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Frontiers in Cellular and Infection... 2023Mucoromycosis is a highly aggressive angio-invasive disease of humans caused by fungi in the zygomycete order, Mucorales. While is the principal agent of mucoromycosis,...
Mucoromycosis is a highly aggressive angio-invasive disease of humans caused by fungi in the zygomycete order, Mucorales. While is the principal agent of mucoromycosis, other Mucorales fungi including , , , , and are able to cause life-threatening rhino-orbital-cerebral, pulmonary, gastro-intestinal and necrotising cutaneous infections in humans. Diagnosis of the disease currently relies on non-specific CT, lengthy and insensitive culture from invasive biopsy, and time-consuming histopathology of tissue samples. At present, there are no rapid antigen tests that detect Mucorales-specific biomarkers of infection, and which allow point-of-care diagnosis of mucoromycosis. Here, we report the development of an IgG2b monoclonal antibody (mAb), TG11, which binds to extracellular polysaccharide (EPS) antigens of between 20 kDa and 250 kDa secreted during hyphal growth of Mucorales fungi. The mAb is Mucorales-specific and does not cross-react with other yeasts and molds of clinical importance including , , , , and species. Using the mAb, we have developed a Competitive lateral-flow device that allows rapid (30 min) detection of the EPS biomarker in human serum and bronchoalveolar lavage (BAL), with a limit of detection (LOD) in human serum of ~100 ng/mL serum (~224.7 pmol/L serum). The LFD therefore provides a potential novel opportunity for detection of mucoromycosis caused by different Mucorales species.
Topics: Humans; Mucorales; Antibodies, Monoclonal; Aspergillus; Fusarium; Biomarkers
PubMed: 38145040
DOI: 10.3389/fcimb.2023.1305662 -
Journal of Biochemistry May 2001Site-directed mutagenesis was carried out to investigate the functional roles of amino acid residues of Rhizomucor pusillus pepsin (RMPP) in substrate-binding and...
Replacements of amino acid residues at subsites and their effects on the catalytic properties of Rhizomucor pusillus pepsin, an aspartic proteinase from Rhizomucor pusillus.
Site-directed mutagenesis was carried out to investigate the functional roles of amino acid residues of Rhizomucor pusillus pepsin (RMPP) in substrate-binding and catalysis. Mutations of two amino acid residues, E13 in the S3 subsite and N219 in the S3/S4 subsites, caused marked changes in kinetic parameters for two substrate peptides with different sequences. Further site-directed mutagenesis at E13 suggested that E13 plays a critical role in forming the correct hydrogen bond network around the active center. In the crystal structure of Rhizomucor miehei pepsin (RMMP), which is an aspartic proteinase produced by Rhizomucor miehei and shows 81% amino acid identity to RMPP, the Oepsilon atom of N219 forms a hydrogen bond with the N-H of isovaline in pepstatin A, a statine-type inhibitor, at the P3 position, suggesting that the loss of the hydrogen bond causes an unfavorable arrangement of the P3 residue. Among the mutants constructed, the E13A mutant showed a 5-fold increase in the ratio of clotting versus proteolytic activity without significant loss of clotting activity. This mutant may present a promising candidate for a useful milk coagulant.
Topics: Amino Acid Substitution; Aspartic Acid Endopeptidases; Catalysis; Fungal Proteins; Hydrogen Bonding; Kinetics; Mutagenesis, Site-Directed; Pepsin A; Rhizomucor
PubMed: 11328603
DOI: 10.1093/oxfordjournals.jbchem.a002921 -
Applied and Environmental Microbiology Apr 2020Chinese (fermentation starter) provides saccharifying enzymes for baijiu (Chinese liquor) fermentation, which undergoes a simultaneous saccharification and fermentation...
