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This research aimed to develop a rapid and nondestructive method to model the growth and discrimination of spoilage fungi, like Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum, based on hyperspectral imaging system (HIS). A hyperspectral imaging system was used to measure the spectral response of fungi inoculated on potato dextrose agar plates and stored at 28°C and 85% RH. The fungi were analyzed every 12 h over two days during growth, and optimal simulation models were built based on HIS parameters. The results showed that the coefficients of determination (R2) of simulation models for testing datasets were 0.7223 to 0.9914, and the sum square error (SSE) and root mean square error (RMSE) were in a range of 2.03-53.40×10(-4) and 0.011-0.756, respectively. The correlation coefficients between the HIS parameters and colony forming units of fungi were high from 0.887 to 0.957. In addition, fungi species was discriminated by partial least squares discrimination analysis (PLSDA), with the classification accuracy of 97.5% for the test dataset at 36 h. The application of this method in real food has been addressed through the analysis of Botrytis cinerea, Rhizopus stolonifer and Colletotrichum acutatum inoculated in peaches, demonstrating that the HIS technique was effective for simulation of fungal infection in real food. This paper supplied a new technique and useful information for further study into modeling the growth of fungi and detecting fruit spoilage caused by fungi based on HIS.

In this study, we evaluated the efficacy of coatings comprising shrimp chitosan (CHI) and Mentha piperita L. (MPEO) or Mentha × villosa Huds (MVEO) essential oils to control mold infections caused by Aspergillus niger, Botrytis cinerea, Penicillium expansum and Rhizopus stolonifer in cherry tomato fruits (Solanum lycopersicum L.) during storage at room temperature (25°C for 12 days) and low temperature (12°C for 24 days). The effects of the coatings on the physicochemical and sensory characteristics of cherry tomato fruits during storage were also assessed. The minimum inhibitory concentration (MIC) of CHI against all test fungi was 8 mg/mL, whereas the MIC for both MPEO and MVEO was 5 μL/mL. Combinations of CHI at 4 mg/mL and MPEO or MVEO at 2.5 or 1.25 μL/mL strongly inhibited the mycelial growth and spore germination of target fungi. The coatings comprising CHI and MPEO or CHI and MVEO at the different tested concentrations delayed the growth of decay-causing fungi in artificially contaminated tomato fruit during storage at either room temperature or low temperature. The assayed coatings preserved the quality of cherry tomato fruit during storage, in terms of physicochemical and sensory attributes. These results indicate that coatings comprising CHI and MPEO or CHI and MVEO represent promising postharvest treatments to prevent common postharvest mold infections in cherry tomato fruit during storage without affecting the quality of the fruit., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Objectives: The aim of this study was to determine the susceptibility of 77 mould strains: Aspergillus fumigatus (20), Aspergillus flavus (8), Aspergillus niger (4), Aspergillus ochraceus (2), Penicillium citrinum (15), Penicillium crysogenum (14), Penicillium aurantiogriseum (1), Penicillium roquefortii (4), Penicillium paneum (2), Rhizopus spp. (3), Tricoderma spp. (1) and Mucor spp. (3) to biocides., Methods: MIC determination was determined based on CLSI methodology., Results: For hospital acquired strains, MIC50 was 0.5mg/L, MIC90 was 1mg/L for chlorhexidine (CHX); MIC50 was 0.5mg/L, and MIC90 was 1mg/L for benzalkonium chloride (BZC); MIC50 was 1mg/L, and MIC90 was 2mg/L for triclosan (TRC); MIC50 was 1024mg/L, and MIC90 was 2048mg/L for sodium hypochloride (SHC). For feed and food isolates MIC50 was 2mg/L, MIC90 was 8mg/L for CHX, MIC50 was 2mg/L, and MIC90 was 4mg/L for BZC, MIC50 was 2mg/L, and MIC90 was 4mg/L for TRC, MIC50 was 256mg/L, and MIC90 was 512mg/L for SHC., Conclusion: We can conclude that food isolates presented slightly higher MIC50 and MIC90 values for CHX, BNZ and TRC, but not for SHC., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Triazole prophylaxis has become the norm in patients with hematological malignancies. Breakthrough infections caused by Mucorales during triazole prophylaxis remain a challenging problem. We found that preexposure of Rhizopus oryzae to antifungal triazoles (fluconazole, voriconazole, posaconazole, and itraconazole) did not modify the in vitro susceptibility of Rhizopus oryzae to posaconazole. In contrast, preexposure of Rhizopus to triazoles was associated with the enhanced in vitro susceptibility of R. oryzae to amphotericin B. Preexposure to posaconazole did not alter the virulence of R. oryzae in the fly model of mucormycosis., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

The present research account deals with the bioproduction of fumaric acid (FA) from apple pomace ultrafiltration sludge (APUS) and apple pomace (AP) through fermentation. The filamentous fungus Rhizopus oryzae 1526 was used as a biocatalyst and its morphological impact on FA production was analysed in detail. For submerged fermentation, 40 g L(-1) of total solids concentration of APUS, pH 6.0, 30 °C, 200 rpm flask shaking speed and 72 h of incubation were found to be optimum for FA production (25.2 ± 1.0 g L(-1), 0.350 g (L(-1) h(-1))). Broth viscosity (cP), residual reducing sugar (g L(-1)) and ethanol (g L(-1)) produced as by-product, were also analysed. Plastic trays were used for solid state fermentation and at optimized level of moisture and incubation period, 52 ± 2.67 g FA per kg dry weight of AP was obtained. Changes in the total phenolic content (mg g(-1) dry weight of AP) were monitored at regular intervals. Utilization of APUS and AP for the directed synthesis of the high-value platform chemical FA by the fungal strain R. oryzae 1526 was an excellent display of fungal physiological and morphological control over a fermentative product., (Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

