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Keywords = glycerol-3-phosphate dehydrogenase genes

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15 pages, 3853 KiB  
Article
Enhanced Stress Tolerance in Rice Through Overexpression of a Chimeric Glycerol-3-Phosphate Dehydrogenase (OEGD)
by Jinhong Wu, Meiyao Chen, Fangwen Yang, Jing Han, Xiaosong Ma, Tianfei Li, Hongyan Liu, Bin Liang and Shunwu Yu
Plants 2025, 14(11), 1731; https://doi.org/10.3390/plants14111731 - 5 Jun 2025
Cited by 1 | Viewed by 473
Abstract
Crop productivity is severely constrained by abiotic and biotic stresses, necessitating innovative strategies to enhance stress resilience. Glycerol-3-phosphate (G3P) is a central metabolite in carbohydrate and lipid metabolism, playing crucial roles in stress responses. In this study, we engineered a novel glycerol-3-phosphate dehydrogenase [...] Read more.
Crop productivity is severely constrained by abiotic and biotic stresses, necessitating innovative strategies to enhance stress resilience. Glycerol-3-phosphate (G3P) is a central metabolite in carbohydrate and lipid metabolism, playing crucial roles in stress responses. In this study, we engineered a novel glycerol-3-phosphate dehydrogenase (GPDH) gene, designated OEGD, by fusing the N-terminal NAD-binding domain of rice OsGPDH1 with the feedback-resistant C-terminal catalytic domain of Escherichia coli gpsA. Overexpression of OEGD in rice enhanced tolerance to drought, phosphorus deficiency, high temperature, and cadmium (Cd2+) stresses, while also improving plant growth and yield under drought stress at the adult stage. Notably, the accumulation of glycerol-3-phosphate (G3P) and activities of antioxidant enzymes (SOD, POD, CAT) were significantly elevated in the transgenic plants following osmotic stimuli, and fatty acid profiles were altered, favoring stress adaptation. Transcriptomic analyses revealed that OEGD modulates cell wall biogenesis, reactive oxygen species (ROS) scavenging, and lipid metabolism pathways, with minimal disruption to core G3P metabolic genes. These findings highlight the potential of OEGD as a valuable genetic resource for improving stress resistance in rice. Full article
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17 pages, 6119 KiB  
Article
Phosphorus Functional Genes Control Rice Yield via Microbial Biomass Phosphorus and Plant Phosphorus Uptake in a Rice–Oilseed Rape Rotation System Compared with a Rice–Wheat Rotation System
by Qingyue Zhang, Weijia Yu, Min Li, Wenlong Cheng, Shengchang Huai, Yuwen Jin, Guihua Li, Ji Wu and Changai Lu
Agronomy 2025, 15(4), 866; https://doi.org/10.3390/agronomy15040866 - 30 Mar 2025
Viewed by 1394
Abstract
Crop rotation and microbial driving force significantly influence soil phosphorus (P) bioavailability and crop yield. However, differences in underlying microbial mechanisms in rotations remain unclear. We examined rice yield, P uptake, soil and microbial P contents, enzyme activity, and P functional genes over [...] Read more.
