scholarly journals Overexpression of nicotinamide mononucleotide adenyltransferase (nmnat) increases the growth rate, Ca2+ content and cellulase production in Ganoderma lucidum

2020 ◽  
Author(s):  
Shengli Wang ◽  
Jing Han ◽  
Jiale Xia ◽  
Yanru Hu ◽  
Liang Shi ◽  
...  
Keyword(s):  
Ganoderma Lucidum ◽  
Carbon Sources ◽  
Nutrient Utilization ◽  
Cellulase Production ◽  
Cellulosic Biomass ◽  
Gene Transcript ◽  
Transcript Levels ◽  
Nicotinamide Mononucleotide ◽  
Wide Range ◽  
Intracellular Ca

Abstract Background: There is an urgent need to identify new and economical ways to utilize diverse types of lignocellulosic biomass. Ganoderma lucidum is well-known edible medicinal fungus that has an excellent ability to degrade a wide range of cellulosic biomass, and its nutrient utilization is closely related to the production of extracellular cellulase. Nicotinamide adenine dinucleotide (NAD+), a nutritional sensor molecule, can respond to nutritional states and regulate cellular metabolism, and nicotinamide mononucleotide adenyltransferase (nmnat) is the key enzyme that catalyses the biosynthesis of NAD+.Results: In this study, a homologue of the gene encoding nmnat was cloned from G. lucidum. The Agrobacterium tumefaciens-mediated transformation (ATMT) method was used to construct two G. lucidum overexpression strains, OE::nmnat4 and OE::nmnat19, in which the nmnat gene transcript levels and the NAD+ content were significantly increased. Glnmnat overexpression strains showed dramatically increased colony growth on different carbon sources, and the intracellular Ca2+ concentration was increased by 3.95- and 2.10-fold in the G. lucidum OE::nmnat4 and OE::nmnat19 strains, respectively, compared with that observed in the WT strain. The CMCase activity increased by approximately 2.8- and 3-fold, and that of pNPGase increased by approximately 1.9- and 2.1-fold in the G. lucidum OE::nmnat4 and OE::nmnat19 strains, respectively, compared with that observed in the WT strain. Furthermore, NAD+ was observed to potentially induce cellulase production by regulating the cytosolic Ca2+ concentration.Conclusions: Taken together, for the first time, our results revealed that NAD+ can stimulate cellulase production and increase the transcript levels of cellulase genes via increasing the intracellular Ca2+ concentration in G. lucidum. This research also provides a theoretical basis for conducting cellulase-related work in other basidiomycetes.

scholarly journals Overexpression of nicotinamide mononucleotide adenyltransferase (nmnat) increases growth rate, Ca2+ content and cellulase production in Ganoderma lucidum

2019 ◽  
Author(s):  
shengli Wang ◽  
jing Han ◽  
Jiale Xia ◽  
Yanru Hu ◽  
Liang Shi ◽  
...  
Keyword(s):  
Ganoderma Lucidum ◽  
Carbon Sources ◽  
Nutrient Utilization ◽  
Colony Growth ◽  
Cellulase Production ◽  
Cellulosic Biomass ◽  
Transcript Levels ◽  
Medicinal Fungus ◽  
Nicotinamide Mononucleotide ◽  
Intracellular Ca