We studied the efficacy of the investigational drug VT-1161 against mucormycosis. VT-1161 had more potent in vitro activity against Rhizopus arrhizus var. arrhizus than against R. arrhizus var. delemar. VT-1161 treatment demonstrated dose-dependent plasma drug levels with prolonged survival time and lowered tissue fungal burden in immunosuppressed mice infected with R. arrhizus var. arrhizus and was as effective as high-dose liposomal amphotericin B treatment. These results support further development of VT-1161 against mucormycosis., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Rhizopus oryzae is strictly inhibited by biodiesel-based by-product crude glycerol, which results in low fumaric acid production. In this study, evolutionary engineering was employed to activate the glycerol utilization pathway for fumaric acid production. An evolved strain G80 was selected, which could tolerate and utilize high concentrations of crude glycerol to produce 14.9g/L fumaric acid with a yield of 0.248g/g glycerol. Key enzymes activity analysis revealed that the evolved strain displayed a significant upregulation in glycerol dissimilation, pyruvate consumption and reductive tricarboxylic acid pathways, compared with the parent strain. Subsequently, intracellular metabolic profiling analysis showed that amino acid biosynthesis, tricarboxylic acid cycle, fatty acid and stress response metabolites accounted for metabolic difference between two strains. Moreover, a glycerol fed-batch strategy was optimized to obtain the highest fumaric acid production of 25.5g/L, significantly increased by 20.9-fold than that of the parent strain of 1.2g/L., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

A mycelial mattress of Rhizopus stolonifer obtained from a liquid static culture was utilized for wound dressing and biomedical use. Following screening of mutants induced by UV radiation, F6, exhibiting delayed sporangium formation was selected because its sporangium maturation exhibited a 5-day delay without significant loss of mycelial weight compared to the wild type. The sporangium-free mycelial mattress from the sporangiospore culture of F6 was treated with 1N sodium hydroxide NaOH at 85°C for 2 h to produce a sponge-like membrane named Rhizochitin. The trifluoroacetic acid hydrolysate of Rhizochitin contained 36% N-acetylglucosamine and 53% hexose respectively detected by the Elson-Morgen and phenol-sulfuric acid methods. Results indicated the wound area in rats covered with Rhizochitin was 40% less than that of the uncovered group. Rhizochitin decreased the expression of PDGF in the proliferation stage, increased the expression of TGF-β in the inflammation and proliferation stages, and increased the expression of VEGF in the inflammation and proliferation stages. Rhizochitin inhibited secretion of matrix metalloproteinase-9 on days 1, 7, 9, and 12 and matrix metalloproteinase-2 on days 3, 7, 9, and 12. It was concluded that Rhizochitin has beneficial properties of biocompatible, biodegradable, and wound healing.

Background: Tempeh is a food product obtained from legumes by means of solid-state fermentation with Rhizopus sp. Our previous research proved that mixed-culture inoculum may also be successfully applied. The objective of present research was to study the proteolytic activity of R. microsporus var. chinensis and A. oryzae during tempeh-type fermentation of grass pea seeds, and the effect of inoculum composition on the protein level and in vitro protein bioavailability in products., Methods: Fermentation substrate were soaked and cooked grass pea seeds. Material was mixed with single- or mixed-culture inoculum, and incubated in perforated plastic bags at 30°C for 32 hrs. In the products, the proteolytic activity (pH 3, 5 and 7), glucosamine, total protein and free amino acids levels, as well as protein in vitro bioavailability and degree of protein hydrolysis were obtained., Results: The significant correlation was found between glucosamine content and proteolytic activity in grass pea seeds fermented with Rhizopus or Aspergillus. The activities of Rhizopus proteases were higher than Aspergillus ones, which corresponded with the degree of seed protein hydrolysis. Both strains showed the highest activity of protease at pH 3. Tempeh made with pure culture of Rhizopus had 37% protein of 69% in-vitro bioavailability. Mixed-culture fermentation improved nutritional parameters of products only when the dose of Aspergillus spores in the inoculum was equal and lower than that of Rhizopus. This process resulted in higher in-vitro bioavailability of protein, slightly more efficient protein hydrolysis and higher level of free amino acids, as compared to standard tempeh., Conclusions: The activity of A. oryzae in tempeh-type fermentation is beneficial as long as it does not dominate the activity and/or growth of Rhizopus strain.

Current models that describe the extension of fungal hyphae and development of a mycelium either do not describe the role of vesicles in hyphal extension or do not correctly describe the experimentally observed profile for distribution of vesicles along the hypha. The present work uses the n-tanks-in-series approach to develop a model for hyphal extension that describes the intracellular transport of nutrient to a sub-apical zone where vesicles are formed and then transported to the tip, where tip extension occurs. The model was calibrated using experimental data from the literature for the extension of reproductive aerial hyphae of three different fungi, and was able to describe different profiles involving acceleration and deceleration of the extension rate. A sensitivity analysis showed that the supply of nutrient to the sub-apical vesicle-producing zone is a key factor influencing the rate of extension of the hypha. Although this model was used to describe the extension of a single reproductive aerial hypha, the use of the n-tanks-in-series approach to representing the hypha means that the model has the flexibility to be extended to describe the growth of other types of hyphae and the branching of hyphae to form a complete mycelium.