Crop rotation and microbial driving force significantly influence soil phosphorus (P) bioavailability and crop yield. However, differences in underlying microbial mechanisms in rotations remain unclear. We examined rice yield, P uptake, soil and microbial P contents, enzyme activity, and P functional genes over six years (2016–2022) to elucidate microbial mechanisms driving rice yield in rice–wheat (RW) and rice–oilseed rape (RO) rotations. RO significantly increased rice yield and plant P uptake by 9.17% and 20.70%, respectively, compared to RW. Soil total (TP) and available (AP) P contents were significantly lower (4.83% and 18.31%, respectively) under RO than RW, whereas microbial biomass phosphorus (MBP) and acid phosphatase activity (EP) were greater (39.40% and 128.45%, respectively). PICRUSt2 results revealed that RO increased phoA phoB (alkaline phosphatase), phnX (phosphonoacetaldehyde hydrolase [EC:3.11.1.1]), gcd (Quinoprotein glucose dehydrogenase [EC:1.1.5.2]), and ppaC (manganese-dependent inorganic pyrophosphatase) and decreased phnD (phosphonate transport system substrate-binding protein), ugpE (sn-glycerol 3-phosphate transport system permease protein), ugpA (sn-glycerol 3-phosphate transport system permease protein), and phnO ((aminoalkyl)phosphonate N-acetyltransferase [EC:2.3.1.280]) abundance. Random forest analysis showed that ppaC, phnD, gcd, and phnX were important for rice yield and plant P uptake. Partial least squares analysis revealed that RO indirectly increased rice yield by influencing MBP and affecting plant P uptake through P functional genes. Overall, RO improves rice yield and P bioavailability by altering P functional genes (ppaC, phnD, gcd, and phnX), providing new perspectives on crop–microorganism interactions and resource use efficiency. Full article
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15 pages, 5395 KiB  
Article
Transcriptome and Expression Analysis of Glycerol Biosynthesis-Related Genes in Glenea cantor Fabricius (Cerambycidae: Lamiinae)
by Taihui Lan, Ranran Su, Zishu Dong, Xin Tong, Xialin Zheng and Xiaoyun Wang
Int. J. Mol. Sci. 2024, 25(21), 11834; https://doi.org/10.3390/ijms252111834 - 4 Nov 2024
Viewed by 1064
Abstract
Glenea cantor Fabricius (Cerambycidae: Lamiinae) is an important pest that damages kapok trees in Southeast Asia with a wide adaptability to temperature. Glycerol is a protectant and energy source for insects in low-temperature environments. However, glycerol biosynthesis-related genes at the molecular level are [...] Read more.
Glenea cantor Fabricius (Cerambycidae: Lamiinae) is an important pest that damages kapok trees in Southeast Asia with a wide adaptability to temperature. Glycerol is a protectant and energy source for insects in low-temperature environments. However, glycerol biosynthesis-related genes at the molecular level are limited in G. cantor. In this study, the supercooling points and freezing points at different stages were measured, and the cold hardiness of male and female pupae significantly differed. Moreover, a full-length transcriptome of G. cantor was established; glycerol kinase (GK) and glycerol-3-phosphate dehydrogenase (GPDH) genes, which are related to glycerol metabolism, were identified, with a special focus on their expression profiles. A total of 24,476 isoforms stemmed from the full-length transcriptome, along with 568 lncRNAs, 56 transcription factor (TF) families, and 1467 alternative splicing (AS) events. The KEGG pathway enrichment analysis revealed that the isoforms associated with AS were enriched primarily in glycerolipid and glycerophospholipid metabolism. In total, three GK genes and one GPDH gene were identified, and GcGK1 and GcGK3 presented differential sex expression during the pupal stage, which may play a role in thermal adaptability. This study provides a valuable transcriptional database of G. cantor and helps to elucidate the function of glycerol in the thermal adaptation mechanism of longhorn beetles. Full article
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13 pages, 663 KiB  
Article
Meat Characteristics, Expression of Myosin Heavy Chain and Metabolism-Related Genes in Thai Native Pigs
by Chanporn Chaosap, Kamon Chaweewan, Kazeem D. Adeyemi, Netanong Phonkate and Ronachai Sitthigripong
Foods 2024, 13(10), 1502; https://doi.org/10.3390/foods13101502 - 13 May 2024
Viewed by 1521
Abstract
This study investigated the meat quality, expression of myosin heavy chain (MyHC) and metabolism-related genes, ribonucleotides and fatty acids in Longissimus thoracis of Thai native pigs (TNPs) from different geographical regions (GR). Forty-one 9–10-month-old castrated TNPs (BW 60 kg), consisting of 18, 11 [...] Read more.