Abstract Background: There is an urgent need to search for new and economical ways to utilize diverse lignocellulose. Ganoderma lucidum is well-known edible medicinal fungus that has a strong ability to degrade a wide variety of cellulosic biomass and its nutrient utilization is closely related to extracellular cellulase. Nicotinamide adenine dinucleotide (NAD+), a nutritional sensor molecule, can respond to nutritional states and regulate cellular metabolism. Nicotinamide mononucleotide adenyltransferase (nmnat) is the key enzyme that catalyses the biosynthesis of NAD+.Result: In this study, a homologue of the gene encoding nmnat was cloned from G. lucidum. The Agrobacterium tumefaciens -mediated transformation (ATMT) method was used to construct the two overexpression strains OE:: nmnat4 and OE:: nmnat19 in G. lucidum. In the overexpression strains, the transcript levels of the nmnat gene and the NAD+ content were significantly increased. Gl nmnat overexpression strains showed dramatically stronger colony growth on different carbon sources, and the intracellular Ca 2+ concentration increased 3.95-fold and 2.10-fold in OE:: nmnat4 and OE:: nmnat19 , respectively, compared with WT strains. The CMCase activity increased by approximately 2.8-fold and 3-fold, and p NPGase activity increased by approximately 1.9-fold and 2.1-fold in OE:: nmnat4 and OE:: nmnat19 strains compared with the WT strains, respectively. Furthermore, it was found that NAD+ might induce cellulase production by regulating cytosolic Ca 2+ concentration.Conclusions: Taken together, our results revealed for the first time that NAD + could stimulate cellulase production and demonstrated that NAD + could increase the transcript levels of cellulase genes via the intracellular Ca 2+ concentration in G. lucidum. This research also provides a theoretical basis for conducting cellulase-related work on other basidiomycetes.

scholarly journals Burkholderia thailandensis E264 as a promising safe rhamnolipids’ producer towards a sustainable valorization of grape marcs and olive mill pomace

2021 ◽  
Author(s):  
Alif Chebbi ◽  
Massimiliano Tazzari ◽  
Cristiana Rizzi ◽  
Franco Hernan Gomez Tovar ◽  
Sara Villa ◽  
...  
Keyword(s):  
Energy Source ◽  
Olive Oil ◽  
Low Cost ◽  
Carbon Sources ◽  
Burkholderia Thailandensis ◽  
Rhamnolipid Production ◽  
Key Points ◽  
Wide Range ◽  
Long Chain ◽  
Olive Mill

Abstract Within the circular economy framework, our study aims to assess the rhamnolipid production from winery and olive oil residues as low-cost carbon sources by nonpathogenic strains. After evaluating various agricultural residues from those two sectors, Burkholderia thailandensis E264 was found to use the raw soluble fraction of nonfermented (white) grape marcs (NF), as the sole carbon and energy source, and simultaneously, reducing the surface tension to around 35 mN/m. Interestingly, this strain showed a rhamnolipid production up to 1070 mg/L (13.37 mg/g of NF), with a higher purity, on those grape marcs, predominately Rha-Rha C14-C14, in MSM medium. On olive oil residues, the rhamnolipid yield of using olive mill pomace (OMP) at 2% (w/v) was around 300 mg/L (15 mg/g of OMP) with a similar CMC of 500 mg/L. To the best of our knowledge, our study indicated for the first time that a nonpathogenic bacterium is able to produce long-chain rhamnolipids in MSM medium supplemented with winery residues, as sole carbon and energy source. Key points • Winery and olive oil residues are used for producing long-chain rhamnolipids (RLs). • Both higher RL yields and purity were obtained on nonfermented grape marcs as substrates. • Long-chain RLs revealed stabilities over a wide range of pH, temperatures, and salinities

scholarly journals Sporulation in solventogenic and acetogenic clostridia

2021 ◽  
Author(s):  
Mamou Diallo ◽  
Servé W. M. Kengen ◽  
Ana M. López-Contreras
Keyword(s):  
Current Knowledge ◽  
Carbon Sources ◽  
Cost Effective ◽  
Biotechnological Production ◽  
Key Points ◽  
Wide Range ◽  
Potential Applications ◽  
A Cell ◽  
Sporulation Initiation ◽  
Solventogenic Clostridia