Glucocorticoid (GC) use is a common risk factor for invasive fungal infections. This is attributed to the complex dysregulation of immunity caused by GCs. However, studies have demonstrated increased growth with GC exposure for some molds, such as Aspergillus fumigatus and Exserohilum rostratum. No such data exist for Mucorales. Therefore, we investigated the influence of GC exposure on the growth of Rhizopus arrhizus (syn. R. oryzae) in different culture media and in different atmospheres. We measured continuous spore growth using spectrophotometry and biomass variations using XTT assay. We did not observe enhanced growth or biomass variation with any of the GCs regardless of the medium or conditions. These results support the existence of fungus-specific differences in the effect of GCs on fungal biology.

The in vitro antifungal activities and mechanism of action of tea polyphenols (TP), tea saponin (TS) and their combination were evaluated against Rhizopus stolonifer. The results showed that both TP and TS inhibited the mycelial growth in a dose-dependent manner, and their combination at the ratio of 7:3 exhibited synergistic antifungal interaction. We also observed that the treatment of TP or TS significantly induced the production of H2O2 and resulted in membrane lipid peroxidation, thus leading to an increase in cell membrane permeability and the leakage of K(+), soluble protein and soluble sugar. Moreover, combining them for treatment increased the induction of H2O2 production and oxidative damage. Scanning electron microscopic observations also showed the damage to the hyphal cell structure. It was concluded that TP, TS and their combination inhibit the growth of R. stolonifer through the induction of H2O2 production, leading to cell membrane oxidative damage and intracellular constituent leakage. These findings suggest that TP and TS can potentially be used as an alternative to control postharvest fruit diseases caused by R. stolonifer., (© 2015 S. Karger AG, Basel.)

Mucormycoses are fungal infections caused by the ancient Mucorales. They are rare, but increasingly reported. Predisposing conditions supporting and favoring mucormycoses in humans and animals include diabetic ketoacidosis, immunosuppression and haematological malignancies. However, comprehensive surveys to elucidate fungal virulence in ancient fungi are limited and so far focused on Lichtheimia and Mucor. The presented study focused on one of the most important causative agent of mucormycoses, the genus Rhizopus (Rhizopodaceae). All known clinically-relevant species are thermotolerant and are monophyletic. They are more virulent compared to non-clinically, mesophilic species. Although adaptation to elevated temperatures correlated with the virulence of the species, mesophilic strains showed also lower virulence in Galleria mellonella incubated at permissive temperatures indicating the existence of additional factors involved in the pathogenesis of clinical Rhizopus species. However, neither specific adaptation to nutritional requirements nor stress resistance correlated with virulence, supporting the idea that Mucorales are predominantly saprotrophs without a specific adaptation to warm blooded hosts.

The aim of this study was to extract chitosan (CHI) from Mucor circinelloides UCP 050 grown in a corn steep liquor (CSL)-based medium under optimized conditions and to assess the efficacy of the obtained CHI to inhibit the post-harvest pathogenic fungi Aspergillus niger URM 5162 and Rhizopus stolonifer URM 3482 in laboratory media and as a coating on table grapes (Vitis labrusca L.). The effect of CHI coating on some physical, physicochemical and sensory characteristics of the fruits during storage was assessed. The greatest amount of CHI was extracted from M. circinelloides UCP 050 grown in medium containing 7 g of CSL per 100 mL at pH 5.5 with rotation at 180 rpm. CHI from M. circinelloides UCP 050 caused morphological changes in the spores of the fungal strains tested and inhibited mycelial growth and spore germination. CHI coating delayed the growth of the assayed fungal strains in artificially infected grapes, as well as autochthonous mycoflora during storage. CHI coating preserved the quality of grapes during storage, as measured by their physical, physicochemical and sensory attributes. These results demonstrate that edible coatings derived from M. circinelloides CHI could be a useful alternative for controlling pathogenic fungi and maintaining the post-harvest quality of table grapes., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Mucormycosis has emerged as a relatively common severe mycosis in patients with haematological and allogeneic stem cell transplantation. Source of transmission is from unidentified sources in the environment. Early diagnosis of infection and its source of contamination are paramount for rapid and appropriate therapy. In this study, rolling circle amplification (RCA) is introduced as a sensitive, specific and reproducible isothermal DNA amplification technique for rapid molecular identification of six of the most virulent species (Rhizopus microsporus, R. arrhizus var. arrhizus, R. arrhizus var. delemar, Mucor irregularis, Mucor circinelloides, Lichtheimia ramosa, Lichtheimia corymbifera). DNAs of target species were successfully amplified, with no cross reactivity between species. RCA can be considered as a rapid detection method with high specificity and sensitivity, suitable for large screening., (© 2014 Blackwell Verlag GmbH.)

