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1.
Sci Rep ; 14(1): 17481, 2024 07 30.
Artículo en Inglés | MEDLINE | ID: mdl-39080323

RESUMEN

Carbimazole has disadvantages on different body organs, especially the thyroid gland and, rarely, the adrenal glands. Most studies have not suggested any solution or medication for ameliorating the noxious effects of drugs on the glands. Our study focused on the production of xylooligosaccharide (XOS), which, when coadministered with carbimazole, relieves the toxic effects of the drug on the adrenal glands. In addition to accelerating the regeneration of adrenal gland cells, XOS significantly decreases the oxidative stress caused by obesity. This XOS produced by Aspergillus terreus xylanase was covalently immobilized using microbial Scleroglucan gel beads, which improved the immobilization yield, efficiency, and operational stability. Over a wide pH range (6-7.5), the covalent immobilization of xylanase on scleroglucan increased xylanase activity compared to that of its free form. Additionally, the reaction temperature was increased to 65 °C. However, the immobilized enzyme demonstrated superior thermal stability, sustaining 80.22% of its original activity at 60 °C for 120 min. Additionally, the full activity of the immobilized enzyme was sustained after 12 consecutive cycles, and the activity reached 78.33% after 18 cycles. After 41 days of storage at 4 °C, the immobilized enzyme was still active at approximately 98%. The immobilized enzyme has the capability to produce xylo-oligosaccharides (XOSs). Subsequently, these XOSs can be coadministered alongside carbimazole to mitigate the adverse effects of the drug on the adrenal glands. In addition to accelerating the regeneration of adrenal gland cells, XOS significantly decreases the oxidative stress caused by obesity.


Asunto(s)
Glándulas Suprarrenales , Aspergillus , Carbimazol , Enzimas Inmovilizadas , Oligosacáridos , Aspergillus/efectos de los fármacos , Oligosacáridos/farmacología , Oligosacáridos/química , Enzimas Inmovilizadas/metabolismo , Enzimas Inmovilizadas/química , Glándulas Suprarrenales/efectos de los fármacos , Glándulas Suprarrenales/metabolismo , Animales , Glucuronatos/farmacología , Estrés Oxidativo/efectos de los fármacos , Endo-1,4-beta Xilanasas/metabolismo , Masculino , Ratas , Obesidad/tratamiento farmacológico
2.
Carbohydr Polym ; 341: 122320, 2024 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-38876713

RESUMEN

In an oil exploitation process, hydrogel plugging agents can effectively reduce the water-oil intermixing, decrease water extraction volume, and enhance oil recovery rate. The practical applications of traditional polyacrylamide (PAM) hydrogel plugging agents in oilfield are limited by their non-biodegradability, poor mechanical performance, and inferior temperature-resistance. This work developed a mechanically stable and high-temperature-resistant composite hydrogel (STP) by incorporating biodegradable scleroglucan (Slg) and TEMPO-oxidized cellulose nanofibers (TOCN) in the PAM hydrogel. The addition of Slg conferred heat resistance to the PAM hydrogel, while TOCN reinforced the mechanical strength. Anti-aging analyses revealed that the STP endured for 108 h in a saline environment at 140 °C. In the water flooding characterization, the STP displayed a breakthrough pressure of 42.10 psi/ft. at a flow rate of 0.75 cm3/min. Under these extreme conditions, the plugging pressure reached 14.74 psi/ft., meeting the essential criteria for oilfield water plugging. This research demonstrates the potential of polysaccharides in the preparation of sustainable, tough, and heat-resistant water plugging materials.

3.
Int J Biol Macromol ; 272(Pt 2): 132864, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38844272

RESUMEN

Scleroglucan is a notable member of the ß-glucan microbial polysaccharides with a long tradition of industrial and therapeutic use. The local strain, previously identified as Athelia rolfsii TEMG MH 236106 produced an appreciable amount of scleroglucan using glucose as a carbon source and yeast extract as a nitrogen source. Plackett-Burman design was employed to effectively screen critical medium composition, culture, and fermentation conditions. Athelia rolfsii TEMG MH 236106 produced the maximum amount of scleroglucan (18.12 g/L) with a 45.3 % glucose conversion. Out of the eleven variables, the most effective factors showing a high level of significance are as follows: glucose, yeast extract, citric acid, inoculum disc numbers, culture volume and incubation time. An update to maximize scleroglucan production in the central composite design for four parameters (glucose and yeast extract concentrations, disc number, medium volume and incubation time) with 31 runs was applied and the production of scleroglucan reached its maximum at 31.56 g/L with 78.9 % glucose conversion. Three models of Sclg-5-fluorouracil complexes have been employed to study in vitro drug release investigations. Hence, the Sclg-5-FU (5 and 10 mg/mL) models appeared to be the most suitable for drug administration due to their concentration and distribution within capsules.


