known to be a commercially valuable polysaccharide, of great importance in industries such as food, cosmetics, medicine and pharmaceuticals. It is obtained commercially by harvesting brown algae. The final step in the alginate biochemical pathway involves the epimerization of D-mannuronic residues into L-guluronic residues, catalyzed by the enzyme mannuronan-C5-epimerase. This final step has been found to be responsible for controlling the physicochemical properties of the produced alginate. This study is the first to characterize the genes encoding for the enzyme mannuronan-C5- epimerase in the Northern, Southern and Wellington lineages of the brown alga Lessonia variegata (Phaeophyceae). The gene of interest was amplified by standard PCR and cloning. Cloning PCR results revealed the presence of two distinct copies of the gene in Lessonia variegata. The coding region of the copies was found to be very conserved with very little sequence variation. The Lessonia variegata sequences were compared with those of Laminaria digitata and Saccharina japonica, which indicated that at least one gene duplication event has occurred in Lessonia variegata, leading to the formation of two gene duplicates. The possible mechanisms by which the gene paralogs may control the structure and function of the produced alginate have been discussed.</p>

Trunk Surgery as a Tool to Reduce Foliar Symptoms in Diseases of the Esca Complex and Its Influence on Vine Wood Microbiota

In the last few years, trunk surgery has gained increasing attention as a method to reduce foliar symptoms typical of some of the Esca complex diseases. The technique relies on the mechanical removal of decayed wood by a chainsaw. A study on a 14-year-old Cabernet Sauvignon vineyard was carried out to validate the efficacy of trunk surgery and explore possible explanations behind it. Three levels of treatment were applied to three of the most characteristic symptoms associated with some diseases of the Esca complex, such as leaf stripe symptoms (LS), wilted shoots (WS) and apoplexy (APP). The most promising results were obtained by complete trunk surgery, where the larger decay removal allowed lower symptom re-expression. According to the wood types analyzed (decay, medium and sound wood), different changes in microbiota were observed. Alpha-diversity generally decreased for bacteria and increased for fungi. More specifically, main changes were observed for Fomitiporia mediterranea abundance that decreased considerably after trunk surgery. A possible explanation for LS symptom reduction after trunk surgery could be the microbiota shifting caused by the technique itself affecting a microbic-shared biochemical pathway involved in symptom expression.

Potential drug targets in the kynurenine pathway to treat acute schizophrenia

AimsSchizophrenia is a serious developmental psychiatric disorder with a neurodegenerative component that causes marked deterioration in social relationships and ability to work. Present treatments are not satisfactory. Meta-analysis of placebo-controlled studies in acute schizophrenia shows that only a minority of patients have a good response to current antipsychotic medications. Therefore, there is a need for more effective psychopharmacologic treatments for this disorder.MethodThe purpose of this paper is to provide new interpretations of existing data to provide a scaffolding for the development of novel drug targets for the treatment of schizophrenia. The causes of schizophrenia are most likely heterogeneous and involve both genetic and environmental factors. The authors examined a wide range of purported causes of schizophrenia to identify a common biochemical pathway that would contribute to this disorder. This review specifically did not consider pathways that supported the dopamine hypothesis of schizophrenia since historically drugs focused on dopaminergic mechanism, as noted in the aims, have not been successful for many patients with schizophrenia.ResultTwo prominent schizophrenia-associated factors that have been widely studied with significant supporting evidence are stress and inflammation. Stress and inflammation share a common biochemical pathway that converges on the kynurenine pathway of the metabolism of tryptophan, an essential amino acid. At one end of the pathway, recently hospitalized patients with schizophrenia have been found to have low plasma tryptophan levels, whereas chronic schizophrenics have not, suggesting stress- and/or inflammation-induced increased metabolism of tryptophan. At the other end of the pathway, there is increased level of cerebrospinal fluid kynurenic acid in patients with schizophrenia as compared with healthy controls. Salivary kynurenic acid is associated with stress intolerance in schizophrenia. Importantly, natural occurring compounds in this pathway have significant CNS effects that include neurotoxicity and altered neural transmitter behavior.ConclusionStress and inflammation, both associated as causes of schizophrenia, are linked by a common biochemical pathway involving kynurenine. Examination of specific elements of the kynurenine pathway may aid in the identification of drug targets for schizophrenia.