Background: Fungal contamination of poultry feed causes economic losses to industry and represents a potential risk to animal health. The aim of the present study was to analyze the effectiveness of whey fermented with kefir grains as additive to reduce fungal incidence, thus improving feed safety., Results: Whey fermented for 24 h at 20 °C with kefir grains (100 g L(-1) ) reduced conidial germination of Aspergillus flavus, Aspergillus parasiticus, Aspergillus terreus, Aspergillus fumigatus, Penicillium crustosum, Trichoderma longibrachiatum and Rhizopus sp. Poultry feed supplemented with fermented whey (1 L kg(-1) ) was two to four times more resistant to fungal contamination than control feed depending on the fungal species. Additionally, it contained kefir microorganisms at levels of 1 × 10(8) colony-forming units (CFU) kg(-1) of lactic acid bacteria and 6 × 10(7) CFU kg(-1) of yeasts even after 30 days of storage., Conclusion: Fermented whey added to poultry feed acted as a biopreservative, improving its resistance to fungal contamination and increasing its shelf life., (© 2014 Society of Chemical Industry.)

The effects of pellet morphology, diameter, density, and interior structure on L-lactic acid fermentation by Rhizopus oryzae were characterized for different inoculum sizes and concentrations of peptone and CaCO3. Inoculum size was the most important factor determining pellet formation and diameter. The diameter decreased with increasing inoculum size, and larger pellets were observed for lower inoculum sizes. Peptone concentration had the greatest effect on pellet density, which increased with increasing peptone concentration. L-lactic acid production depended heavily on pellet density but not on pellet diameter. Low-density pellets formed easily under conditions of low peptone concentration and often had a relatively hollow structure, with a thin condensed layer surrounding the pellet and an extraordinarily loose biomass or hollow center. As expected, this structure greatly decreased production. The production of L-lactic acid increased until the density reached a certain level (50-60 kg/m(3)), in which the compact part was distributed homogeneously in the thick outer layer of the pellet and loose in the central layer. Homogeneously structured, denser pellets had limited mass transfer, causing a lower overall turnover rate. However, the interior structure remained nearly constant throughout all fermentation phases for pellets with the same density. CaCO3 concentration only had a slight influence on pellet diameter and density, probably because it increases spore germination and filamentous hypha extension. This work also provides a new analysis method to quantify the interior structure of pellets, thus giving insight into pellet structure and its relationship with productivity.

Zearalenone (ZEN) is a nonsteroidal estrogenic mycotoxin biosynthesized by various Fusarium fungi. These fungal species frequently infest grains; therefore, ZEN represents a common contaminant in cereal products. The biotransformation of ZEN differs significantly from species to species, and several metabolites are known to be formed by animals, plants, and microorganisms. The aim of the present study was to investigate the microbial conversion of ZEN by species of the genera Rhizopus and Aspergillus representing relevant fungi for food processing (e.g. fermentation). To monitor the ZEN metabolism, ZEN was added to liquid cultures of the different fungal species. After a period of 3 days, the media were analyzed by HPLC-MS/MS for metabolite formation. Two Aspergillus oryzae strains and all seven Rhizopus species were able to convert ZEN into various metabolites, including ZEN-14-sulfate as well as ZEN-O-14- and ZEN-O-16-glucoside. Microbial transformation of ZEN into the significantly more estrogenic α-zearalenol (α-ZEL) was also observed. Additionally, a novel fungal metabolite, α-ZEL-sulfate, was detected. Semi-quantification of the main metabolites indicates that more than 50% of initial ZEN may be modified. The results show that fungal strains have the potential to convert ZEN into various metabolites leading to a masking of the toxin, for example in fermented food., (© 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.)

Unlabelled: The kinetics of cultivation of zygomycete filamentous fungus Rhizopus oryzae in deproteinized whey medium in relation to chitosan production was studied here to optimize chitosan production from R. oryzae as well as utilize whey, a by-product of sweetmeat industry as a cheap source of sugar in the cultivation process. Chitosan content of R. oryzae biomass was found to be increased with time during cultivation and reached maximum (13·6%) after 72 h and then declined steadily. Maximum 1·13 g of chitosan was obtained from one litre of deproteinized whey medium. Concentration of lactose in the medium was observed to be reduced from 45·0 to 11·7 g l(-1) during cultivation resulting in decrease in biochemical oxygen demand (BOD) of whey by approx. 60%, and this was important from environmental point of view before discharging whey into any water body. However, no significant change in pH or titratable acidity was noted during the entire course of cultivation, probably due to good buffering capacity of the medium. Molecular weight of chitosan varied from 130 to 230 kDa depending on the time of cultivation, but no significant change in degree of deacetylation of chitosan (approx. 87%) was found during cultivation., Significance and Impact of the Study: Whey is the largest by-product of dairy industries, and its disposal is a big environmental issue because of its high biological oxygen demand (BOD) value. This study will help to lower BOD value of whey by using it as a cultivation medium for fungus R. oryzae that contains chitosan, a very commercially important material on its cell wall. Moreover, the study on biochemical changes in whey during cultivation process with R. oryzae will help to understand the exact changes occurring in the medium and optimize cultivation process to isolate chitosan in larger extent with better and uniform physicochemical properties., (© 2014 The Society for Applied Microbiology.)

