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International Journal of Molecular... Mar 2023Truffles are known worldwide for their peculiar taste, aroma, and nutritious properties, which increase their economic value. However, due to the challenges associated...
Truffles are known worldwide for their peculiar taste, aroma, and nutritious properties, which increase their economic value. However, due to the challenges associated with the natural cultivation of truffles, including cost and time, submerged fermentation has turned out to be a potential alternative. Therefore, in the current study, the cultivation of in submerged fermentation was executed to enhance the production of mycelial biomass, exopolysaccharides (EPSs), and intracellular polysaccharides (IPSs). The mycelial growth and EPS and IPS production was greatly impacted by the choice and concentration of the screened carbon and nitrogen sources. The results showed that sucrose (80 g/L) and yeast extract (20 g/L) yielded maximum mycelial biomass (5.38 ± 0.01 g/L), EPS (0.70 ± 0.02 g/L), and IPS (1.76 ± 0.01 g/L). The time course analysis of truffle growth revealed that the highest growth and EPS and IPS production was observed on the 28th day of the submerged fermentation. Molecular weight analysis performed by the gel permeation chromatography method revealed a high proportion of high-molecular-weight EPS when 20 g/L yeast extract was used as media and the NaOH extraction step was carried out. Moreover, structural analysis of the EPS using Fourier-transform infrared spectroscopy (FTIR) confirmed that the EPS was β-(1-3)-glucan, which is known for its biomedical properties, including anti-cancer and anti-microbial activities. To the best of our knowledge, this study represents the first FTIR analysis for the structural characterization of β-(1-3)-glucan (EPS) produced from grown in submerged fermentation.
Topics: Fermentation; Molecular Weight; Polysaccharides; Glucans
PubMed: 36902305
DOI: 10.3390/ijms24054875 -
Journal of Dermatological Science Jul 2022The biological functions of Hyaluronic acid are related to its molecular weight and binding to its receptor, Toll-like receptor4 (TLR4) or CD44. Recent studies have...
BACKGROUND
The biological functions of Hyaluronic acid are related to its molecular weight and binding to its receptor, Toll-like receptor4 (TLR4) or CD44. Recent studies have shown that low-molecular-weight Hyaluronic acid (LMW-HA) exhibits proinflammatory effects, while high-molecular-weight Hyaluronic acid (HMW-HA) functions as an anti-inflammatory factor. UVB-induced epidermal inflammation is mainly mediated by endogenous molecules, such as damage-associated molecular patterns (DAMPs), that cause severe skin damage by activating TLR signaling pathways.
OBJECTIVE
Since both LMW- and HMW-HA have inhibitory functions on TLR-mediated macrophage inflammation, HA is assumed to suppress UVB-induced DAMP-mediated inflammation in the skin. In this study, both Ultra- low-molecular-weight Hyaluronic acid (uLMW-HA) and HMW-HA were found to inhibit UVB-induced keratinocyte inflammation.
METHODS
HaCaT cells were treated with medium containing Hyaluronic acid at the appropriate concentration after 15 mJ/cm irradiation. Secreted protein levels were determined with ELISA kits. Expression levels of proteins downstream of TLR4 were detected by Simple Western system.
RESULTS
By competitively binding to TLR4, uLMW-HA downregulated Calprotectin-induced TRAF6 expression, which might be the direct process by which uLMW-HA decreased UVB-induced IL-6 secretion. Reduced CD44 variant (CD44v) expression in keratinocytes attenuated the inhibitory effect of both uLMW-HA and HMW-HA on UVB-induced inflammation, which indicated the involvement of CD44v in HA-regulated anti-inflammatory activity.
CONCLUSION
Overall, this research indicates that Hyaluronic acid is more than a moisturizer; it is also a biologically effective material that can prevent the excessive skin inflammation caused in daily life, especially in the late stages after sunburn.
Topics: Anti-Inflammatory Agents; Humans; Hyaluronic Acid; Inflammation; Keratinocytes; Leukocyte L1 Antigen Complex; Molecular Weight; Toll-Like Receptor 4
PubMed: 35717315
DOI: 10.1016/j.jdermsci.2022.06.001 -
European Journal of Pharmaceutical... Jan 2022Hyaluronic acid (HA) is widely adopted to fabricate dissolving microneedles for transdermal drug delivery applications, yet the structure-activity relationship between...
Hyaluronic acid (HA) is widely adopted to fabricate dissolving microneedles for transdermal drug delivery applications, yet the structure-activity relationship between molecular weight of HA and transdermal delivery efficiency of microneedles (HA-MNs) has not been fully explored, particularly in the transdermal delivery of small molecule drugs. Herein, we report the fabrication of three types of HA-MNs of various molecular weights (10k, 74k and 290k Da), which incorporate rhodamine B as the model drug. We assess the influence of molecular weight of HA on the mechanical properties of HA-MNs and transdermal delivery of rhodamine B in vitro and in vivo. The mechanical strength of all types of HA-MNs exceeds the minimal force requirement for skin penetration, with the highest values of compression force found in 10k-HA-MN. Interestingly, 74k-HA-MN that owns a medium mechanical strength, exhibits the highest efficiency in transdermal delivery of rhodamine B in a porcine skin and a Franz cell transdermal model. Further in vivo fluorescence imaging of HA-MN-treated mice reveals a tunable transdermal delivery of rhodamine B, which is controllable according to the molecular weight of HA. Importantly, 74k-HA-MN treatment demonstrates the highest initial delivering amount and longest retention time of rhodamine B in mice. In addition, histological examinations of puncture sites of the skin tissues confirm the complete recovery of skin and excellent biocompatibility of HA-MNs.