This study investigated the meat quality, expression of myosin heavy chain (MyHC) and metabolism-related genes, ribonucleotides and fatty acids in Longissimus thoracis of Thai native pigs (TNPs) from different geographical regions (GR). Forty-one 9–10-month-old castrated TNPs (BW 60 kg), consisting of 18, 11 and 12 pigs from Northern (NT), Southern (ST) and Northeastern (NE) regions, respectively, were slaughtered. GR did not affect (p > 0.05) the expression of MyHC, phosphoglycerate mutase 1, cytosolic glycerol-3-phosphate dehydrogenase, triosephosphate isomerase 1 and adipocyte fatty acid binding protein genes. The trend of MyHC was MyHC IIx > MyHC IIb > MyHC IIa > MyHC I. The NT loin had higher (p < 0.05) glycogen, C18:2n6, C20:4n6 and cooking loss, lower inosine, inosine monophosphate and hypoxanthine and a shorter sarcomere length than the ST and NE loins. The ST loin had a lower (p < 0.05) a* compared to other loins. Principal component analysis established significant relationships between the TNP and specific meat quality traits. This finding suggests that GR affected the meat quality, ribonucleotides and selected fatty acids in TNPs. These results provide relevant information that can be used to optimize the use of Thai native pork. Full article
(This article belongs to the Section Meat)
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23 pages, 4911 KiB  
Article
Aspergillus nidulans gfdB, Encoding the Hyperosmotic Stress Protein Glycerol-3-phosphate Dehydrogenase, Disrupts Osmoadaptation in Aspergillus wentii
by Veronika Bodnár, Károly Antal, Ronald P. de Vries, István Pócsi and Tamás Emri
J. Fungi 2024, 10(4), 291; https://doi.org/10.3390/jof10040291 - 16 Apr 2024
Cited by 1 | Viewed by 2034
Abstract
The genome of the osmophilic Aspergillus wentii, unlike that of the osmotolerant Aspergillus nidulans, contains only the gfdA, but not the gfdB, glycerol 3-phosphate dehydrogenase gene. Here, we studied transcriptomic changes of A. nidulans (reference strain and ΔgfdB [...] Read more.
The genome of the osmophilic Aspergillus wentii, unlike that of the osmotolerant Aspergillus nidulans, contains only the gfdA, but not the gfdB, glycerol 3-phosphate dehydrogenase gene. Here, we studied transcriptomic changes of A. nidulans (reference strain and ΔgfdB gene deletion mutant) and A. wentii (reference strain and An-gfdB expressing mutant) elicited by high osmolarity. A. nidulans showed a canonic hyperosmotic stress response characterized by the upregulation of the trehalose and glycerol metabolism genes (including gfdB), as well as the genes of the high-osmolarity glycerol (HOG) map kinase pathway. The deletion of gfdB caused only negligible alterations in the transcriptome, suggesting that the glycerol metabolism was flexible enough to compensate for the missing GfdB activity in this species. A. wentii responded differently to increased osmolarity than did A. nidulans, e.g., the bulk upregulation of the glycerol and trehalose metabolism genes, along with the HOG pathway genes, was not detected. The expression of An-gfdB in A. wentii did not abolish osmophily, but it reduced growth and caused much bigger alterations in the transcriptome than did the missing gfdB gene in A. nidulans. Flexible glycerol metabolism and hence, two differently regulated gfd genes, may be more beneficial for osmotolerant (living under changing osmolarity) than for osmophilic (living under constantly high osmolarity) species. Full article
(This article belongs to the Special Issue Stress Research in Filamentous Fungi and Yeasts)
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13 pages, 1454 KiB  
Article
Jasmonates and Ethylene Shape Floridoside Synthesis during Carposporogenesis in the Red Seaweed Grateloupia imbricata
by Pilar Garcia-Jimenez, Diana del Rosario-Santana and Rafael R. Robaina
Mar. Drugs 2024, 22(3), 115; https://doi.org/10.3390/md22030115 - 28 Feb 2024
Cited by 2 | Viewed by 2344
Abstract
Floridoside is a galactosyl–glycerol compound that acts to supply UDP-galactose and functions as an organic osmolyte in response to salinity in Rhodophyta. Significantly, the UDP-galactose pool is shared for sulfated cell wall galactan synthesis, and, in turn, affected by thallus development alongside carposporogenesis [...] Read more.