The antifungal activities of chitosan and oligochitosan have been used to control postharvest decay of the fruits. The effect of chitosan and oligochitosan on mycelium growth, spore germination, and mitochondrial function of Rhizopus stolonifer was evaluated in order to establish a connection between fungus development and the main organelle in charge to provide energy to the cell. The mycelium growth of R. stolonifer was significantly reduced on minimum media amended with chitosan or oligochitosan. The highest antifungal indexes were obtained on media containing chitosan or oligochitosan at 2.0 mg ml(-1). Microscopic observation showed that chitosan and oligochitosan affected the spore germination and hyphae morphology. Both polymers increased oxygen consumption of R. stolonifer. Respiratory activity was restored with NADH in permeabilized treated and untreated cells, and was inhibited with rotenone and flavones. Complex III and IV were inhibited by antimycin A and cyanide, respectively, in treated and untreated cells. Chitosan and oligochitosan increased NADH dehydrogenase activity in isolated mitochondria. However, there were not changes in the cytochrome c oxidase and ATPase activities by effect of these polymers. These results suggest that both chitosan and oligochitosan affect the development of R. stolonifer and might be implicated in the mitochondrial dysfunction., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A complex bioflocculant MBF917 was prepared by Rhizopus sp. M9 and M17 using potato starch wastewater (PSW) as nutrilite, and its flocculation characteristics of treating PSW were studied. Culture conditions of the two strains were optimized, and flocculating conditions of the bioflocculant for treating PSW were also investigated. The optimal and economical culture conditions were determined as COD of about 1600 mg/L, 0.3 g/L urea and 0.04 g/L potassium dihydrogen phosphate, with no need of adding carbon sources or adjusting pH. When the bioflocculant was used to flocculate PSW, the optimal dosage was 0.1 mL/L with addition of 5 mL/L 10% CaCl2 as coagulant aid, and there was no need to adjust pH. After flocculation, COD and turbidity removal rates of the PSW could reach 54.09% and 92.11%, respectively, and 1.1g/L proteic substance was recycled from the PSW as a byproduct that could be used for animal feed., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Five microorganisms, namely Aspergillus niger CECT 2700, A. niger CECT 2915, A. niger CECT 2088, Aspergillus terreus CECT 2808, and Rhizopus stolonifer CECT 2344, were grown on corncob to produce cell wall polysaccharide-degrading enzymes, mainly xylanases, by solid-state fermentation (SSF). A. niger CECT 2700 produced the highest amount of xylanases of 504 ± 7 U/g dry corncob (dcc) after 3 days of fermentation. The optimization of the culture broth (5.0 g/L NaNO3, 1.3 g/L (NH4)2SO4, 4.5 g/L KH2PO4, and 3 g/L yeast extract) and operational conditions (5 g of bed loading, using an initial substrate to moistening medium of 1:3.6 (w/v)) allowed increasing the predicted maximal xylanase activity up to 2,452.7 U/g dcc. However, different pretreatments of materials, including destarching, autoclaving, microwave, and alkaline treatments, were detrimental. Finally, the process was successfully established in a laboratory-scale horizontal tube bioreactor, achieving the highest xylanase activity (2,926 U/g dcc) at a flow rate of 0.2 L/min. The result showed an overall 5.8-fold increase in xylanase activity after optimization of culture media, operational conditions, and scale-up.

The leaves and bark of Croton cajucara, a shrub from the Amazon region, have been used in folk medicine to treat diabetes, malaria, and gastrointestinal and liver disorders. The essential oil from the leaves, rich in linalool, presented antileishmanial and antimicrobial activities. A chemotype of this species was found with an essential oil rich in 7-hydroxycalamenene. During our studies of the C. cajucara essential oil, we isolated 7-hydroxycalamenene at > 98 % purity. The minimum inhibitory concentration of 7-hydroxycalamenene against Absidia cylindrospora, Cunninghamella elegans, Mucor circinelloides, Mucor circinelloides f. circinelloides, Mucor mucedo, Mucor plumbeus, Mucor ramosissimus, Rhizopus microsporus, Rhizopus oryzae, and Syncephalastrum racemosum ranged from 19.53 to 2500 µg/mL. The reference drug used, amphotericin B, presented a minimum inhibitory concentration ranging from 0.085 µg/mL to 43.87 µg/mL. 7-Hydroxycalamenene also altered spore differentiation and total lipid content. Ultrastructural analysis by transmission electron microscopy showed significant alterations in the cellular structure of R. oryzae., (Georg Thieme Verlag KG Stuttgart · New York.)

Rhizopus oligosporus was cultivated on thin stillage from a dry-grind corn ethanol plant. The aim of the research was to develop a process to replace the current energy-intensive flash evaporation and make use of this nutrient-rich stream to create a new co-product in the form of protein-rich biomass. Batch experiments in 5- and 50-L stirred bioreactors showed prolific fungal growth under non-sterile conditions. COD, suspended solids, glycerol, and organic acids removals, critical for in-plant water reuse, reached ca. 80%, 98%, 100% and 100%, respectively, within 5 d of fungal inoculation, enabling effluent recycle as process water. R. oligosporus contains 2% lysine, good levels of other essential amino acids, and 43% crude protein - a highly nutritious livestock feed. Avoiding water evaporation from thin stillage would furthermore save substantial energy inputs on corn ethanol plants., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Plant extracts appear to be one of the most effective alternative methods of plant diseases control which are less harmful to human beings and environment. In vitro antifungal activity of methanolic extracts of three promising wild geophytic plants against three post-harvest pathogenic fungi using radial growth technique was conducted. These extracts included the shoot system (S) and underground parts (R) of Asparagus stipularis, Cyperus capitatus and Stipagrostis lanata. The tested fungi were Alternaria solani, Aspergillus niger and Rhizopus stolonifer. The results exhibited that, all plant extracts had antifungal activity against the tested fungi. The antifungal activity greatly varied depending on plant parts and/or plant species. R. stolonifer was the most susceptible fungus to the tested plant extracts followed by A. niger and then A. solani. On the other hand, the most effective plant extracts against tested fungi were S. lanata (S) and A. stipularis (R). The most effective plant extracts against R. stolonifer were S. lanata (R) and C. capitatus (S). While, the extracts of A. stipularis (R) and S. lanata (S) were the most effective against A. niger. The extracts of C. capitatus (S) and S. lanata (S) exhibited the highest antifungal activity against A. solani. The results demonstrated that, the methanolic extracts of A. stipularis, C. capitatus and S. lanata had potential antifungal activity against A. solani, A. niger and R. stolonifer.