Topics: Administration, Cutaneous; Animals; Drug Delivery Systems; Hyaluronic Acid; Mice; Molecular Weight; Needles; Skin; Swine
PubMed: 34813921
DOI: 10.1016/j.ejps.2021.106075 -
Iranian Biomedical Journal Nov 2022Background: Hyaluronic acid (HA), a natural polymer with wide applications in biomedicine and cosmetics, is mainly produced by Streptococcal fermentation at industrial...
BACKGROUND
Background: Hyaluronic acid (HA), a natural polymer with wide applications in biomedicine and cosmetics, is mainly produced by Streptococcal fermentation at industrial scale. In the present study, chemical random mutagenesis was used for development of Streptococcus equisimilis group G mutant strains with high HA productivity.
METHODS
Methods: The optimum of the pH of culture condition and cultivation time for HA production by wild strain group G were assessed. At first, two rounds of mutation at different concentrations of NTG was used for mutagenesis. Then, the nonhemolytic and hyaluronidase-negative mutants were screened on the blood and HA agar. HA productivity and molecular weight were determined by carbazole assay, agarose gel electrophoresis and specific staining. Moreover, stability of the high producer mutants was evaluated within 10 generations.
RESULTS
Results: The results showed that the wild-type strain produced 1241 ± 2.1 µg/ml of HA at pH 5.5 and 4 hours of cultivation, while the screened mutants showed a 16.1-45.5% increase in HA production. Two mutant strains, named Gm2-120-21-3 (2470 ± 8.1 µg/ml) and Gm2-120-21-4 (2856 ± 4.2 µg/ml), indicated the highest titer and a consistent production. The molecular weight (Mw) of HA for the mutants was less than 160 kDa, considering as a low Mw HA.
CONCLUSION
Conclusion: The mutant strains producing a low polydisperse, as well as low Mw of HA with high titer might be regarded as potential industrial strains for HA production after further safety investigations.
Topics: Hyaluronic Acid; Molecular Weight; Agar; Streptococcus
PubMed: 36437793
DOI: 10.52547/ibj.3789 -
International Journal of Molecular... Mar 2022Imines or Schiff bases (SB) are formed by the condensation of an aldehyde or a ketone with a primary amine, with the removal of a water molecule. Schiff bases are...
Imines or Schiff bases (SB) are formed by the condensation of an aldehyde or a ketone with a primary amine, with the removal of a water molecule. Schiff bases are central molecules in several biological processes for their ability to form and cleave by small variation of the medium. We report here the controlled hydrolysis of four SBs that may be applied in the fragrance industry, as they are profragrances all containing odorant molecules: methyl anthranilate as primary amine, and four aldehydes (cyclamal, helional, hydroxycitronellal and triplal) that are very volatile odorants. The SB stability was assessed over time by HPLC-MS in neutral or acidic conditions, both in solution and when trapped in low molecular weight gels. Our results demonstrate that it is possible to control the hydrolysis of the Schiff bases in the gel environment, thus tuning the quantity of aldehyde released and the persistency of the fragrance.
Topics: Aldehydes; Amines; Gels; Hydrolysis; Molecular Weight; Odorants; Perfume; Schiff Bases
PubMed: 35328526
DOI: 10.3390/ijms23063105 -
Toxins Jan 2022Here, we report the current status of the bioactive peptides isolated and characterized from mushrooms during the last 20 years, considering 'peptide' a succession from... (Review)
Review
Here, we report the current status of the bioactive peptides isolated and characterized from mushrooms during the last 20 years, considering 'peptide' a succession from to 2 to 100 amino acid residues. According to this accepted biochemical definition, we adopt ~10 kDa as the upper limit of molecular weight for a peptide. In light of this, a careful revision of data reported in the literature was carried out. The search revealed that in the works describing the characterization of bioactive peptides from mushrooms, not all the peptides have been correctly classified according to their molecular weight, considering that some fungal proteins (>10 kDa MW) have been improperly classified as 'peptides'. Moreover, the biological action of each of these peptides, the principles of their isolation as well as the source/mushroom species were summarized. Finally, this review highlighted that these peptides possess antihypertensive, antifungal, antibiotic and antimicrobial, anticancer, antiviral, antioxidant and ACE inhibitory properties.