Chinese (fermentation starter) provides saccharifying enzymes for baijiu (Chinese liquor) fermentation, which undergoes a simultaneous saccharification and fermentation process. However, the key saccharifying enzymes associated with alcoholic fermentation from and their effects on ethanol production remain poorly understood. In this study, we identified 51 carbohydrate hydrolases in baijiu fermentation by metaproteomics analysis. Through source-tracking analysis, approximately 80% of carbohydrate hydrolases in the baijiu fermentation were provided by Among these enzymes, alpha-amylase (EC 3.2.1.1) and glucoamylase (EC 3.2.1.3), from , , and , were positively related to starch hydrolysis and ethanol production, indicating that they were the key saccharifying enzymes associated with alcoholic fermentation in the baijiu fermentation. Moreover, a combined mixture of alpha-amylase and glucoamylase (in a ratio of 1:6, wt/wt) enhanced ethanol production in a simulative baijiu fermentation under laboratory conditions. This result revealed a synergistic effect of multiple saccharifying enzymes on ethanol production in baijiu fermentation. Our study provides a potential approach to improve the efficiency of saccharification and alcoholic fermentation by optimizing the profile of saccharifying enzymes for fermentation of baijiu and other beverages. starter provides enzymes to the simultaneous saccharification and fermentation process of baijiu (Chinese liquor) production; however, the key saccharifying enzymes associated with alcoholic fermentation from and their effects on ethanol production remain unclear. We confirmed that was the main source of carbohydrate hydrolases for baijiu fermentation and identified two types of saccharifying enzymes from multiple microbes as the key enzymes associated with alcoholic fermentation. Moreover, a proper combination of multiple saccharifying enzymes could enhance ethanol production in baijiu fermentation. This combination provides an approach to optimize the profile of saccharifying enzymes for enhancing ethanol production in baijiu and other food fermentations.
Topics: Alcoholic Beverages; Animals; Bacteria; Carbohydrate Metabolism; China; Fermentation; Fungi; Hydrolases; Microbiota; Mycobiome
PubMed: 32060021
DOI: 10.1128/AEM.00013-20 -
In Vitro Activity of Isavuconazole and Comparators against Clinical Isolates of the Mucorales Order.Antimicrobial Agents and Chemotherapy Dec 2015The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of...
The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of amphotericin B, posaconazole, and voriconazole. Seventy-two isolates were included: 12 of Lichtheimia corymbifera, 5 of Lichtheimia ramosa, 5 of group I and 9 of group II of Mucor circinelloides, 9 of Rhizomucor pusillus, 26 of Rhizopus microsporus, and 6 of Rhizopus oryzae. Species identification was confirmed by internal transcribed spacer (ITS) sequencing. EUCAST MICs were read on day 1 (EUCAST-d1) and day 2 (EUCAST-d2), and CLSI MICs were read on day 2 (CLSI-d2). Isavuconazole MIC50s (range) (mg/liter) by EUCAST-d1, CLSI-d2, and EUCAST-d2 were 1 (0.125 to 16), 1 (0.125 to 2), and 4 (0.5 to >16), respectively, across all isolates. The similar values for comparator drugs were as follows: posaconazole, 0.25 (≤ 0.03 to >16), 0.25 (0.06 to >16), and 1 (0.06 to >16); amphotericin, 0.06 (≤ 0.03 to 0.5), 0.06 (≤ 0.03 to 0.25), and 0.125 (≤ 0.03 to 1); voriconazole, 16 (2 to >16), 8 (1 to >16), and >16 (8 to >16), respectively. Isavuconazole activity varied by species: Lichtheimia corymbifera, 1 (0.5 to 2), 1 (1 to 2), and 2 (1 to 4); Lichtheimia ramosa, 0.25 (0.125 to 0.5), 1 (0.5 to 2), and 2 (0.5 to 4); Rhizomucor pusillus, 0.5 (0.5 to 1), 1 (0.125 to 1), and 2 (1 to 2); Rhizopus microsporus, 1 (0.5 to 4), 0.5 (0.125 to 1), and 4 (1 to 8); and Rhizopus oryzae, 1 (0.5 to 4), 1 (0.125 to 2), and 4 (0.5 to 8), respectively, were more susceptible than Mucor circinelloides: group I, 8 (4 to 8), 4 (2 to 4), and 16 (2 to 16), respectively, and group II, 8 (1 to 16), 8 (1 to 8), and 16 (4 to >16), respectively. This was also observed for posaconazole. The essential agreement was best between EUCAST-d1 and CLSI-d2 (75% to 83%). Isavuconazole displayed in vitro activity against Mucorales isolates with the exception of Mucor circinelloides. The MICs were in general 1 to 3 steps higher than those for posaconazole. However, in the clinical setting this may be compensated for by the higher exposure at standard dosing.