We characterized the antifungal activity of the Bacillus circulans subclass III MH-K1 chitosanase (MH-K1 chitosanase), which is one of the most intensively studied glycoside hydrolases (GHs) that belong to GH family 46. MH-K1 chitosanase inhibited the growth of zygomycetes fungi, Rhizopus and Mucor, even at 10 pmol (0.3 μg)/ml culture probably via its fungistatic effect. The amino acid substitution E37Q abolished the antifungal activity of MH-K1 chitosanase, but retained binding to chitotriose. The E37Q mutant was fused with green fluorescent protein (GFP) at its N-terminus and proved to act as a chitosan probe in combination with wheat-germ agglutinin (WGA), which is a chitin-specific binding lectin. The GFP-fused MH-K1 chitosanase mutant E37Q (GFP-E37Q) bound clearly to the hyphae of the Rhizopus and Mucor strains, indicating the presence of chitosan. In contrast, Cy5-labelled WGA (Cy5-WGA), but not GFP-E37Q, stained the hyphae of non-zygomycetes species, i.e. Fusarium oxysporum, Penicillium expansum, and Aspergillus awamori. When the mycelia of Rhizopus oryzae were treated with wild type MH-K1 chitosanase, they could not bind to GFP-E37Q but were stained instead by Cy5-WGA. We conclude that chitin is covered by chitosan in the cell walls of R. oryzae.

The morphological effects of an antifungal activity produced by Lactobacillus plantarum 16, a malt-production steep water isolate, on two food-associated fungi were examined microscopically. Spore germination was completely inhibited in Aspergillus fumigatus and Rhizopus stolonifer upon treatment with concentrated cell-free supernatant (cCFS) of strain 16. Furthermore, addition of antifungal cCFS to germ tubes and hyphae halted further development compared to untreated controls. Transcriptome analysis of A. fumigatus Af293 following exposure to antifungal cCFS revealed a number of genes with altered transcription involved in a variety of cellular functions, most notably cell metabolism, suggesting a global metabolic shutdown and subsequent cell death. Increased transcription of the global regulator LaeA was also observed indicating that exposure to the antifungal activity caused a cellular stress response.

The high mortality rate of mucormycosis with currently available monotherapy has created interest in studying novel strategies for antifungal agents. With the exception of amphotericin B (AMB), the triazoles (posaconazole [PCZ] and itraconazole [ICZ]) are fungistatic in vitro against Rhizopus oryzae . We hypothesized that growth at a high temperature (42°C) results in fungicidal activity of PCZ and ICZ that is mediated through apoptosis. R. oryzae had high MIC values for PCZ and ICZ (16 to 64 μg/ml) at 25°C; in contrast, the MICs for PCZ and ICZ were significantly lower at 37°C (8 to 16 μg/ml) and 42°C (0.25 to 1 μg/ml). Furthermore, PCZ and ICZ dose-dependent inhibition of germination was more pronounced at 42°C than at 37°C. In addition, intracellular reactive oxygen species (ROS) increased significantly when fungi were exposed to antifungals at 42°C. Characteristic cellular changes of apoptosis in R. oryzae were induced by the accumulation of intracellular reactive oxygen species. Cells treated with PCZ or ICZ in combination with hyperthermia (42°C) exhibited characteristic markers of early apoptosis: phosphatidylserine externalization visualized by annexin V staining, membrane depolarization visualized by bis-[1,3-dibutylbarbituric acid] trimethine oxonol (DiBAC) staining, and increased metacaspase activity. Moreover, terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and DAPI (4',6-diamidino-2-phenylindole) staining demonstrated DNA fragmentation and condensation, respectively. The addition of N-acetylcysteine increased fungal survival, prevented apoptosis, reduced ROS accumulation, and decreased metacaspase activation. We concluded that hyperthermia, either alone or in the presence of PCZ or ICZ, induces apoptosis in R. oryzae. Local thermal delivery could be a therapeutically useful adjunct strategy for these refractory infections.