Asunto(s)
Fluorouracilo , Glucanos , Glucanos/química , Fluorouracilo/farmacología , Fluorouracilo/metabolismo , Fermentación , Liberación de Fármacos , Glucosa/metabolismo , Medios de Cultivo/química
4.
Nanomaterials (Basel) ; 14(8)2024 Apr 13.
Artículo en Inglés | MEDLINE | ID: mdl-38668170

RESUMEN

Scleroglucan (SG) is resistant to harsh reservoir conditions such as high temperature, high shear stresses, and the presence of chemical substances. However, it is susceptible to biological degradation because bacteria use SG as a source of energy and carbon. All degradation effects lead to viscosity loss of the SG solutions, affecting their performance as an enhanced oil recovery (EOR) polymer. Recent studies have shown that nanoparticles (NPs) can mitigate these degradative effects. For this reason, the EOR performance of two new nanohybrids (NH-A and NH-B) based on carboxymethyl-scleroglucan and amino-functionalized silica nanoparticles was studied. The susceptibility of these products to chemical, mechanical, and thermal degradation was evaluated following standard procedures (API RP 63), and the microbial degradation was assessed under reservoir-relevant conditions (1311 ppm and 100 °C) using a bottle test system. The results showed that the chemical reactions for the nanohybrids obtained modified the SG triple helix configuration, impacting its viscosifying power. However, the nanohybrid solutions retained their viscosity during thermal, mechanical, and chemical degradation experiments due to the formation of a tridimensional network between the nanoparticles (NPs) and the SG. Also, NH-A and NH-B solutions exhibited bacterial control because of steric hindrances caused by nanoparticle modifications to SG. This prevents extracellular glucanases from recognizing the site of catalysis, limiting free glucose availability and generating cell death due to substrate depletion. This study provides insights into the performance of these nanohybrids and promotes their application in reservoirs with harsh conditions.

5.
Nanomaterials (Basel) ; 14(6)2024 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-38535647

RESUMEN

In this study, two new nanohybrids (NH-A and NH-B) were synthesized through carbodiimide-assisted coupling. The reaction was performed between carboxymethyl-scleroglucans (CMS-A and CMS-B) with different degrees of substitution and commercial amino-functionalized silica nanoparticles using 4-(dimethylamino)-pyridine (DMAP) and N,N'-dicyclohexylcarbodiimide (DCC) as catalysts. The morphology and properties of the nanohybrids were investigated by using transmission (TEM) and scanning electron microscopy (SEM), electron-dispersive scanning (EDS), attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FT-IR), X-ray photoelectron spectroscopy (XPS), powder X-ray diffraction (XRD), inductively coupled plasma atomic emission spectroscopy (ICP-OES), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and dynamic light scattering (DLS). The nanohybrids exhibited differences in structure due to the incorporation of polyhedral oligomeric silsesquioxane (POSS) materials. The results reveal that hybrid nanomaterials exhibit similar thermal properties but differ in morphology, chemical structure, and crystallinity properties. Finally, a viscosity study was performed on the newly obtained nanohybrid materials; viscosities of nanohybrids increased significantly in comparison to the carboxymethyl-scleroglucans, with a viscosity difference of 7.2% for NH-A and up to 32.6% for NH-B.