AbstractThe Clostridium genus harbors compelling organisms for biotechnological production processes; while acetogenic clostridia can fix C1-compounds to produce acetate and ethanol, solventogenic clostridia can utilize a wide range of carbon sources to produce commercially valuable carboxylic acids, alcohols, and ketones by fermentation. Despite their potential, the conversion by these bacteria of carbohydrates or C1 compounds to alcohols is not cost-effective enough to result in economically viable processes. Engineering solventogenic clostridia by impairing sporulation is one of the investigated approaches to improve solvent productivity. Sporulation is a cell differentiation process triggered in bacteria in response to exposure to environmental stressors. The generated spores are metabolically inactive but resistant to harsh conditions (UV, chemicals, heat, oxygen). In Firmicutes, sporulation has been mainly studied in bacilli and pathogenic clostridia, and our knowledge of sporulation in solvent-producing or acetogenic clostridia is limited. Still, sporulation is an integral part of the cellular physiology of clostridia; thus, understanding the regulation of sporulation and its connection to solvent production may give clues to improve the performance of solventogenic clostridia. This review aims to provide an overview of the triggers, characteristics, and regulatory mechanism of sporulation in solventogenic clostridia. Those are further compared to the current knowledge on sporulation in the industrially relevant acetogenic clostridia. Finally, the potential applications of spores for process improvement are discussed.Key Points• The regulatory network governing sporulation initiation varies in solventogenic clostridia.• Media composition and cell density are the main triggers of sporulation.• Spores can be used to improve the fermentation process.

The vulnerability of Arctic shelf sediments to climate change

2015 ◽  
Vol 23 (4) ◽  
pp. 461-479 ◽  
Author(s):  
Robie W. Macdonald ◽  
Zou Zou A. Kuzyk ◽  
Sophia C. Johannessen
Keyword(s):  
Organic Carbon ◽  
Carbon Sources ◽  
The Arctic ◽  
Sedimentary Environments ◽  
Carbon Supply ◽  
Shelf Sediment ◽  
Wide Range ◽  
Metabolic Conditions ◽  
Shelf Sediments ◽  
Ocean Ecosystem

The sediments of the pan-Arctic shelves contribute an important component to the Arctic Ocean ecosystem by providing a habitat for biota (benthos), a repository for organic and inorganic non-conservative substances entering or produced within the ocean, a reactor and source of transformed substances back to the water column, and a mechanism of burial. Sediments interact with ice, ocean, and the surrounding land over a wide range of space and time scales. We discuss the vulnerability of shelf sediment to changes in (i) organic carbon sources, (ii) pathways of sediment and organic carbon supply, and (iii) physical and biogeochemical alteration (diagenesis). Sedimentary environments of the shelves and basins are likely to exhibit a wide variance in their response to global change because of their wide variation in sediment sources, processes, and metabolic conditions. In particular, the Chukchi and Barents shelves are dominated by inflowing waters from oceans to the south, whereas the interior shelves are more closely tied to terrigenous sources due to river inflow and coastal erosion.

scholarly journals Cross-generational feeding of Bt (Bacillus thuringiensis)-maize to zebrafish (Danio rerio) showed no adverse effects on the parental or offspring generations

2013 ◽  
Vol 110 (12) ◽  
pp. 2222-2233 ◽  
Author(s):  
Monica Sanden ◽  
Robin Ornsrud ◽  
Nini H. Sissener ◽  
Susanne Jorgensen ◽  
Jinni Gu ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Danio Rerio ◽  
Fumonisin B1 ◽  
Gene Transcript ◽  
Transcript Levels ◽  
Two Generations ◽  
Feed Utilisation ◽  
Liver Gene ◽  
Caspase 6 ◽  
Mass Increase