Topics: Antifungal Agents; Fungi; Humans; Microbial Sensitivity Tests; Mucorales; Mucormycosis; Nitriles; Pyridines; Species Specificity; Triazoles
PubMed: 26438494
DOI: 10.1128/AAC.01919-15 -
Medicina Oral, Patologia Oral Y Cirugia... Dec 2008Mucormycosis is a rare oportunistic infection typically described in diabetic patients with a ketoacidotic status, as well as neutropenic patients. The infection is... (Review)
Review
Mucormycosis is a rare oportunistic infection typically described in diabetic patients with a ketoacidotic status, as well as neutropenic patients. The infection is caused by a group of saprophytic fungi of the class Phycomicetes, being the most frequent ones the Rhizomucor, Rhizopus and Mucor. Its hystological findings include vascular trombosis and tissue necrosis, predominantly in the rino-orbito-cerebral area. Even though the frequency of presentation is very low, given its rapid evolution and severe consequences which include a high mortality rate, it is very important to be aware of the main features of the disease and treat it promptly. Although the diagnosis is based on the high clinical suspect, the computed tomography (CT) and the magnetic resonance image (MRI) plays an important role in determining the extension. The patients should receive treatment in a reference hospital so that a multidisciplinary approach is ensured. In this sense, we present a case of rhino-orbito-cerebralmucormycosis in a diabetic patient, recently treated in our Department. A comprehensive review of the literature has been performed to update the physiopathology and diagnosis. Finally, we describe the different treatment options focusing in the surgical approach, as well as the medical treatment with amphotericine and posaconzole.
Topics: Aged; Brain Diseases; Diabetes Complications; Humans; Male; Mucormycosis; Nose Diseases; Orbital Diseases
PubMed: 19047969
DOI: No ID Found -
Journal of Fungi (Basel, Switzerland) Mar 2021β-Galactosidases of Mucoromycota are rarely studied, although this group of filamentous fungi is an excellent source of many industrial enzymes. In this study, 99...
β-Galactosidases of Mucoromycota are rarely studied, although this group of filamentous fungi is an excellent source of many industrial enzymes. In this study, 99 isolates from the genera , , , , and , were screened for their β-galactosidase activity using a chromogenic agar approach. Ten isolates from the best producers were selected, and the activity was further investigated in submerged (SmF) and solid-state (SSF) fermentation systems containing lactose and/or wheat bran substrates as enzyme production inducers. Wheat bran proved to be efficient for the enzyme production under both SmF and SSF conditions, giving maximum specific activity yields from 32 to 12,064 U/mg protein and from 783 to 22,720 U/mg protein, respectively. Oligosaccharide synthesis tests revealed the suitability of crude β-galactosidases from Szeged Microbiological Collection (SZMC) 11360 and SZMC 11025 to catalyze transgalactosylation reactions. In addition, the crude enzyme extracts had transfructosylation activity, resulting in the formation of fructo-oligosaccharide molecules in a sucrose-containing environment. The maximal oligosaccharide concentration varied between 0.0158 and 2.236 g/L depending on the crude enzyme and the initial material. Some oligosaccharide-enriched mixtures supported the growth of probiotics, indicating the potential of the studied enzyme extracts in future prebiotic synthesis processes.