Floridoside is a galactosyl–glycerol compound that acts to supply UDP-galactose and functions as an organic osmolyte in response to salinity in Rhodophyta. Significantly, the UDP-galactose pool is shared for sulfated cell wall galactan synthesis, and, in turn, affected by thallus development alongside carposporogenesis induced by volatile growth regulators, such as ethylene and methyl jasmonate, in the red seaweed Grateloupia imbricata. In this study, we monitored changes in the floridoside reservoir through gene expression controlling both the galactose pool and glyceride pool under different reproductive stages of G. imbricata and we considered changing salinity conditions. Floridoside synthesis was followed by expression analysis of galactose-1-phosphate uridyltransferase (GALT) as UDP-galactose is obtained from UDP-glucose and glucose-1P, and through α-galactosidase gene expression as degradation of floridoside occurs through the cleavage of galactosyl residues. Meanwhile, glycerol 3-phosphate is connected with the galactoglyceride biosynthetic pathway by glycerol 3-phosphate dehydrogenase (G3PD), monogalactosyl diacylglyceride synthase (MGDGS), and digalactosyl diacylglyceride synthase (DGDGS). The results of our study confirm that low GALT transcripts are correlated with thalli softness to locate reproductive structures, as well as constricting the synthesis of UDP-hexoses for galactan backbone synthesis in the presence of two volatile regulators and methionine. Meanwhile, α-galactosidase modulates expression according to cystocarp maturation, and we found high transcripts in late development stages, as occurred in the presence of methyljasmonate, compared to early stages in ethylene. Regarding the acylglyceride pool, the upregulation of G3PD, MGDGS, and DGDGS gene expression in G. imbricata treated with MEJA supports lipid remodeling, as high levels of transcripts for MGDGS and DGDGS provide membrane stability during late development stages of cystocarps. Similar behavior is assumed in three naturally collected thalli development stages—namely, fertile, fertilized, and fertile—under 65 psu salinity conditions. Low transcripts for α-galactosidase and high for G3PD are reported in infertile and fertilized thalli, which is the opposite to high transcripts for α-galactosidase and low for G3PD encountered in fertile thalli within visible cystocarps compared to each of their corresponding stages in 35 psu. No significant changes are reported for MGDGS and DGDGS. It is concluded that cystocarp and thallus development stages affect galactose and glycerides pools with interwoven effects on cell wall polysaccharides. Full article
(This article belongs to the Special Issue Characterization of Bioactive Components in Edible Algae 3rd Edition)
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13 pages, 1586 KiB  
Article
Increasing the Pentose Phosphate Pathway Flux to Improve Plasmid DNA Production in Engineered E. coli
by Mitzi de la Cruz, Flavio Kunert, Hilal Taymaz-Nikerel, Juan-Carlos Sigala, Guillermo Gosset, Jochen Büchs and Alvaro R. Lara
Microorganisms 2024, 12(1), 150; https://doi.org/10.3390/microorganisms12010150 - 12 Jan 2024
Viewed by 3056
Abstract
The demand of plasmid DNA (pDNA) as a key element for gene therapy products, as well as mRNA and DNA vaccines, is increasing together with the need for more efficient production processes. An engineered E. coli strain lacking the phosphotransferase system and the [...] Read more.
The demand of plasmid DNA (pDNA) as a key element for gene therapy products, as well as mRNA and DNA vaccines, is increasing together with the need for more efficient production processes. An engineered E. coli strain lacking the phosphotransferase system and the pyruvate kinase A gene has been shown to produce more pDNA than its parental strain. With the aim of improving pDNA production in the engineered strain, several strategies to increase the flux to the pentose phosphate pathway (PPP) were evaluated. The simultaneous consumption of glucose and glycerol was a simple way to increase the growth rate, pDNA production rate, and supercoiled fraction (SCF). The overexpression of key genes from the PPP also improved pDNA production in glucose, but not in mixtures of glucose and glycerol. Particularly, the gene coding for the glucose 6-phosphate dehydrogenase (G6PDH) strongly improved the SCF, growth rate, and pDNA production rate. A linear relationship between the G6PDH activity and pDNA yield was found. A higher flux through the PPP was confirmed by flux balance analysis, which also estimates relevant differences in fluxes of the tricarboxylic acid cycle. These results are useful for developing further cell engineering strategies to increase pDNA production and quality. Full article
(This article belongs to the Special Issue Advances in Microbial Cell Factories)
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19 pages, 3005 KiB  
Article
Morphological and Phylogenetic Analyses Reveal Three New Species of Phyllosticta (Botryosphaeriales, Phyllostictaceae) in China
by Yang Jiang, Zhaoxue Zhang, Jie Zhang, Shi Wang and Xiuguo Zhang
J. Fungi 2024, 10(1), 7; https://doi.org/10.3390/jof10010007 - 22 Dec 2023
Cited by 6 | Viewed by 2732
Abstract
The genus Phyllosticta has been reported worldwide and contains many pathogenic and endophytic species isolated from a wide range of plant hosts. A multipoint phylogeny based on gene coding combinatorial data sets for the internal transcribed spacer (ITS), large subunit of ribosomal RNA [...] Read more.