The efficacy of Bacillus subtilis SM21 on controlling Rhizopus rot caused by Rhizopus stolonifer in postharvest peach fruit and the possible mechanisms were investigated. The results indicated B. subtilis SM21 treatment reduced lesion diameter and disease incidence by 37.2% and 26.7% on the 2nd day of inoculation compared with the control. The in vitro test showed significant inhibitory effect of B. subtilis SM21 on mycelial growth of R. stolonifer with an inhibition rate of 48.9%. B. subtilis SM21 treatment significantly enhanced activities of chitinase and β-1,3-glucanase, and promoted accumulation of H2O2. Total phenolic content and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity were also increased by this treatment. Transcription of seven defense related genes was much stronger in fruit treated with B. subtilis SM21 or those both treated with B. subtilis SM21 and inoculated with R. stolonifer compared with fruit inoculated with R. stolonifer alone. These results suggest that B. subtilis SM21 can effectively inhibit Rhizopus rot caused by R. stolonifer in postharvest peach fruit, possibly by directly inhibiting growth of the pathogen, and indirectly inducing disease resistance in the fruit., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Rhizopus oligosporus has proven beneficial in the detoxification of lupin seeds. The fermentation process is mainly affected by the initial pH in the medium. In the range of growth of mold, there are maximum enzymatic activities in pH of 3.5 and 5.5. Metabolism change occurs at these pH levels; therefore, we studied the growth, pH changes, dry matter intake, and alkaloid degradation within 48 h of fermentation. Cultures of lupin agar (LA) with pH of 3.5 and 5.5 were made in Petri dishes with lupin flour. Results showed pH directly affects the degradation of alkaloids and fungal growth. Detoxification levels achieved were 16.58 and 63.23 % in treatments LA 3.5 and LA 5.5, respectively. Fungal growth was 0.919 mg/cm(2) in LA 3.5 and 1.081 mg/cm(2) in LA 5.5. Maximum degradation rate in LA 5.5 was given between 16 and 20 h, which coincided with maximum fungal growth. Despite having similar dry matter intake in both treatments, a pH of 3.5 did not show the same degree of detoxification. The analysis with exponential, yield of growth, yield of dry matter intake and luedeking and piret equations, confirm the relation between intake and growth with detoxification. Dry matter intake equation predicts with R (2) of 0.94 the detoxification in LA 5.5. A pH of 5.5 is directly related with detoxification and fungal development.

Thirty four patients with mucormycosis in 10 hospitals of St. Petersburg were observed in 2004-2013. The most frequent underlying diseases of mucormycosis were acute leukoses (64%). In 100% of the patients mucormycosis developed as a nosocomial infection. The risk factors, etiology, basic clinical signs and strategy in the treatment of mucormycosis were analyzed.

Mucormycoses are emerging infections that have high rates of morbidity and mortality. They show high resistance to antifungal agents, and there is a limited therapeutic arsenal currently available, therefore, there is a great need to give priority to testing therapeutic agents for the treatment of mucormycosis. Along this line, the use of essential oils and phytoconstituents has been emphasized as a new therapeutic approach. The objective of this work was to investigate the antifungal activity of the essential oil (EO) of Thymus vulgaris, and its constituents thymol and p-cymene against Rhizopus oryzae, through microbiological screening, determination of minimal inhibitory concentration (MICs) and minimal fungicidal concentration (MFCs), effects on mycelial growth and germination of sporangiospores and interaction with ergosterol. The MIC of EO and thymol varied 128-512 µg/mL, but the MFC of EO and thymol varied 512-1024 µg/mL and 128-1024 µg/mL, respectively. The results also showed that EO and thymol significantly inhibited mycelial development and germination of sporangiospores. Investigation of the mechanism of antifungal action showed that EO and thymol interact with ergosterol. These data indicate that EO of T. vulgaris and thymol possess strong antifungal activity, which can be related to their interaction with ergosterol, supporting the possible use of these products in the treatment of mucormycosis.

This study evaluated the efficacy of the combined application of chitosan (CHI) and Origanum vulgare L. essential oil (OV) in the inhibition of Rhizopus stolonifer URM 3728 and Aspergillus niger URM 5842 on laboratory media and on grapes (Vitis labrusca L.) and its influence on the physical, physicochemical and sensory characteristics of the fruits during storage (25 °C, 12 days and 12 °C, 24 days). The application of mixtures of different CHI and OV concentrations (Minimum Inhibitory Concentration - MIC, 1/2 MIC and 1/4 MIC) inhibited the mycelial growth of the test fungi. The application of CHI and OV at sub-inhibitory concentrations (CHI 1/2 MIC + OV 1/4 MIC; CHI 1/2 MIC + OV 1/2 MIC) inhibited spore germination and caused morphological changes in fungal spores and mycelia, in addition to inhibiting the growth of the assayed fungi strains in artificially infected grapes as well as the autochthonous mycoflora of grapes stored at both room and cold temperature. In general, the application of a coating composed of CHI and OV at sub-inhibitory concentrations preserved the quality of grapes as measured by their physical and physicochemical attributes, while some of their sensory attributes improved throughout the assessed storage time. These results demonstrate the potential of the combination of CHI and OV at sub-inhibitory concentrations to control post-harvest pathogenic fungi in fruits, in particular, R. stolonifer and A. niger in grapes., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

This study shows that Rhizopus oryzae is capable of directly utilizing cassava pulp alone to L-lactic acid in solid state fermentation (SSF). pH control at 6.0 helped prevent end product inhibition. Increasing lactate titer was observed at the higher initial moistened water due to the higher degree of substrate swelling and hydrolysis. With shaking, limited ethanol production but no change in lactate titer was observed. Rigorous shaking gave better oxygen transfer but presumably caused cell damage leading to substrate utilization through the biosynthesis route. Supplementing cassava pulp with nitrogen enhanced growth but not lactate production. Under the optimal conditions, R. oryzae converted the sole cassava pulp into lactic acid at the titer of 206.20 mg per g initial dry pulp. With the help of commercial cellulase and glucoamylase, the dramatically increasing lactate titer of 463.18 mg per g initial dry pulp was achieved via SSF.