PubMed: 33808917
DOI: 10.3390/jof7030229 -
Journal of Oleo Science 2013A mixture of lard and rapeseed oil (1:1, wt/wt) was interesterified using immobilized lipases from Rhizomucor miehei (Lipozyme RM IM) and Candida antarctica (Novozym...
A mixture of lard and rapeseed oil (1:1, wt/wt) was interesterified using immobilized lipases from Rhizomucor miehei (Lipozyme RM IM) and Candida antarctica (Novozym 435) as catalysts. Enzymatic interesterifications were carried out at 60°C for 8 h with Lipozyme RM IM or at 80°C for 4 h with Novozym 435. The biocatalyst doses were kept constant (8 wt-%). The starting blend was quantitatively separated by column chromatography into pure triacylglycerol fraction (98.5%), and a nontriacylglycerol fraction containing free fatty acids (0.3%) and of mono- and diacylglycerols (1.2%). It was found that after interesterification the contents of free fatty acids and of mono- and diacylglycerols increased to 3.5% and 6.3% or to 1.5% and 4.5% when Lipozyme RM IM and Novozym 435 were used, respectively.The slip melting temperatures and solid fat contents of the triacylglycerol fractions separated from interesterified samples were lower compared with the nonesterified blend. The sn-2 and sn-1,3 distribution of fatty acids in the triacylglycerol fractions before and after interesterification were determined. The compositions of fatty acids at sn-2 were near statistical (33.3%) when Novozym 435 was used. When Lipozyme RM IM was used, the fatty acid compositions at the sn-2 position remained practically unchanged, compared with the starting blend. The changes in molecular structures of fat components due to interesterification have greatly influenced on the melting profiles of products as illustrated by the DSC melting scans. The interesterified fats and isolated triacylglycerols had reduced oxidative stabilities, as assessed by Dynamic DSC and Isothermal PDSC measurements. The Arrhenius kinetic parameters for fats oxidation based on DSC and PDSC measurements were calculated.
Topics: Calorimetry, Differential Scanning; Candida; Dietary Fats; Diglycerides; Esterification; Fats; Fatty Acids; Fatty Acids, Monounsaturated; Lipase; Oxidation-Reduction; Plant Oils; Rapeseed Oil; Regression Analysis; Rhizomucor; Temperature; Triglycerides
PubMed: 23535304
DOI: 10.5650/jos.62.187 -
Frontiers in Cellular and Infection... 2024Mucormycosis is an uncommon invasive fungal infection that has a high mortality rate in patients with severe underlying diseases, which leads to immunosuppression. Due...
BACKGROUND
Mucormycosis is an uncommon invasive fungal infection that has a high mortality rate in patients with severe underlying diseases, which leads to immunosuppression. Due to its rarity, determining the incidence and optimal treatment methods for mucormycosis in children is challenging. Metagenomic next-generation sequencing (mNGS) is a rapid, precise and sensitive method for pathogen detection, which helps in the early diagnosis and intervention of mucormycosis in children. In order to increase pediatricians' understanding of this disease, we conducted a study on the clinical features of mucormycosis in children and assessed the role of mNGS in its diagnosis.
METHODS
We retrospectively summarized the clinical data of 14 children with mucormycosis treated at the First Affiliated Hospital of Zhengzhou University from January 2020 to September 2023.
RESULTS
Of the 14 cases, 11 case of mucormycosis were classified as probable, and 3 cases were proven as mucormycosis. Most children (85.71%) had high-risk factors for mucormycosis. All 14 children had lung involvement, with 5 cases of extrapulmonary dissemination. Among the 14 cases, 4 cases underwent histopathological examination of mediastinum, lung tissue or kidney tissue, in which fungal pathogens were identified in 3 patients. Fungal hyphae was identified in 3 cases of mucormycosis, but only 1 case yielded a positive culture result. All patients underwent mNGS testing with samples from blood (8/14), bronchoalveolar lavage fluid (6/14), and tissue (1/14). mNGS detected fungi in all cases: 7 cases had , 4 cases had , 3 cases had , 1 case had , and 1 case had . Coinfections were found with in 3 cases, bacteria in 3 cases, and viruses in 5 cases.