The genus Phyllosticta has been reported worldwide and contains many pathogenic and endophytic species isolated from a wide range of plant hosts. A multipoint phylogeny based on gene coding combinatorial data sets for the internal transcribed spacer (ITS), large subunit of ribosomal RNA (LSU rDNA), translation elongation factor 1α (TEF1α), actin (ACT), and glycerol-3-phosphate dehydrogenase (GPDH), combined with morphological characteristics, was performed. We describe three new species, P. fujianensis sp. nov., P. saprophytica sp. nov., and P. turpiniae sp. nov., and annotate and discusse their similarities and differences in morphological relationships and phylogenetic phases with closely related species. Full article
(This article belongs to the Special Issue Plant Pathogenic Fungi: Taxonomy, Phylogeny and Morphology)
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14 pages, 2282 KiB  
Article
Unraveling the Genetic Basis of Feed Efficiency in Cattle through Integrated DNA Methylation and CattleGTEx Analysis
by Zhenbin Hu, Clarissa Boschiero, Cong-Jun Li, Erin E. Connor, Ransom L. Baldwin and George E. Liu
Genes 2023, 14(12), 2121; https://doi.org/10.3390/genes14122121 - 24 Nov 2023
Cited by 6 | Viewed by 2664
Abstract
Feed costs can amount to 75 percent of the total overhead cost of raising cows for milk production. Meanwhile, the livestock industry is considered a significant contributor to global climate change due to the production of greenhouse gas emissions, such as methane. Indeed, [...] Read more.
Feed costs can amount to 75 percent of the total overhead cost of raising cows for milk production. Meanwhile, the livestock industry is considered a significant contributor to global climate change due to the production of greenhouse gas emissions, such as methane. Indeed, the genetic basis of feed efficiency (FE) is of great interest to the animal research community. Here, we explore the epigenetic basis of FE to provide base knowledge for the development of genomic tools to improve FE in cattle. The methylation level of 37,554 CpG sites was quantified using a mammalian methylation array (HorvathMammalMethylChip40) for 48 Holstein cows with extreme residual feed intake (RFI). We identified 421 CpG sites related to 287 genes that were associated with RFI, several of which were previously associated with feeding or digestion issues. Activator of transcription and developmental regulation (AUTS2) is associated with digestive disorders in humans, while glycerol-3-phosphate dehydrogenase 2 (GPD2) encodes a protein on the inner mitochondrial membrane, which can regulate glucose utilization and fatty acid and triglyceride synthesis. The extensive expression and co-expression of these genes across diverse tissues indicate the complex regulation of FE in cattle. Our study provides insight into the epigenetic basis of RFI and gene targets to improve FE in dairy cattle. Full article
(This article belongs to the Section Epigenomics)
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12 pages, 1657 KiB  
Article
Molecular Characterization of Ciborinia camelliae Kohn Shows Intraspecific Variability and Suggests Transcontinental Movement of the Pathogen
by Marco Saracchi, Irene Valenti, Paolo Cortesi, Daniela Bulgari, Andrea Kunova and Matias Pasquali
Microorganisms 2023, 11(11), 2727; https://doi.org/10.3390/microorganisms11112727 - 8 Nov 2023
Viewed by 1433
Abstract
Ciborinia camelliae Kohn is the causal agent of camellia flower blight. The fungus infects only the flowers of camellias. C. camelliae isolates obtained from symptomatic samples, collected in 13 different localities worldwide, were characterized by Multi-Locus Sequence Typing (MLST) using the following: (i) [...] Read more.
