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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 -
Molecules (Basel, Switzerland) Mar 2023Crude olive pomace oil (OPO) is a by-product of olive oil extraction. In this study, low-calorie structured triacylglycerols (TAGs) were produced by acidolysis of crude...
Crude olive pomace oil (OPO) is a by-product of olive oil extraction. In this study, low-calorie structured triacylglycerols (TAGs) were produced by acidolysis of crude OPO with medium-chain fatty acids (caprylic, C8:0; capric, C10:0) or interesterification with their ethyl ester forms (C8EE, C10EE). These new TAGs present long-chain fatty acids (L) at position -2 and medium-chain fatty acids (M) at positions -1,3 (MLM). Crude OPO exhibited a high acidity (12.05-28.75% free fatty acids), and high contents of chlorophylls and oxidation products. Reactions were carried out continuously in a packed-bed bioreactor for 70 h, using -1,3 regioselective commercial immobilized lipases ( lipase, Lipozyme TL IM; and lipase, Lipozyme RM IM), in solvent-free media at 40 °C. Lipozyme RM IM presented a higher affinity for C10:0 and C10EE. Lipozyme TL IM preferred C10:0 over C8:0 but C8EE over C10EE. Both biocatalysts showed a high activity and operational stability and were not affected by OPO acidity. The New TAG yields ranged 30-60 and the specific productivity ranged 0.96-1.87 g NewTAG/h.g biocatalyst. Lipozyme RM IM cost is more than seven-fold the Lipozyme TL IM cost. Therefore, using Lipozyme TL IM and crude acidic OPO in a continuous bioreactor will contribute to process sustainability for structured lipid production by lowering the cost of the biocatalyst and avoiding oil refining.
Topics: Olea; Plant Oils; Dietetics; Dietary Fats; Triglycerides; Fatty Acids; Olive Oil; Lipase; Esterification; Enzymes, Immobilized
PubMed: 36985609
DOI: 10.3390/molecules28062637 -
Applied and Environmental Microbiology Mar 2023Cavities are created by hydrophobic interactions between residue side chain atoms during the folding of enzymes. Redesigning cavities can improve the thermostability and...
Cavities are created by hydrophobic interactions between residue side chain atoms during the folding of enzymes. Redesigning cavities can improve the thermostability and catalytic activity of the enzyme; however, the synergistic effect of cavities remains unclear. In this study, Rhizomucor miehei lipase (RML) was used as a model to explore volume fluctuation and spatial distribution changes of the internal cavities, which could reveal the roles of internal cavities in the thermostability and catalytic activity. We present an inside out cavity engineering (CE) strategy based on computational techniques to explore how changes in the volumes and spatial distribution of cavities affect the thermostability and catalytic activity of the enzyme. We obtained 12 single-point mutants, among which the melting temperatures () of 8 mutants showed an increase of more than 2°C. Sixteen multipoint mutations were further designed by spatial distribution rearrangement of internal cavities. The of the most stable triple variant, with mutations including T21V (a change of T to V at position 21), S27A, and T198L (T21V/S27A/T198L), was elevated by 11.0°C, together with a 28.7-fold increase in the half-life at 65°C and a specific activity increase of 9.9-fold (up to 5,828 U mg), one of the highest lipase activities reported. The possible mechanism of decreased volumes and spatial rearrangement of the internal cavities improved the stability of the enzyme, optimizing the outer substrate tunnel to improve the catalytic efficiency. Overall, the inside out computational redesign of cavities method could help to deeply understand the effect of cavities on enzymatic stability and activity, which would be beneficial for protein engineering efforts to optimize natural enzymes. In the present study, R. miehei lipase, which is widely used in various industries, provides an opportunity to explore the effects of internal cavities on the thermostability and catalytic activity of enzymes. Here, we execute high hydrostatic pressure molecular dynamics (HP-MD) simulations to screen the critical internal cavity and reshape the internal cavities through site-directed mutation. We show that as the global internal cavity volume decreases, cavity rearrangement can improve the stability of the protein while optimizing the substrate channel to improve the catalytic efficiency. Our results provide significant insights into understanding the mechanism of action of the internal cavity. Our strategy is expected to be applied to other enzymes to promote increases in thermostability and catalytic activity.
Topics: Lipase; Enzyme Stability; Temperature; Enzymes, Immobilized; Rhizomucor
PubMed: 36912632
DOI: 10.1128/aem.02172-22 -
Journal of Fungi (Basel, Switzerland) Feb 2023Isavuconazole is the only US FDA-approved antifungal for treating invasive mucormycosis. We evaluated isavuconazole activity against a global collection of Mucorales...