6.
Int Immunopharmacol ; 129: 111614, 2024 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-38350358

RESUMEN

BACKGROUND: Intestinal tissue is extremely sensitive to ionizing radiation (IR), which is easy to cause intestinal radiation sickness, and the mortality rate is very high after exposure. Recent studies have found that intestinal immune cells and intestinal stem cells (ISCs) may play a key role in IR-induced intestinal injury. METHODS: C57BL6 mice matched for age, sex and weight were randomly grouped and intraperitoneal injected with PBS, Scleroglucan (125.0 mg/kg) or Anti-mouse IL-17A -InVivo (10 mg/kg), the number of mice in each group was n ≥ 3.Survival time, body weight, pathology, organoids and immune cell markers of the mice after IR (10.0 Gy) were compared, and the mechanism of action in intestinal tissues was verified by transcriptome sequencing. RESULTS: Scleroglucan has significant radiation protective effects on the intestine, including improving the survival rate of irradiated mice, inhibiting the radiation damage of intestinal tissue, and promoting the proliferation and differentiation of intestinal stem cells (ISCs). The results of RNA sequencing suggested that Scleroglucan could significantly activate the immune system and up-regulate the IL-17 and NF-κB signaling pathways. Flow cytometry showed that Scleroglucan could significantly up-regulate the number of Th17 cells and the level of IL-17A in the gut. IL-17A provides radiation protection. After intraperitoneal injection of Scleroglucan and Anti-mouse IL-17A -InVivo, mice can significantly reverse the radiation protection effect of Scleroglucan, down-regulate the molecular markers of intestinal stem cells and the associated markers of DC, Th1 and Th17 cells, and up-regulate the associated markers of Treg and Macrophage cells. CONCLUSION: Scleroglucan may promote the proliferation and regeneration of ISCs by regulating the activation of intestinal immune function mediated by IL-17 signaling pathway and play a protective role in IR-induced injury.


Asunto(s)
Glucanos , Traumatismos por Radiación , Protectores contra Radiación , Ratones , Animales , Interleucina-17 , Ratones Endogámicos C57BL , Traumatismos por Radiación/prevención & control , Transducción de Señal , Protectores contra Radiación/farmacología , Protectores contra Radiación/uso terapéutico , Intestinos/patología
7.
Nanomaterials (Basel) ; 14(2)2024 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-38251121

RESUMEN

Biopolymers emerge as promising candidates for enhanced oil recovery (EOR) applications due to their molecular structures, which exhibit better stability than polyacrylamides under harsh conditions. Nonetheless, biopolymers are susceptible to oxidation and biological degradation. Biopolymers reinforced with nanoparticles could be a potential solution to the issue. The nanofluids' stability and performance depend on the nanoparticles' properties and the preparation method. The primary objective of this study was to evaluate the effect of the preparation method and the nanoparticle type (SiO2, Al2O3, and TiO2) on the viscosity and stability of the scleroglucan (SG). The thickening effect of the SG solution was improved by adding all NPs due to the formation of three-dimensional structures between the NPs and the SG chains. The stability test showed that the SG + Al2O3 and SG + TiO2 nanofluids are highly unstable, but the SG + SiO2 nanofluids are highly stable (regardless of the preparation method). According to the ANOVA results, the preparation method and standing time influence the nanofluid viscosity with a statistical significance of 95%. On the contrary, the heating temperature and NP type are insignificant. Finally, the nanofluid with the best performance was 1000 ppm of SG + 100 ppm of SiO2_120 NPs prepared by method II.

8.
Polymers (Basel) ; 16(2)2024 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-38257006

RESUMEN

This paper presents the methodology for synthesizing and characterizing two carboxymethyl EOR-grade Scleroglucans (CMS-A and CMS-B). An O-Alkylation reaction was used to insert a hydrophilic group (monochloroacetic acid-MCAA) into the biopolymer's anhydroglucose subunits (AGUs). The effect of the degree of the carboxymethyl substitution on the rheology and thermal stability of the Scleroglucan (SG) was also evaluated. Simultaneous thermal analysis (STA/TGA-DSC), differential scanning calorimetry (DSC), X-ray Diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, Scanning Electron Microscopy, and Energy Dispersive Spectroscopy (SEM/EDS) were employed to characterize both CMS products. FTIR analysis revealed characteristic peaks corresponding to the carboxymethyl functional groups, confirming the modification. Also, SEM analysis provided insights into the structural changes in the polysaccharide after the O-Alkylation reaction. TGA results showed that the carboxymethylation of SG lowered its dehydroxylation temperature but increased its thermal stability above 300 °C. The CMS products and SG exhibited a pseudoplastic behavior; however, lower shear viscosities and relaxation times were observed for the CMS products due to the breakage of the SG triple helix for the chemical modification. Despite the viscosity results, the modified Scleroglucans are promising candidates for developing new engineering materials for EOR processes.