Ciborinia camelliae Kohn is the causal agent of camellia flower blight. The fungus infects only the flowers of camellias. C. camelliae isolates obtained from symptomatic samples, collected in 13 different localities worldwide, were characterized by Multi-Locus Sequence Typing (MLST) using the following: (i) a nuclear ribosomal DNA internal transcribed spacer; (ii) subunit 2 of β-tubulin (β-TUB II), (iii) elongation factor 1-α (EF1α); and (iv) glycerol-3-phosphate dehydrogenase (GPDH). The variability of the strains was assessed using a universally primed–polymerase chain reaction (UP-PCR) with six universal primers. Gene sequence comparison showed high similarity among all the European strains and highlighted the diversity of the New Zealand and Chinese representative strains. The profiles obtained by UP-PCR confirmed the significant diversity of extra-European strains and identified subgroups within the European population. The presence of shared genetic profiles obtained from strains isolated in different countries (New Zealand and France) suggests the movement of strains from one location to another, which is probably due to the exchange of infected plant material. Moreover, our study shows the overall high intraspecific variability of C. camelliae, which is likely due to the sexual reproduction of the fungus, suggesting the risk of emergence of new pathotypes adapting to novel camellia varieties. Full article
(This article belongs to the Special Issue Plant-Pathogenic Fungi)
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13 pages, 3345 KiB  
Article
Prognostic and Predictive Utility of GPD1L in Human Hepatocellular Carcinoma
by Philip K. H. Leung, Bibek Das, Xiaoyu Cheng and Munir Tarazi
Int. J. Mol. Sci. 2023, 24(17), 13113; https://doi.org/10.3390/ijms241713113 - 23 Aug 2023
Cited by 5 | Viewed by 2480
Abstract
Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths worldwide. GPD1L, a member of the glycerol-3-phosphate dehydrogenase family, has emerged as a potential tumour suppressor gene, with high expression associated with a favourable prognosis in various cancers. Despite an intriguing inverse relationship [...] Read more.
Hepatocellular carcinoma (HCC) is a major cause of cancer-related deaths worldwide. GPD1L, a member of the glycerol-3-phosphate dehydrogenase family, has emerged as a potential tumour suppressor gene, with high expression associated with a favourable prognosis in various cancers. Despite an intriguing inverse relationship observed with HCC, the precise role and underlying function of GPD1L in HCC remain poorly understood. Here, we aimed to investigate the prognostic significance, molecular characteristics, and predictive potential of GPD1L overexpression in HCC. Analysis of independent datasets revealed a significant correlation between high GPD1L expression and poor survival in HCC patients. Spatial and single cell transcriptome datasets confirmed elevated GDP1L expression in tumour tissue compared to adjacent normal tissue. GPD1L exhibited increased expression and promoter demethylation with advancing tumour stage, confirming positive selection during tumorigeneses. GPD1L overexpression was associated with metabolic dysregulation and enrichment of gene sets related to cell cycle control, epithelial-mesenchymal transition, and E2F targets. Moreover, we demonstrated an inverse correlation between GPD1L expression and therapeutic response for three therapeutic agents (PF-562271, Linsitinib, and BMS-754807), highlighting its potential as a predictive biomarker for HCC treatment outcomes. These data provide insights into the prognostic significance, molecular characteristics, and predictive potential of GPD1L in HCC. Full article
(This article belongs to the Special Issue Molecular Mechanisms of Liver Cancer)
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20 pages, 3034 KiB  
Article
Membrane Phenotypic, Metabolic and Genotypic Adaptations of Streptococcus oralis Strains Destined to Rapidly Develop Stable, High-Level Daptomycin Resistance during Daptomycin Exposures
by Nagendra N. Mishra, Rodrigo de Paula Baptista, Truc T. Tran, Christian K. Lapitan, Cristina Garcia-de-la-Maria, Jose M. Miró, Richard A. Proctor and Arnold S. Bayer
Antibiotics 2023, 12(7), 1083; https://doi.org/10.3390/antibiotics12071083 - 21 Jun 2023
Viewed by 1938
Abstract
The Streptococcus mitis-oralis subgroup of viridans group streptococci are important human pathogens. We previously showed that a substantial portion of S. mitis-oralis strains (>25%) are ‘destined’ to develop rapid, high-level, and stable daptomycin (DAP) resistance (DAP-R) during DAP exposures in vitro. Such DAP-R [...] Read more.