Isavuconazole is the only US FDA-approved antifungal for treating invasive mucormycosis. We evaluated isavuconazole activity against a global collection of Mucorales isolates. Fifty-two isolates were collected during 2017-2020 from hospitals located in the USA, Europe, and the Asia-Pacific. Isolates were identified by MALDI-TOF MS and/or DNA sequencing and susceptibility tested by the broth microdilution method following CLSI guidelines. Isavuconazole (MIC, 2/>8 mg/L) inhibited 59.6% and 71.2% of all Mucorales isolates at ≤2 mg/L and ≤4 mg/L, respectively. Among comparators, amphotericin B (MIC, 0.5/1 mg/L) displayed the highest activity, followed by posaconazole (MIC, 0.5/8 mg/L). Voriconazole (MIC, >8/>8 mg/L) and the echinocandins (MIC, >4/>4 mg/L) had limited activity against Mucorales isolates. Isavuconazole activity varied by species and this agent inhibited at ≤4 mg/L 85.2%, 72.7%, and 25% of spp. ( = 27; MIC, 1/>8 mg/L), spp. ( = 11; MIC, 4/8 mg/L), and spp. ( = 8; MIC, >8 mg/L) isolates, respectively. Posaconazole MIC values against , , and species were 0.5/8 mg/L, 0.5/1 mg/L, and 2/- mg/L, respectively; amphotericin B MIC values were 1/1 mg/L, 0.5/1 mg/L, and 0.5/- mg/L, respectively. As susceptibility profiles varied among Mucorales genera, species identification and antifungal susceptibility testing are advised whenever possible to manage and monitor mucormycosis.
PubMed: 36836355
DOI: 10.3390/jof9020241 -
Journal of Fungi (Basel, Switzerland) Jan 2023Mucormycetes, a heterogeneous group of fungi, induce a life-threatening disease called mucormycosis. Immune deficiencies represent a major risk factor; hence, we wanted...
BACKGROUND
Mucormycetes, a heterogeneous group of fungi, induce a life-threatening disease called mucormycosis. Immune deficiencies represent a major risk factor; hence, we wanted to illuminate the role of complement and platelets in the defense against mucormycetes.
METHODS
(), (), (), (), (), and () spores were opsonized with human and mouse serum, and C1q, C3c, and terminal complement complex (C5b-9) deposition was measured. Additionally, thrombocytopenic, C3-deficient, or C6-deficient mice were intravenously infected with selected isolates. Survival and immunological parameters were monitored, and fungal burden was determined and compared to that of immunocompetent and neutropenic mice.
RESULTS
In vitro experiments showed significant differences in complement deposition between mucormycetes. isolates bound up to threefold more human C5b-9 than other mucormycetes. , , and bound high levels of murine C3c, whereas human C3c deposition was reduced on compared to and . Murine C3c deposition negatively correlated with virulence. Complement deficiencies and neutropenia, but not thrombocytopenia, were shown to be a risk factor for a lethal outcome.
CONCLUSION
Complement deposition varies between mucormycetes. Additionally, we demonstrated that complement and neutrophilic granulocytes, but not platelets, play an important role in a murine model of disseminated mucormycosis.
PubMed: 36836277
DOI: 10.3390/jof9020162 -
Indian Journal of Otolaryngology and... Jun 2023An invasive fungal infection caused by saprophytic and aerobic fungi Rhizopus, Rhizomucor, and Cunninghamella genera of the family Mucoraceae is known as Mucormycosis....
An invasive fungal infection caused by saprophytic and aerobic fungi Rhizopus, Rhizomucor, and Cunninghamella genera of the family Mucoraceae is known as Mucormycosis. Typically, Mucormycosis manifests in patients with conditions like uncontrolled diabetes, renal failure, patients receiving chemotherapy, long term steroid therapy or patients who are immunocompromised such as Acquired Immuno Deficiency Syndrome. The authors report a case of mixed opportunistic fungal infection of Mucormycosis and Aspergillosis following maxillofacial trauma that was treated by both medical and surgical line of management.
PubMed: 36777925
DOI: 10.1007/s12070-023-03492-z -
Foods (Basel, Switzerland) Jan 2023The fungal community in Qu plays a key role in the formation of turbid rice wine (TRW) style. The Sichuan Basin and its surrounding areas have become one of the main TRW...
The fungal community in Qu plays a key role in the formation of turbid rice wine (TRW) style. The Sichuan Basin and its surrounding areas have become one of the main TRW production regions in China; however, the fungal community in Qu and how they affect the characteristics of TRW remain unknown. Therefore, this study provided insight into the fungal biomarkers in Qu from Guang'an (GQ), Dazhou (DQ), Aba (AQ), and Liangshan (LQ), as well as their relationships with compounds in TRW. The main biomarkers in GQ were , , , and . However, they changed to and in DQ, in AQ, and and in LQ. As a response to fungal biomarkers, the reducing sugar, ethanol, organic acids, and volatile compounds were also changed markedly in TRWs. Among important volatile compounds (VIP > 1.00), phenethyl alcohol (14.1-29.4%) was dominant in TRWs. Meanwhile, 3-methyl-1-butanol (20.6-56.5%) was dominant in all TRWs except that fermented by GQ (GW). Acetic acid (29.4%) and ethyl palmitate (10.1%) were dominant in GW and LW, respectively. Moreover, GQ biomarkers were positively correlated with acetic acid and all unique important volatile compounds in GW. DQ biomarkers had positive correlations with unique compounds of acetoin and ethyl 5-chloro-1,3,4-thiadiazole-2-carboxylate in DW. Meanwhile, the AQ biomarkers were positively correlated with all AW unique, important, and volatile compounds. Although there were not any unique volatile compounds in LW, 16 important volatile compounds in LW were positively related to LQ biomarkers. Obviously, biomarkers in different geographic Qu played vital roles in the formation of important volatile compounds, which could contribute specific flavor to TRWs. This study provided a scientific understanding for future efforts to promote the excellent characteristics of TRW by regulating beneficial fungal communities.
