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 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.

Functions of reactive oxygen species in apoptosis and ganoderic acid biosynthesis in Ganoderma lucidum

2019 ◽  
Vol 366 (23) ◽  
Author(s):  
Jing Zhu ◽  
Fengli Wu ◽  
Sining Yue ◽  
Chen Chen ◽  
Shuqi Song ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Ganoderma Lucidum ◽  
Reactive Oxygen ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Activity Levels ◽  
Oxygen Species ◽  
Ganoderic Acid ◽  
Medicinal Fungus ◽  
Intracellular Ros ◽  
Ga Biosynthesis

ABSTRACT Ganoderma lucidum is a medicinal fungus that is widely used in traditional medicine. Fungal PacC is recognized as an important transcription factor that functions during adaptation to environmental pH, fungal development and secondary metabolism. Previous studies have revealed that GlPacC plays important roles in mycelial growth, fruiting body development and ganoderic acid (GA) biosynthesis. In this study, using a terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay, we found that the apoptosis level was increased when PacC was silenced. The transcript and activity levels of caspase-like proteins were significantly increased in the PacC-silenced (PacCi) strains compared with the control strains. Silencing PacC also resulted in an increased reactive oxygen species (ROS) levels (∼2-fold) and decreased activity levels of enzymes involved in the antioxidant system. Further, we found that the intracellular ROS levels contributed to apoptosis and GA biosynthesis. Adding N-acetyl-cysteine and vitamin C decreased intracellular ROS and resulted in the inhibition of apoptosis in the PacCi strains. Additionally, the GA biosynthesis was different between the control strains and the PacCi strains after intracellular ROS was eliminated. Taken together, the findings showed that silencing PacC can result in an intracellular ROS burst, which increases cell apoptosis and GA biosynthesis levels. Our study provides novel insight into the functions of PacC in filamentous fungi.

scholarly journals ACEI of Trichoderma reesei Is a Repressor of Cellulase and Xylanase Expression

2003 ◽  
Vol 69 (1) ◽  
pp. 56-65 ◽  
Author(s):  
Nina Aro ◽  
Marja Ilmén ◽  
Anu Saloheimo ◽  
Merja Penttilä
Keyword(s):  
Trichoderma Reesei ◽  
Culture Media ◽  
Carbon Sources ◽  
Cellulase Production ◽  
The Novel ◽  
Hypocrea Jecorina ◽  
Gene Encoding ◽  
Gene Coding ◽  
Better Than

ABSTRACT We characterized the effect of deletion of the Trichoderma reesei (Hypocrea jecorina) ace1 gene encoding the novel cellulase regulator ACEI that was isolated based on its ability to bind to and activate in vivo in Saccharomyces cerevisiae the promoter of the main cellulase gene, cbh1. Deletion of ace1 resulted in an increase in the expression of all the main cellulase genes and two xylanase genes in sophorose- and cellulose-induced cultures, indicating that ACEI acts as a repressor of cellulase and xylanase expression. Growth of the strain with a deletion of the ace1 gene on different carbon sources was analyzed. On cellulose-based medium, on which cellulases are needed for growth, the Δace1 strain grew better than the host strain due to the increased cellulase production. On culture media containing sorbitol as the sole carbon source, the growth of the strain with a deletion of the ace1 gene was severely impaired, suggesting that ACEI regulates expression of other genes in addition to cellulase and xylanase genes. A strain with a deletion of the ace1 gene and with a deletion of the ace2 gene coding for the cellulase and xylanase activator ACEII expressed cellulases and xylanases similar to the Δace1 strain, indicating that yet another activator regulating cellulase and xylanase promoters was present.

scholarly journals Genetic and Metabolic Intraspecific Biodiversity ofGanoderma lucidum

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Anna Pawlik ◽  
Grzegorz Janusz ◽  
Iwona Dębska ◽  
Marek Siwulski ◽  
Magdalena Frąc ◽  
...  
Keyword(s):  
Ganoderma Lucidum ◽  
Mycelial Growth ◽  
Carbon Sources ◽  
Fungal Growth ◽  
Its Region ◽  
Mitochondrial Activity ◽  
Genomic Relationship ◽  
Profile Similarity ◽  
Geographic Regions ◽  
Its Region Sequencing