In the present study, zebrafish (Danio rerio) were fed casein/gelatin-based diets containing either 19 % Bt (Bacillus thuringiensis)-maize or its parental non-Bt (nBt)-maize control for two generations (F0: sixty fish; F1: forty-two to seventy fish per treatment). The study focused on growth and reproductive performance, liver CuZn superoxide dismutase (SOD) enzyme activity, gene transcript levels targeting important cellular pathways in the liver and mid-intestine, histomorphological evaluation of the intestine, differential leucocyte counts, offspring larva swimming activity and global DNA methylation in offspring embryos. No significant effects were observed in the parental generation. The offspring were either fed the same diets as those fed to their parents (Bt–Bt or nBt–nBt) or switched from the Bt diet to the nBt diet (Bt–nBt). The Bt–Bt offspring exhibited a significantly higher body mass increase, specific growth rate and feed utilisation than fish fed the nBt–nBt diet and/or fish fed the Bt–nBt diet. Liver and mid-intestinal gene transcript levels of CuZn SOD were significantly higher in fish fed the nBt–nBt diet than in those fed the Bt–Bt diet. Liver gene transcript levels of caspase 6 were significantly lower for the nBt–nBt group than for the Bt–Bt group. Overall, enhanced growth performance was observed in fish fed the Bt diet for two generations than in those fed the nBt diet for one and two generations. Effects observed on gene biomarkers for oxidative stress and the cell cycle (apoptosis) may be related to the contamination of nBt-maize with fumonisin B1 and aflatoxin B1. In conclusion, it is suggested that Bt-maize is as safe and nutritious as its nBt control when fed to zebrafish for two generations.

scholarly journals From Residues to Added-Value Bacterial Biopolymers as Nanomaterials for Biomedical Applications

Nanomaterials ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1492
Author(s):  
Francisco G. Blanco ◽  
Natalia Hernández ◽  
Virginia Rivero-Buceta ◽  
Beatriz Maestro ◽  
Jesús M. Sanz ◽  
...  
Keyword(s):  
Biomedical Applications ◽  
Carbon Sources ◽  
Structural Diversity ◽  
Added Value ◽  
Nanoscale Systems ◽  
Chemical Structures ◽  
Wide Range ◽  
Peptide Functionalization ◽  
Metabolic Strategies ◽  
Drug Nanocarriers

Bacterial biopolymers are naturally occurring materials comprising a wide range of molecules with diverse chemical structures that can be produced from renewable sources following the principles of the circular economy. Over the last decades, they have gained substantial interest in the biomedical field as drug nanocarriers, implantable material coatings, and tissue-regeneration scaffolds or membranes due to their inherent biocompatibility, biodegradability into nonhazardous disintegration products, and their mechanical properties, which are similar to those of human tissues. The present review focuses upon three technologically advanced bacterial biopolymers, namely, bacterial cellulose (BC), polyhydroxyalkanoates (PHA), and γ-polyglutamic acid (PGA), as models of different carbon-backbone structures (polysaccharides, polyesters, and polyamides) produced by bacteria that are suitable for biomedical applications in nanoscale systems. This selection models evidence of the wide versatility of microorganisms to generate biopolymers by diverse metabolic strategies. We highlight the suitability for applied sustainable bioprocesses for the production of BC, PHA, and PGA based on renewable carbon sources and the singularity of each process driven by bacterial machinery. The inherent properties of each polymer can be fine-tuned by means of chemical and biotechnological approaches, such as metabolic engineering and peptide functionalization, to further expand their structural diversity and their applicability as nanomaterials in biomedicine.

Abstract 15: Glucose-Mediated Remodeling of Cardiac DNA Methylation

2017 ◽  
Vol 121 (suppl_1) ◽  
Author(s):  
Mark E Pepin ◽  
David K Crossman ◽  
Joseph P Barchue ◽  
Salpy V Pamboukian ◽  
Steven M Pogwizd ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heart Failure ◽  
Dna Methylation ◽  
Diabetic Cardiomyopathy ◽  
Left Ventricular ◽  
Epigenetic Mark ◽  
Gene Transcript ◽  
Transcript Levels ◽  
Glycemic Memory