PubMed: 36766114
DOI: 10.3390/foods12030585 -
Foods (Basel, Switzerland) Jan 2023In this study, the microbial diversity in Tibetan flavor was analyzed based on single molecule real-time sequencing (SMRT). The volatile flavor compounds in were...
In this study, the microbial diversity in Tibetan flavor was analyzed based on single molecule real-time sequencing (SMRT). The volatile flavor compounds in were detected using the headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). In addition, the correlation between various microbes and volatile flavor compounds was explored. Our studies indicated that the dominant bacterial genera in Tibetan flavor were , , , , , , , , and . The dominant fungal genera were , , and . 101 volatile compounds were detected in the samples, including alcohols, acids, esters, aldehydes, hydrocarbons, ketones, ethers, aromatics and pyrazines, and 10 key flavor compounds were identified using the relative odor activity value (ROAV). The results of our correlation analysis showed that was mainly associated with the synthesis of aldehydes such as trans-2-octenal, and was associated with the synthesis of most aldehydes. This paper has systematically investigated the physicochemical indices, microbial community structure and flavor compounds of Tibetan flavor , which is helpful in gaining a deeper understanding of the characteristics of Tibetan flavor .
PubMed: 36673416
DOI: 10.3390/foods12020324 -
Current Medical Mycology Jun 2022Invasive mucormycosis is a rare mycosis that affects most cases of uncontrolled diabetes and has a high mortality rate. Patients with COVID-19 are at high risk of...
BACKGROUND AND PURPOSE
Invasive mucormycosis is a rare mycosis that affects most cases of uncontrolled diabetes and has a high mortality rate. Patients with COVID-19 are at high risk of developing invasive mucormycosis due to the consumption of anti-inflammatory drugs such as corticosteroids and dexamethasone. species followed by spp. and spp. are the main common etiological agents of rhino-orbital mucormycosis. Therefore, this study aimed to present a case of mucormycosis due to in a diabetic patient with COVID-19 for the first time in Iran.
CASE REPORT
A 73-year-old diabetic female was referred to Ayatollah Rouhani Hospital in Babol, Iran, with a confirmed COVID-19 diagnosis, based on positive RT-PCR and computed tomography of the lungs. She has received methylprednisolone due to severe lung complications. Nasal involvement and left orbital swelling were observed 20 days after the hospitalization. By sinus endoscopic surgery, debridement was done and histopathology indicated wide hyphae (without septa). The sequenced PCR products displayed . In the antifungal susceptibility test, amphotericin B showed good activity against and the patient survived with timely treatment.
CONCLUSION
This is the first case report of rhino-orbital mucormycosis due to in COVID-19 patient; therefore, can be considered one of the etiological factors of rhino-orbital mucormycosis in COVID-19 cases.
PubMed: 36654791
DOI: 10.18502/cmm.8.2.10333 -
Infection and Drug Resistance 2022Metagenomic next-generation sequencing (mNGS) is a fast, sensitive and accurate diagnostic method for pathogens detection. However, reports on the application of mNGS in...
BACKGROUND
Metagenomic next-generation sequencing (mNGS) is a fast, sensitive and accurate diagnostic method for pathogens detection. However, reports on the application of mNGS in mucormycosis remain scarce.
METHODS
From January 2019 to December 2021, we recruited 13 patients with hematological malignancies who were suspected of mucormycosis and completed mNGS in D20. Then we retrospectively analyze the clinical data, diagnosis, therapeutic process, and outcomes. In order to evaluate the diagnostic value of mNGS in hematological malignancies patients with suspected mucormycosis.
RESULTS
All patients had high risk factors of Invasive Fungal Disease, including hematopoietic stem cell transplantation, immunosuppression, glucocorticoids, etc. The clinical presentations were pulmonary (n=9), rhino-orbito-cerebral (n=4). But the manifestations were nonspecific. All enrolled patients completed mNGS. And most (8/13, 61.54%) of the samples were from blood. Fungi can be detected in all specimens, including Rhizopus (n=7), Rhizomucor (n=4) and Mucor (n=2). In addition, 7/13 (53.85%) specimens were detected bacteria at the same time and virus were detected in 5/13 (38.46%). Histopathological examination was completed in 5 patients, 3 of which were completely consistent with the results of mNGS. After treatment, 6 patients were cured, while the other 7 patients died.
CONCLUSION
mNGS may be a complementary method for early diagnosis, especially for patients who are not suitable for histopathology examination or unable to obtain culture specimen. mNGS can also help detect bacteria and viruses simultaneously, allowing for appropriate and timely antibiotic administration and thus improving patient outcomes.
PubMed: 36570711
DOI: 10.2147/IDR.S393201