9.
Polymers (Basel) ; 15(21)2023 Nov 06.
Artículo en Inglés | MEDLINE | ID: mdl-37960018

RESUMEN

The chemically synthesized polymer polyacrylamide (HPAM) has achieved excellent oil displacement in conventional reservoirs, but its oil displacement is poor in extreme reservoir environments. To develop a biopolymer oil flooding agent suitable for extreme reservoir conditions, the viscosity changes and rheological properties of three biopolymers, diutan gum, xanthan gum, and scleroglucan, were studied under extreme reservoir conditions (high salt, high temperature, strong acid, and alkali), and the effects of temperature, mineralization, pH, and other factors on their viscosities and long-term stability were analyzed and compared. The results show that the three biopolymers had the best viscosity-increasing ability at temperatures of 90 °C and below. The viscosity of the three biopolymers was 80.94 mPa·s, 11.57 mPa·s, and 59.83 mPa·s, respectively, when the concentration was 1500 mg/L and the salinity 220 g/L. At the shear rate of 250 s-1, 100 °C~140 °C, scleroglucan had the best viscosification. At 140 °C, the solution viscosity was 19.74 mPa·s, and the retention rate could reach 118.27%. The results of the long-term stability study showed that the solution viscosity of scleroglucan with a mineralization level of 220 mg/L was 89.54% viscosity retention in 40 days, and the diutan gum could be stabilized for 10 days, with the viscosity maintained at 90 mPa·s. All three biopolymers were highly acid- and alkali-resistant, with viscosity variations of less than 15% in the pH3~10 range. Rheological tests showed that the unique double-helix structure of diutan gum and the rigid triple-helix structure of scleroglucan caused them to have better viscoelastic properties than xanthan gum. Therefore, these two biopolymers, diutan gum, and scleroglucan, have the potential for extreme reservoir oil displacement applications. It is recommended to use diutan gum for oil displacement in reservoirs up to 90 °C and scleroglucan for oil displacement in reservoirs between 100 °C and 140 °C.

10.
Cell Rep Med ; 4(5): 101024, 2023 05 16.
Artículo en Inglés | MEDLINE | ID: mdl-37119814

RESUMEN

RNA viruses continue to remain a threat for potential pandemics due to their rapid evolution. Potentiating host antiviral pathways to prevent or limit viral infections is a promising strategy. Thus, by testing a library of innate immune agonists targeting pathogen recognition receptors, we observe that Toll-like receptor 3 (TLR3), stimulator of interferon genes (STING), TLR8, and Dectin-1 ligands inhibit arboviruses, Chikungunya virus (CHIKV), West Nile virus, and Zika virus to varying degrees. STING agonists (cAIMP, diABZI, and 2',3'-cGAMP) and Dectin-1 agonist scleroglucan demonstrate the most potent, broad-spectrum antiviral function. Furthermore, STING agonists inhibit severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and enterovirus-D68 (EV-D68) infection in cardiomyocytes. Transcriptome analysis reveals that cAIMP treatment rescue cells from CHIKV-induced dysregulation of cell repair, immune, and metabolic pathways. In addition, cAIMP provides protection against CHIKV in a chronic CHIKV-arthritis mouse model. Our study describes innate immune signaling circuits crucial for RNA virus replication and identifies broad-spectrum antivirals effective against multiple families of pandemic potential RNA viruses.


Asunto(s)
COVID-19 , Virus Chikungunya , Virus ARN , Infección por el Virus Zika , Virus Zika , Animales , Ratones , SARS-CoV-2 , Antivirales/farmacología , Antivirales/uso terapéutico , Virus Chikungunya/fisiología , Inmunidad Innata
11.
Food Chem ; 404(Pt A): 134508, 2023 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-36219965

RESUMEN

The aim of this study was to determine the applicability of commonly used dyes (CWS and CFB) to stain different types of ß-glucans in water and selected, most popular, dairy products for CLSM imaging analysis. Structurally different pure ß-glucan preparations: curdlan-CU, oat ß-glucan-OG and scleroglucan-SC were tested. Our study showed for the first time that CWS is a dye with a specific affinity for all analyzed ß-glucans, while CFB is suitable for curdlan labeling only. Despite the technological processes, ß-glucans structures in dairy products are similar to those in aqueous suspension forms; each of the ß-glucans forms a different and characteristic structure, ranging from spindle-shaped and branched till granular. The presented results for the first time systematize the knowledge on CWS and CFB applicability in ß-glucan CLSM staining and allow future researchers to correctly analyze simultaneously stained ß-glucan, fat, and protein in a complex matrix as dairy products.