The Streptococcus mitis-oralis subgroup of viridans group streptococci are important human pathogens. We previously showed that a substantial portion of S. mitis-oralis strains (>25%) are ‘destined’ to develop rapid, high-level, and stable daptomycin (DAP) resistance (DAP-R) during DAP exposures in vitro. Such DAP-R is often accompanied by perturbations in distinct membrane phenotypes and metabolic pathways. The current study evaluated two S. oralis bloodstream isolates, 73 and 205. Strain 73 developed stable, high-level DAP-R (minimum inhibitory concentration [MIC] > 256 µg/mL) within 2 days of in vitro DAP passage (“high level” DAP-R [HLDR]). In contrast, strain 205 evolved low-level and unstable DAP-R (MIC = 8 µg/mL) under the same exposure conditions in vitro (“non-HLDR”). Comparing the parental 73 vs. 73-D2 (HLDR) strain-pair, we observed the 73-D2 had the following major differences: (i) altered cell membrane (CM) phospholipid profiles, featuring the disappearance of phosphatidylglycerol (PG) and cardiolipin (CL), with accumulation of the PG-CL pathway precursor, phosphatidic acid (PA); (ii) enhanced CM fluidity; (iii) increased DAP surface binding; (iv) reduced growth rates; (v) decreased glucose utilization and lactate accumulation; and (vi) increased enzymatic activity within the glycolytic (i.e., lactate dehydrogenase [LDH]) and lipid biosynthetic (glycerol-3-phosphate dehydrogenase [GPDH]) pathways. In contrast, the 205 (non-HLDR) strain-pair did not show these same phenotypic or metabolic changes over the 2-day DAP exposure. WGS analyses confirmed the presence of mutations in genes involved in the above glycolytic and phospholipid biosynthetic pathways in the 73-D2 passage variant. These data suggest that S. oralis strains which are ‘destined’ to rapidly develop HLDR do so via a conserved cadre of genotypic, membrane phenotypic, and metabolic adaptations. Full article
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14 pages, 1789 KiB  
Article
D-Lactic Acid Production from Sugarcane Bagasse by Genetically Engineered Saccharomyces cerevisiae
by Warasirin Sornlek, Kittapong Sae-Tang, Akaraphol Watcharawipas, Sriwan Wongwisansri, Sutipa Tanapongpipat, Lily Eurwilaichtr, Verawat Champreda, Weerawat Runguphan, Peter J. Schaap and Vitor A. P. Martins dos Santos
J. Fungi 2022, 8(8), 816; https://doi.org/10.3390/jof8080816 - 3 Aug 2022
Cited by 16 | Viewed by 3581
Abstract
Lactic acid (LA) is a promising bio-based chemical that has broad applications in food, nutraceutical, and bioplastic industries. However, production of the D-form of LA (D-LA) from fermentative organisms is lacking. In this study, Saccharomyces cerevisiae harboring the D-lactate dehydrogenase (DLDH) gene from [...] Read more.
Lactic acid (LA) is a promising bio-based chemical that has broad applications in food, nutraceutical, and bioplastic industries. However, production of the D-form of LA (D-LA) from fermentative organisms is lacking. In this study, Saccharomyces cerevisiae harboring the D-lactate dehydrogenase (DLDH) gene from Leuconostoc mesenteroides was constructed (CEN.PK2_DLDH). To increase D-LA production, the CRISPR/Cas12a system was used for the deletion of gpd1, gpd2, and adh1 to minimize glycerol and ethanol production. Although an improved D-LA titer was observed for both CEN.PK2_DLDHΔgpd and CEN.PK2_DLDHΔgpdΔadh1, growth impairment was observed. To enhance the D-LA productivity, CEN.PK2_DLDHΔgpd was crossed with the weak acid-tolerant S. cerevisiae BCC39850. The isolated hybrid2 showed a maximum D-LA concentration of 23.41 ± 1.65 g/L, equivalent to the improvement in productivity and yield by 2.2 and 1.5 folds, respectively. The simultaneous saccharification and fermentation using alkaline pretreated sugarcane bagasse by the hybrid2 led to an improved D-LA conversion yield on both the washed solid and whole slurry (0.33 and 0.24 g/g glucan). Our findings show the exploitation of natural yeast diversity and the potential strategy of gene editing combined with conventional breeding on improving the performance of S. cerevisiae for the production of industrially potent products. Full article
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25 pages, 3962 KiB  
Article
Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in Clostridium beijerinckii NCIMB 8052
by Eric Agyeman-Duah, Santosh Kumar, Bhavana Gangwar and Victor C. Ujor
Fermentation 2022, 8(7), 339; https://doi.org/10.3390/fermentation8070339 - 19 Jul 2022
Cited by 8 | Viewed by 3641
Abstract
Efficient bioconversion of abundant waste glycerol to value-added chemicals calls for a wider range of fermentative workhorses that can catabolize glycerol. In this study, we used quantitative gene expression and solvent profiling, qualitative metabolite analysis, and enzyme activity assays to investigate the factors [...] Read more.