A New Pathway for Forming Acetate and Synthesizing ATP during Fermentation in Bacteria

Many bacteria and other organisms carry out fermentations forming acetate. These fermentations have broad importance to foods, agriculture, and industry. They also are important to bacteria themselves because they often generate ATP. Here we found a biochemical pathway for forming acetate and synthesizing ATP that was unknown in fermentative bacteria. We found the bacterium Cutibacterium granulosum formed acetate during fermentation of glucose. It did not use phosphotransacetylase or acetate kinase, enzymes found in nearly all acetate-forming bacteria. Instead, it used a pathway involving two different enzymes. The first enzyme, succinyl-CoA:acetate CoA-transferase (SCACT), forms acetate from acetyl-CoA. The second enzyme, succinyl-CoA synthetase (SCS), synthesizes ATP. We identified the genes encoding these enzymes, and they were homologs of SCACT and SCS genes found in other bacteria. The pathway resembles one described in eukaryotes, but it uses bacterial, not eukaryotic, gene homologs. To find other instances of the pathway, we analyzed sequences of all biochemically-characterized homologs of SCACT and SCS (103 enzymes from 64 publications). Homologs with similar enzymatic activity had similar sequences, enabling a large-scale search for them in genomes. We searched nearly 600 genomes of bacteria known to form acetate, and we found 6% encoded homologs with SCACT and SCS activity. This included >30 species belonging to 5 different phyla, showing a diverse range of bacteria encode the SCACT/SCS pathway. This work suggests the SCACT/SCS pathway is important to forming acetate in many branches of the tree of life. Importance Pathways for forming acetate during fermentation have been studied for over 80 years. In that time, several pathways have been described in a range of organisms, from bacteria to animals. However, one pathway (involving succinyl-CoA:acetate CoA-transferase and succinyl-CoA synthetase) has not been reported in prokaryotes. Here we discovered enzymes for this pathway in the fermentative bacterium Cutibacterium granulosum. We also found >30 other fermentative bacteria that encode this pathway, demonstrating it could be common. This pathway represents a new way for bacteria to form acetate from acetyl-CoA and synthesize ATP via substrate-level phosphorylation. It could be a target for controlling yield of acetate during fermentation, with relevance to foods, agriculture, and industry.

An ever-green mind and a heart for the colors of fall

With finest biochemical and molecular approaches, convincing explorative strategies and long-term vision, Stefan Hörtensteiner succeeded in elucidating the biochemical pathway responsible for chlorophyll degradation. After having contributed to the identification of key chlorophyll degradation products in the course of the last twenty-five years, he gradually identified and characterized most of the crucial players in the PAO/phyllobilin degradation pathway of chlorophyll. One of the brightest plant biochemists of his generation, his work opened doors to a better understanding of plant senescence, tetrapyrrole homeostasis and their complex regulation. He sadly passed away on 5 December 2020, aged 57.

Kearns Sayre Syndrome: A Mitochondrial Disorder

Kearns–Sayre syndrome (KSS) is a pleiotropic disorder caused by non-specific spontaneous deletion of a large amount of genetic material from mitochondrial DNA (mtDNA). Aside from patients having mtDNA defects there are also autosomal mutations in nuclear DNA, indicating KSS is not caused by a single gene mutation, but is most likely the result of mutations in genes forming a common biochemical pathway. KSS is characterized by a wide array of symptoms including: ophthalmoplegia, pigmentary retinopathy, ataxia, cardiac conduction defects that later develop into cardiac complications, and brain abnormalities. This review considers the association of deletions in mtDNA with a decrease in mitochondrial function and the pathogenetic role of the dysfunctional mitochondria by analyzing different variants of the mitochondrial genome. Despite there being no curative treatment for these patients, some possible disease modifying therapies have been proposed such as folinic acid supplementation and intravenous arginine therapy.

Promiscuous enzymes generating d-amino acids in mammals: Why they may still surprise us?

Promiscuous catalysis is a common property of enzymes, particularly those using pyridoxal 5′-phosphate as a cofactor. In a recent issue of this journal, Katane et al. Biochem. J. 477, 4221–4241 demonstrate the synthesis and accumulation of d-glutamate in mammalian cells by promiscuous catalysis mediated by a pyridoxal 5′-phosphate enzyme, the serine/threonine dehydratase-like (SDHL). The mechanism of SDHL resembles that of serine racemase, which synthesizes d-serine, a well-established signaling molecule in the mammalian brain. d-Glutamate is present in body fluids and is degraded by the d-glutamate cyclase at the mitochondria. This study demonstrates a biochemical pathway for d-glutamate synthesis in mammalian cells and advances our knowledge on this little-studied d-amino acid in mammals. d-Amino acids may still surprise us by their unique roles in biochemistry, intercellular signaling, and as potential biomarkers of disease.