Asunto(s)
Agua , beta-Glucanos , beta-Glucanos/química , Colorantes Fluorescentes , Productos Lácteos , Rayos Láser
12.
World J Microbiol Biotechnol ; 39(2): 49, 2022 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-36542187

RESUMEN

Microbial exopolysaccharides (EPSs) are mostly produced by bacteria and fungi and have potential use in the production of biomedical products such as nutraceuticals and in tissue engineering applications. The present study investigated the in vitro biological activities and in vivo wound healing effects of EPSs produced from a Sclerotium-forming fungus (Sclerotium glucanicum DSM 2159) and a yeast (Rhodosporidium babjevae), denoted as scleroglucan (Scl) and EPS-R, respectively. EPS yields of 0.9 ± 0.07 g/L and 1.11 ± 0.4 g/L were obtained from S. glucanicum and R. babjevae, respectively. The physicochemical properties of the EPSs were characterized using infrared spectroscopy and scanning electron microscopy. Further investigations of the biological properties showed that both EPSs were cytocompatible toward the human fibroblast cell line and demonstrated  hemocompatibility. Favorable wound healing capacities of the EPSs (10 mg/mL) were also established via in vivo tests. The present study therefore showed that the EPSs produced by S. glucanicum and R. babjevae have the potential use as biocompatible components for the promotion of dermal wound healing.


Asunto(s)
Ascomicetos , Cicatrización de Heridas , Humanos , Bacterias/metabolismo , Ascomicetos/metabolismo , Suplementos Dietéticos , Línea Celular , Polisacáridos Bacterianos/farmacología , Polisacáridos Bacterianos/metabolismo
13.
Carbohydr Polym ; 284: 119152, 2022 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-35287892

RESUMEN

Fungal exopolysaccharides (EPSs) are natural biopolymers with diverse potential applications in the biomedical, packaging, cosmetic, and food industries. Fungal EPSs are easy to extract and purify polysaccharides that are biodegradable, biocompatible, with low immunogenicity, bioadhesion ability, antibacterial activity, and contain different reactive groups such as hydroxyl, carboxyl, and amine for chemical modifications. Despite fast progress in identifying and characterization fungal EPSs for biomedical applications, i.e., wound healing, drug, and gene delivery, only a few products have been commercialized based on fungal EPSs. This review critically discusses potential biomedical applications of fungi sourced EPSs in tissue engineering (TE), drug and gene delivery.


Asunto(s)
Quitosano , Antibacterianos/química , Antibacterianos/farmacología , Quitosano/química , Hongos , Ingeniería de Tejidos , Cicatrización de Heridas
14.
Front Bioeng Biotechnol ; 9: 748213, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34540818

RESUMEN

Scleroglucan is a non-ionic water-soluble polysaccharide, and has been widely used in the petroleum, food, medicine and cosmetics industries. Currently, scleroglucan is mainly produced by Sclerotium rolfsii. A higher level of scleroglucan (42.0 g/L) was previously obtained with S. rolfsii WSH-G01. However, the production of scleroglucan was reduced despite a higher glucose concentration remaining. Additionally, the molecular weight of scleroglucan was large, thus restricted its application. In this study, by adjusting the state of seeds inoculated, the degradation issue of scleroglucan during the fermentation process was solved. By comparing different fed-batch strategies, 66.6 g/L of scleroglucan was harvested by a two-dose fed-batch mode, with 53.3% glucose conversion ratio. To modify the molecular weight of scleroglucan, a combination method with HCl and high-pressure homogenization treatment was established. Finally, scleroglucan with molecular weight of 4.61 × 105 Da was obtained. The developed approaches provide references for the biosynthesis and molecular weight modification of polysaccharides.