Topics: Agaricales; Fungal Proteins; Molecular Weight
PubMed: 35202112
DOI: 10.3390/toxins14020084 -
Drug Discovery Today May 2019We present a novel crystallographic screening methodology (MiniFrags) that employs high-concentration aqueous soaks with a chemically diverse and... (Review)
Review
We present a novel crystallographic screening methodology (MiniFrags) that employs high-concentration aqueous soaks with a chemically diverse and ultra-low-molecular-weight library (heavy atom count 5-7) to identify ligand-binding hot and warm spots on proteins. We propose that MiniFrag screening represents a highly effective method for guiding optimisation of fragment-derived lead compounds or chemical tools and that the high screening hit rates reflect enhanced sampling of chemical space.
Topics: Crystallography; Drug Design; Ligands; Molecular Weight; Small Molecule Libraries
PubMed: 30878562
DOI: 10.1016/j.drudis.2019.03.009 -
Analytical and Bioanalytical Chemistry Mar 2011Size, weight and position are three of the most important parameters that describe a molecule in a biological system. Ion mobility spectrometry is capable of separating... (Review)
Review
Size, weight and position are three of the most important parameters that describe a molecule in a biological system. Ion mobility spectrometry is capable of separating molecules on the basis of their size or shape, whereas imaging mass spectrometry is an effective tool to measure the molecular weight and spatial distribution of molecules. Recent developments in both fields enabled the combination of the two technologies. As a result, ion-mobility-based imaging mass spectrometry is gaining more and more popularity as a (bio-)analytical tool enabling the determination of the size, weight and position of several molecules simultaneously on biological surfaces. This paper reviews the evolution of ion-mobility-based imaging mass spectrometry and provides examples of its application in analytical studies of biological surfaces.
Topics: Animals; Humans; Ions; Mass Spectrometry; Molecular Weight
PubMed: 21225246
DOI: 10.1007/s00216-010-4644-1 -
Journal of Chromatography. B,... Nov 2022High molecular weight (HMW) species are product-related variants that may impact therapeutic product safety and efficacy. Therefore, HMW species and aggregates are...
High molecular weight (HMW) species are product-related variants that may impact therapeutic product safety and efficacy. Therefore, HMW species and aggregates are considered critical quality attributes and their levels should be closely monitored and controlled during drug development, commercial manufacturing, and shelf-life storage period for therapeutic monoclonal antibody drug products. Various biophysical and analytical methods have been developed to characterize the HMW species to understand their mechanisms of formation and assess potential product risk. However, host cell protein (HCP) analysis has seldom been conducted to characterize the impurities in aggregates. In this work, HCP analysis of enriched HMW species and drug substance (DS) from five different monoclonal antibodies (mAbs) was performed. More HCPs are identified in the enriched HMW than in the DS, thus demonstrating a potential interaction between HCPs and HMW. Certain HCPs, including commonly detected HCPs and problematic HCPs, were enriched in HMW fractions. Especially, the most abundant HCP from mAb1, CC motif chemokine, was 46 times more abundant in enriched HMW than DS. The enriched HMW was further fractionated into enriched dimers and enriched very HMW (vHMW) fractions. The CC motif chemokine was found to interact mainly with mAb1 dimer species rather than vHMW fraction. Removing the HMW species from mAb1 significantly decreased the CC motif chemokine level in the final mAb1 DS. Our findings demonstrate that some HCPs are more preferentially bound to HMW species and this finding may provide a new opportunity for removing HCPs in downstream purification steps.
Topics: Animals; Antibodies, Monoclonal; CHO Cells; Chemokines; Cricetinae; Cricetulus; Molecular Weight
PubMed: 36115198
DOI: 10.1016/j.jchromb.2022.123448 -
International Journal of Biological... Jun 2023Cationic polyelectrolytes (PEs) are commonly used additives in manufacturing of cellulose based products such as regenerated fibers and paper to tailor their product...
Cationic polyelectrolytes (PEs) are commonly used additives in manufacturing of cellulose based products such as regenerated fibers and paper to tailor their product properties. Here we are studying the adsorption of poly(diallyldimethylammonium chloride), PD, on cellulose, using in situ surface plasmon resonance spectroscopy (SPR) measurements. We employ model surfaces from regenerated cellulose xanthate (CX) and trimethylsilyl cellulose (TMSC), mimicking industrially relevant regenerated cellulose substrates. The effects of the PDs molecular weight were strongly depending on the ionic strength and type of electrolyte (NaCl vs CaCl). Without electrolytes, the adsorption was monolayer-type, i.e. independent of molecular weight. At moderate ionic strength, adsorption increased due to more pronounced PE coiling, while at high ionic strength electrostatic shielding strongly reduced adsorption of PDs. Results exhibited pronounced differences for the chosen substrates (cellulose regenerated from xanthate (CX) vs. regenerated from trimethylsilyl cellulose, TMSC). Consistently higher adsorbed amounts of the PD were determined on CX surfaces compared TMSC. This can be attributed to a more negative zeta potential, a higher AFM roughness and a higher degree of swelling (investigated by QCM-D) of the CX substrates.
Topics: Adsorption; Molecular Weight; Surface Properties; Cellulose; Electrolytes
PubMed: 37011749
DOI: 10.1016/j.ijbiomac.2023.124286