Efficient bioconversion of abundant waste glycerol to value-added chemicals calls for a wider range of fermentative workhorses that can catabolize glycerol. In this study, we used quantitative gene expression and solvent profiling, qualitative metabolite analysis, and enzyme activity assays to investigate the factors that limit glycerol utilization as a sole carbon source by Clostridium beijerinckii NCIMB 8052. C. beijerinckii NCIMB 8052 did not produce acetate, acetone and butanol on glycerol. Congruently, the genes encoding the coenzyme A transferase subunits (ctfAB) and bifunctional acetaldehyde-CoA/alcohol dehydrogenase (adhE) were down-regulated up to 135- and 21-fold, respectively, at 12 h in glycerol-grown cells compared to glucose-grown cells. Conversely, NADH-dependent butanol dehydrogenase A (bdhA) was upregulated 2-fold. Glycerol dehydrogenase (gldA) and dihydroxyacetone kinase (subunit dhaK) were upregulated up to 5- and 881-fold, respectively. Glyceraldehyde-3-phosphate dehydrogenase (gapdh) showed mostly similar expression profiles at 12 h on glucose and glycerol. At 24 h, gapdh was downregulated 1.5-fold, while NADP+-dependent gapdh was upregulated up to 1.9-fold. Glycerol-grown cells showed higher or similar activity profiles for all solventogenic enzymes studied, compared to glucose-grown cells. Butyraldehyde (3 g/L) supplementation led to the production of ~0.1 g/L butanol, whilst butyrate (3.5 g/L) supplementation produced 0.7 and 0.5 g/L acetone and butanol, respectively, with glycerol. Further, the long chain saturated fatty acids cyclopentaneundecanoic acid, methyl ester and hexadecanoic acid, butyl ester were detected in glucose- but not in glycerol-grown cells. Collectively, growth on glycerol appears to disrupt synthesis of saturated long chain fatty acids, as well as solventogenesis in C. beijerinckii NCIMB 8052. Full article
(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)
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19 pages, 12239 KiB  
Article
Genome-Wide Identification of Cotton (Gossypium spp.) Glycerol-3-Phosphate Dehydrogenase (GPDH) Family Members and the Role of GhGPDH5 in Response to Drought Stress
by Jialiang Sun, Hua Cui, Bingjie Wu, Weipeng Wang, Qiuyue Yang, Yaxin Zhang, Song Yang, Yuping Zhao, Dongbei Xu, Guoxiang Liu and Tengfei Qin
Plants 2022, 11(5), 592; https://doi.org/10.3390/plants11050592 - 22 Feb 2022
Cited by 6 | Viewed by 3176
Abstract
Glycerol-3-phosphate dehydrogenase (GPDH) is a key enzyme in plant glycerol synthesis and metabolism, and plays an important role in plant resistance to abiotic stress. Here, we identified 6, 7, 14 and 14 GPDH genes derived from Gossypium arboreum, Gossypium raimondii, Gossypium [...] Read more.
Glycerol-3-phosphate dehydrogenase (GPDH) is a key enzyme in plant glycerol synthesis and metabolism, and plays an important role in plant resistance to abiotic stress. Here, we identified 6, 7, 14 and 14 GPDH genes derived from Gossypium arboreum, Gossypium raimondii, Gossypium barbadense and Gossypium hirsutum, respectively. Phylogenetic analysis assigned these genes into three classes, and most of the genes within the family were expanded by whole-genome duplication (WGD) and segmental duplications. Moreover, determination of the nonsynonymous substitution rate/synonymous substitution rate (Ka/Ks) ratio showed that the GPDH had an evolutionary preference for purifying selection. Transcriptome data revealed that GPDH genes were more active in the early stages of fiber development. Additionally, numerous stress-related cis-elements were identified in the potential promoter region. Then, a protein–protein-interaction (PPI) network of GPDH5 in G. hirsutum was constructed. In addition, we predicted 30 underlying miRNAs in G. hirsutum. Functional validation results indicated that silencing GhGPDH5 diminished drought tolerance in the upland cotton TM-1 line. In summary, this study provides a fundamental understanding of the GPDH gene family in cotton, GhGPDH5 exerts a positive effect during drought stress and is potentially involved in stomatal closure movements. Full article
(This article belongs to the Special Issue Molecular Breeding and Genetic Improvement of Cotton)
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