15.
Bioresour Technol ; 335: 125286, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34022479

RESUMEN

Endo-ß-1,3-glucanase with high specific activity is a prerequisite for enzymatic preparation of valuable ß-oligoglucosides. Heterologous expression in Pichia pastoris GS115 with error-prone PCR technology was implemented, and the mutant strain 7 N12 was obtained. The mutant endo-ß-1,3-glucanase showed efficient specific activities for degrading curdlan (366 U mg-1) and scleroglucan (274.5 U mg-1). Thereafter, one-step production of functional branched oligoglucosides was established with coupled fermentation of Pichia pastoris and Sclerotium rolfsii. During the fermentation process, the endo-ß-1,3-glucanase secreted by Pichia pastoris GS115 can efficiently hydrolyse scleroglucan metabolized by Sclerotium rolfsii WSH-G01. The maximum yields of ß-oligoglucosides in the shake flasks and 7-L bioreactor reached 1.73 g L-1 and 12.71 g L-1, respectively, with polymerization degrees of 2-17. The successful implementation of heterologous expression with error-prone PCR and the coupled fermentation simplified the multi-step enzymatic ß-oligoglucoside preparation procedures, which makes it a potential strategy for industrial production of functional oligosaccharides.


Asunto(s)
Pichia , Saccharomycetales , Basidiomycota , Fermentación , Pichia/genética , Proteínas Recombinantes/genética
16.
Carbohydr Polym ; 260: 117505, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712177

RESUMEN

Exopolysaccharide (EPS) secretion by Sclerotium rolfsii ATCC 201126 in submerged cultures, already identified as high-osmolarity responsive, was assessed by reducing C-source without compromising EPS yields. A designed medium with 80 g sucrose L-1 (MOPT80) was tested at 3 L-bioreactor scale at different temperature, agitation, aeration and pH (uncontrolled vs. controlled) values. Optimal operative conditions (200 rpm, 28 °C, 0.5 vvm and initial pH -pHi- 4.5) were validated, as well as the possibility to work at pHi 5.5 to reduce biomass production. Purified EPSs produced in MOPT80 at optimal and other valid operative conditions exhibited refined grade (<1 % proteins and ash, 3-4 % reducing sugars, 87-99 % total sugars). EPS purity, MW and rheological parameters led to discourage pH controlled at 4.5. Relatively constant MW (6-8 × 106 Da) and outstanding viscosifying ability were found. Polyphasic EPS analysis (titre, purity, macromolecular features and rheological fitness) would support to properly select production conditions.


Asunto(s)
Basidiomycota/crecimiento & desarrollo , Reactores Biológicos , Glucanos/metabolismo , Basidiomycota/metabolismo , Biomasa , Reactores Biológicos/economía , Medios de Cultivo/química , Concentración de Iones de Hidrógeno , Reología , Temperatura
17.
Fungal Biol Biotechnol ; 8(1): 1, 2021 Feb 18.
Artículo en Inglés | MEDLINE | ID: mdl-33602329

RESUMEN

BACKGROUND: Sclerotium rolfsii is a potent producer of many secondary metabolites, one of which like scleroglucan is an exopolysaccharide (EPS) appreciated as a multipurpose compound applicable in many industrial fields. RESULTS: Aspartate transaminase (AAT1) catalyzes the interconversion of aspartate and α-ketoglutarate to glutamate and oxaloacetate. We selected AAT1 in the oxalate metabolic pathway as a target of CRISPR/Cas9. Disruption of AAT1 leads to the accumulation of oxalate, rather than its conversion to α-ketoglutarate (AKG). Therefore, AAT1-mutant serves to lower the pH (pH 3-4) so as to increase the production of the pH-sensitive metabolite scleroglucan to 21.03 g L-1 with a productivity of up to 0.25 g L-1·h-1. CONCLUSIONS: We established a platform for gene editing that could rapidly generate and select mutants to provide a new beneficial strain of S. rolfsii as a scleroglucan hyper-producer, which is expected to reduce the cost of controlling the optimum pH condition in the fermentation industry.