CONCLUSION
Children with mucormycosis commonly exhibit non-specific symptoms like fever and cough during the initial stages. Early diagnosis based on clinical symptoms and imaging is crucial in children suspected of having mucormycosis. mNGS, as a supplementary diagnostic method, offers greater sensitivity and shorter detection time compared to traditional mucormycosis culture or histopathological testing. Additionally, mNGS enables simultaneous detection of bacteria and viruses, facilitating timely and appropriate administration of antibiotics and thereby enhancing patient outcomes.
Topics: Humans; Mucormycosis; High-Throughput Nucleotide Sequencing; Male; Female; Child; Child, Preschool; Metagenomics; Retrospective Studies; Infant; Adolescent; Invasive Fungal Infections; China
PubMed: 38915923
DOI: 10.3389/fcimb.2024.1368165 -
Journal of Oleo Science 2021In this study, Candida antarctica lipase B (CALB), Rhizomucor miehei lipase (RML) and Lecitase Ultra (LU) were immobilized onto the mesoporous silica SBA-15. The...
In this study, Candida antarctica lipase B (CALB), Rhizomucor miehei lipase (RML) and Lecitase Ultra (LU) were immobilized onto the mesoporous silica SBA-15. The glycerolysis performance of the obtained supported lipases (lipase@SBA-15) in solvent systems was carefully investigated. LU@SBA-15 exhibited good glycerolysis performance in solvent-free system, with diacylglycerols (DAG) content and triacylglycerols (TAG) conversion at 52.4 and 98.6% respectively obtained after 12 h reaction at 60°C. CALB@SBA-15 showed good glycerolysis activity in tert-pentanol and tert-butanol systems, with TAG conversion over 90% obtained. In addition, the present CALB@SBA-15 exhibited selectivity for monoacylglycerols (MAG) production, with glycerol to TAG molar ratio increased to 3:1, MAG content over 80% and TAG conversion over 99% could be obtained from both tert-pentanol and tert-butanol systems. However, RML@SBA-15 showed low glycerolysis activity neither in solvent nor in solvent-free systems. The present results favor the practical enzymatic design for MAG and DAG production.
Topics: Butanols; Diglycerides; Enzymes, Immobilized; Fungal Proteins; Glycerol; Lipase; Monoglycerides; Pentanols; Rhizomucor; Silicon Dioxide; Solvents; Time Factors; Triglycerides
PubMed: 33658468
DOI: 10.5650/jos.ess20228 -
Food Chemistry: X Jun 2023Traditional produced around Winter Solstice has higher quality and a more harmonious aroma. To investigate the variations of volatile metabolites and microbial...
Dynamic analysis of volatile metabolites and microbial community and their correlations during the fermentation process of traditional (Chinese rice wine) produced around Winter Solstice.
Traditional produced around Winter Solstice has higher quality and a more harmonious aroma. To investigate the variations of volatile metabolites and microbial communities during fermentation, gas chromatography-ion migration chromatography (GC-IMS), gas chromatographymass spectroscopy (GC-MS) and highthroughput sequencing were employed. Aroma compounds results showed that alcohols and phenols increased before 45 days of fermentation and then decreased after 45 days, while esters gradually increased. Fungal genera , , and were dominant, whereas , and were the dominant bacterial genera in the late stage. In addition, 11 genera such as , and (|r| > 0.6, < 0.05) may contributed to traditional ecosystem stability. Moreover, correlation analysis indicated the dominant microorganisms (, , , and ) were positively correlated with key compounds. These results provided theoretical guidance for further study on the flavor regulation of traditional via microbial community level and microbial augmentation.
PubMed: 36993869
DOI: 10.1016/j.fochx.2023.100620