FourteenGanoderma lucidumstrains from different geographic regions were identified using ITS region sequencing. Based on the sequences obtained, the genomic relationship between the analyzed strains was determined. AllG. lucidumstrains were also genetically characterized using the AFLP technique.G. lucidumstrains included in the analysis displayed an AFLP profile similarity level in the range from 9.6 to 33.9%. Biolog FF MicroPlates were applied to obtain data on utilization of 95 carbon sources and mitochondrial activity. The analysis allowed comparison of functional diversity of the fungal strains. The substrate utilization profiles for the isolates tested revealed a broad variability within the analyzedG. lucidumspecies and proved to be a good profiling technology for studying the diversity in fungi. Significant differences have been demonstrated in substrate richness values. Interestingly, the analysis of growth and biomass production also differentiated the strains based on the growth rate on the agar and sawdust substrate. In general, the mycelial growth on the sawdust substrate was more balanced and the fastest fungal growth was observed for GRE3 and FCL192.

Ganoderma lucidum Polysaccharide Strengthens Antioxidant Defense Systems against Exercise-Induced Oxidative Stress in Liver of Mice

2014 ◽  
Vol 675-677 ◽  
pp. 1577-1579
Author(s):  
Lin Luo ◽  
Feng Yan ◽  
Bei Bei Wang
Keyword(s):  
Oxidative Stress ◽  
Chinese Medicine ◽  
Traditional Chinese Medicine ◽  
Ganoderma Lucidum ◽  
Antioxidant Defense ◽  
Defense Systems ◽  
Medicinal Fungus ◽  
Traditional Remedy ◽  
Antioxidant Defense Systems ◽  
Exercise Induced

Ganoderma lucidum(G. lucidum), a medicinal fungus called ‘‘Lingzhi’’ in China is one of the most famous fungi in traditional Chinese medicine. For hundreds of years, this mushroom has been known as a traditional remedy for treatment of several diseases, such as hepatitis, hypertension, chronic bronchitis, bronchial asthma, cancer and others. Polysaccharide is an important pharmacological ingredient extracted from fruit bodies and mycelium ofG. lucidum. It has been extensively documented thatG. lucidumpolysaccharides (Gl-PS) had wide bioactivities, such as anti-aging, anti-tumor, hypoglycemic, immunomodulatory and immunotherapeutic activities [1]. In our previous studies, Gl-PS has been proven to have significant anti-hypoxia and anti-fatigue effects in mice [2], which suggesting that it might be beneficial to exercise-induced oxidative stress. Therefore, the purpose of this study was to investigate the effects of Gl-PS against exercise-induced oxidative stress in liver of mice.

scholarly journals The Carbon Source Controls the Secretion and Yield of Polysaccharide-Hydrolyzing Enzymes of Basidiomycetes

2021 ◽  
Author(s):  
Eka Metreveli ◽  
Tamar Khardziani ◽  
Vladimir Elisashvili
Keyword(s):  
Carbon Source ◽  
Carbon Sources ◽  
Schizophyllum Commune ◽  
Cellulase Production ◽  
Crystalline Cellulose ◽  
Irpex Lacteus ◽  
Xylanase Production ◽  
Hydrolyzing Enzymes ◽  
Pycnoporus Coccineus ◽  
Polymeric Substrates

In the present study, the polysaccharide-hydrolyzing secretomes of Irpex lacteus BCC104, Pycnoporus coccineus BCC310, and Schizophyllum commune BCC632 were analyzed in submerged fermentation conditions to elucidate the effect of chemically and structurally different carbon sources on the expression of cellulases and xylanase. Among polymeric substrates, crystalline cellulose appeared to be the best carbon source providing the highest endoglucanase, total cellulase, and xylanase activities. Mandarin pomace as a growth substrate for S. commune allowed to achieve comparatively high volumetric activities of all target enzymes while wheat straw induced a significant secretion of cellulase and xylanase activities of I. lacteus and P. coccineus. A synergistic effect on the secretion of cellulases and xylanases by the tested fungi was observed when crystalline cellulose was combined with mandarin pomace. In I. lacteus the cellulase and xylanase production is inducible in the presence of cellulose-rich substrates but is suppressed in the presence of an excess of easily metabolizable carbon source. These enzymes are expressed in a coordinated manner under all conditions studied. It was shown that the substitution of glucose in the inoculum medium with Avicel provides accelerated enzyme production by I. lacteus and higher cellulase and xylanase activities of the fungus. These results add new knowledge to the physiology of basidiomycetes to improve cellulase production.

Sign in / Sign up

Export Citation Format

Share Document