18.
Sheng Wu Gong Cheng Xue Bao ; 37(1): 207-217, 2021 Jan 25.
Artículo en Chino | MEDLINE | ID: mdl-33501802

RESUMEN

Scleroglucan is a high-molecular water-soluble microbial exopolysaccharide and mainly applied in the fields of petroleum, food, medicine and cosmetics. The high molecular weight of scleroglucan produced by microbial fermentation leads to low solubility, high viscosity and poor dispersibility, thus bringing a series of difficulties to extraction, preservation and application. It is important to explore suitable degradation method to adjust the molecular weight of scleroglucan for expanding its industrial application. Taking Sclerotium rolfsii WSH-G01 as a model strain, in which functional annotations of the glucanase genes were conducted by whole genome sequencing. Based on design of culture system for culture system for differential expression of ß-glucanase, endogenous ß-glucanase genes in S. rolfsii WSH-G01 were excavated by transcriptomics analysis. Functions of these potential hydrolases were further verified. Finally, 14 potential endogenous hydrolase genes were obtained from S. rolfsii. After heterologous overexpression in Pichia pastoris, 10 soluble enzymes were obtained and 5 of them had the activity of laminarin hydrolysis by SDS-PAGE and enzyme activity analysis. Further investigation of the 5 endogenous hydrolases on scleroglucan degradation showed that enzyme GME9860 has positive hydrolysis effect. The obtained results provide references not only for obtaining low and medium molecular weight of scleroglucan with enzymatic hydrolysis, but also for producing different molecular weight of scleroglucan during S. rolfsii fermentation process with metabolic engineering.


Asunto(s)
Basidiomycota , Basidiomycota/genética , Glucanos , Hidrólisis , Saccharomycetales
19.
Int J Biol Macromol ; 163: 1196-1207, 2020 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-32622769

RESUMEN

Athelia rolfsii TEMG (MH 236106) an exopolysaccharide (EPS) producing fungal strain was isolated and identified. Extraction, purification, characterization, antimicrobial, antioxidant, antiviral and antitumor activities of the polysaccharide were investigated. It characterized as a homopolysaccharide of glucose with a molecular weight of 345.622 kDa. The identification of the polysaccharide was conducted using scanning electron microscopy, energy dispersive X-ray analysis, 1H and 13C NMR spectra. The existence of ß-1,3 and ß-1,6 linkages suggests that EPS could be scleroglucan. The purified scleroglucan showed considerable antibacterial and antioxidant activities. The results indicated that, there was no cytotoxicity on normal cell (W138) and no effect on tumor cell lines including HepG2 and PC3 showing IC50 of 5096.83, 5885.80 and 4803.90 µg/mL, respectively. The results showed also that Sclg could reduce the cytopathic effect by 50% (EC50) at 15 and 50 µg/mL of herpes simplex virus type-1 (HSV-1) and influenza virus (H5N1), respectively.


Asunto(s)
Basidiomycota/química , Glucanos/química , Glucanos/farmacología , Basidiomycota/clasificación , Basidiomycota/genética , Supervivencia Celular/efectos de los fármacos , Fenómenos Químicos , Humanos , Estructura Molecular , Peso Molecular , Monosacáridos/química , Polisacáridos/química , Polisacáridos/farmacología , Reología , Relación Estructura-Actividad
20.
Int J Biol Macromol ; 162: 781-797, 2020 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-32553980

RESUMEN

The development of thermosensitive smart hydrogels with suitable thermosensitivity is of great importance for various biomedical and pharmaceutical applications. In this study, thermosensitive chitosan/carboxymethylcellulose/scleroglucan/montmorillonite (CHT/CMC/SGL/MMT) nanocomposite hydrogels were prepared for biomedical and pharmaceutical usages and characterized by using rheology, FTIR, SEM, EDX, TEM, XRD, TGA and swelling measurements. Exfoliated distribution of MMT in the network structure of hydrogels proved by XRD and TEM analyses caused a decrease in the pore size of hydrogels. Phase transition temperature of thermosensitive hydrogels was determined precisely by rheological measurements. In the presence of 5% MMT within hydrogel matrix, the gelling temperature of Sample 9 exhibited a decrease from 32.0 °C to 25.3 °C. It was found from TGA that among the CHT/CMC/SGL/MMT hydrogel materials the hydrogel system containing 5% MMT showed the highest decomposition temperature (175 °C). Furthermore, all hydrogel materials exhibited non-Fickian swelling behavior in distilled water and basic medium. The addition of MMT into the hydrogel matrix caused a significant decrease in the swelling amount of thermosensitive hydrogels. The results of this study indicate that thermosensitive CHT/CMC/SGL/MMT hydrogel materials may have potential applications in drug delivery, wound dressing and tissue engineering.


Asunto(s)
Bentonita/química , Carboximetilcelulosa de Sodio/química , Quitosano/química , Glucanos/química , Hidrogeles/química , Nanogeles/química , Porosidad , Reología , Temperatura
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