To identify the role of glucose in the development of diabetic cardiomyopathy, we had directly assessed glucose delivery to the intact heart on alterations of DNA methylation and gene expression using both an inducible heart-specific transgene (glucose transporter 4; mG4H) and streptozotocin-induced diabetes (STZ) mouse models. We aimed to determine whether long-lasting diabetic complications arise from prior transient exposure to hyperglycemia via a process termed “glycemic memory.” We had identified DNA methylation changes associated with significant gene expression regulation. Comparing our results from STZ, mG4H, and the modifications which persist following transgene silencing, we now provide evidence for cardiac DNA methylation as a persistent epigenetic mark contributing to glycemic memory. To begin to determine which changes contribute to human heart failure, we measured both RNA transcript levels and whole-genome DNA methylation in heart failure biopsy samples (n = 12) from male patients collected at left ventricular assist device placement using RNA-sequencing and Methylation450 assay, respectively. We hypothesized that epigenetic changes such as DNA methylation distinguish between heart failure etiologies. Our findings demonstrated that type 2 diabetic heart failure patients (n = 6) had an overall signature of hypomethylation, whereas patients listed as ischemic (n = 5) had a distinct hypermethylation signature for regulated transcripts. The focus of this initial analysis was on promoter-associated CpG islands with inverse changes in gene transcript levels, from which diabetes (14 genes; e.g. IGFBP4) and ischemic (12 genes; e.g. PFKFB3) specific targets emerged with significant regulation of both measures. By combining our mouse and human molecular analyses, we provide evidence that diabetes mellitus governs direct regulation of cellular function by DNA methylation and the corresponding gene expression in diabetic mouse and human hearts. Importantly, many of the changes seen in either mouse type 1 diabetes or human type 2 diabetes were similar supporting a consistent mechanism of regulation. These studies are some of the first steps at defining mechanisms of epigenetic regulation in diabetic cardiomyopathy.

Relationship between flow rate and NO production in postnatal mesenteric arteries

2001 ◽  
Vol 280 (1) ◽  
pp. G43-G50 ◽  
Author(s):  
Kristina M. Reber ◽  
Gennifer M. Mager ◽  
Charles E. Miller ◽  
Philip T. Nowicki
Keyword(s):  
Flow Rate ◽  
Kinase Inhibitor ◽  
Age Groups ◽  
Mesenteric Arteries ◽  
No Production ◽  
Herbimycin A ◽  
Wide Range ◽  
Intracellular Ca ◽  
Controlled Flow

We studied mesenteric arterial arcades from 3- and 35-day-old swine to determine the relationship between perfusate flow rate and release of nitric oxide (NO) into mesenteric effluent. Mesenteric arterial arcades were perfused under controlled-flow conditions with a peristaltic pump using warm oxygenated Krebs buffer. Basal rates of NO production were 43.6 ± 4.2 vs. 12.1 ± 2.5 nmol/min in 3- vs. 35-day-old mesentery during perfusion at in vivo flow rates (9 vs. 20 ml/min, respectively). Rate of NO production was directly related to flow rate over a wide range of flows (5–40 ml/min) in 3- but not 35-day-old mesentery. Both age groups demonstrated a brisk, albeit brief, increase in NO production in response to infusion of NO-dependent vasodilator substance P (10−8 M/min). Tyrosine kinase inhibitor herbimycin A andl-arginine analog l-NMMA significantly attenuated flow-induced increase in NO production, and phosphatase inhibitor phenylarsine oxide increased magnitude of flow-induced increase in NO production in 3-day-olds. Removal of extracellular Ca2+ and depletion of intracellular Ca2+ stores (Ca2+-free Krebs with EGTA plus thapsigargin) had no effect on NO production in either group. Thus, basal rate of NO production is greater in mesenteric arterial arcades from 3- than from 35-day old swine, a direct relationship between flow rate and NO production rate is present in mesentery from 3- but not 35-day-olds, and phosphorylation events are necessary for this interaction to occur.

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