Fumaric acid, a dicarboxylic acid used as a food acidulant and in manufacturing synthetic resins, can be produced from glucose in fermentation by Rhizopus oryzae. However, the fumaric acid yield is limited by the co-production of ethanol and other byproducts. To increase fumaric acid production, overexpressing endogenous pyruvate carboxylase (PYC) and exogenous phosphoenolpyruvate carboxylase (PEPC) to increase the carbon flux toward oxaloacetate were investigated. Compared to the wild type, the PYC activity in the pyc transformants increased 56%-83%, whereas pepc transformants exhibited significant PEPC activity (3-6 mU/mg) that was absent in the wild type. Fumaric acid production by the pepc transformant increased 26% (0.78 g/g glucose vs. 0.62 g/g for the wild type). However, the pyc transformants grew poorly and had low fumaric acid yields (<0.05 g/g glucose) due to the formation of large cell pellets that limited oxygen supply and resulted in the accumulation of ethanol with a high yield of 0.13-0.36 g/g glucose. This study is the first attempt to use metabolic engineering to modify the fumaric acid biosynthesis pathway to increase fumaric acid production in R. oryzae., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The antimicrobial activity of oregano has been attributed mainly to the presence of volatile compounds found in its essential oil (EO), mainly carvacrol and thymol. The search for antimicrobial activity of oregano EO with different concentrations of thymol and carvacrol, can lead to products with a wider range of applications. The aim of this work was to describe the in vitro antifungal effect of Mexican oregano (Lippia berlandieri Schauer) EO fractions on the growth of Aspergillus, Penicillium, and Rhizopus sp. The Mexican oregano EO fractions studied had different concentrations of carvacrol, which decreased from fraction 1 to 5 (81% to 23%), while thymol content increased from 3% to 64%. Fungal inhibition was evaluated on a wheat flour-based medium with EO fractions concentrations ranging from 50 to 200 mg/kg. Radial growth curves were fitted using the modified Gompertz model (R(2)(adj) = 0.989 ± 0.01). No significant differences (P > 0.05) were found with the different composition of the Mexican oregano EO fractions; nevertheless, fraction concentration presented significant (P < 0.05) mold inhibition as concentration increased. Rhizopus sp. (Rh18) showed a linear reduction on specific growth rate, on the maximum mold growth at the stationary phase, and an increase in the lag time as the concentration of the oregano EO increased; mold growth inhibition were achieved at 150 mg/kg in fractions 1 to 4, and at 100 mg/kg for fraction 5. Aspergillus sp. (As6) and Penicillium sp. (Pe36) were inhibited at 150 and 200 mg/kg, respectively. Results obtained suggest that Mexican oregano EO (Lippia berlandieri Schauer) compounds could be used as antimicrobial agents to prevent fungal growth in bakery products., (© 2012 Institute of Food Technologists®)

In recent years, there has been significant progress in the biological synthesis of nanomaterials. However, the molecular mechanism of gold biomineralization in microorganisms of industrial relevance remains largely unexplored. Here we describe the biosynthesis mechanism of gold nanoparticles (AuNPs) in the fungus Rhizopus oryzae . Reduction of AuCl(4)(-) [Au(III)] to nanoparticulate Au(0) (AuNPs) occurs in both the cell wall and cytoplasmic region of R. oryzae . The average size of the as-synthesized AuNPs is ~15 nm. The biomineralization occurs through adsorption, initial reduction to Au(I), followed by complexation [Au(I) complexes], and final reduction to Au(0). Subtoxic concentrations (up to 130 μM) of AuCl(4)(-) in the growth medium increase growth of R. oryzae and induce two stress response proteins while simultaneously down-regulating two other proteins. The induction increases mycelial growth, protein yield, and AuNP biosynthesis. At higher Au(III) concentrations (>130 μM), both mycelial and protein yield decrease and damages to the cellular ultrastructure are observed, likely due to the toxic effect of Au(III). Protein profile analysis also confirms the gold toxicity on R. oryzae at high concentrations. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis shows that two proteins of 45 and 42 kDa participate in gold reduction, while an 80 kDa protein serves as a capping agent in AuNP biosynthesis.

Over the last decade, advances in diagnostic systems and the introduction of new antifungal agents have significantly improved outcomes in immunocompromised patients who develop invasive aspergillosis. However, mortality rates remain relatively unchanged for less common, but highly aggressive, mold infections such as mucormycosis. Recent genome sequencing of Rhizopus oryzae revealed evidence of a whole-genome duplication event during the evolution of this pathogen. Consequently, R. oryzae has a 2- to 10-fold enrichment in gene families associated with ergosterol and cell wall biosynthesis, cell growth, iron uptake, and known fungal virulence factors compared with sequenced Aspergillus fumigatus strains. This genetic plasticity may explain the remarkable capability of this pathogen for rapid growth in hostile environments, such as the inflammatory milieu, as well as its relative resistance to multiple antifungal classes. Herein, we examine how pharmacological aspects of treating mucormycosis may differ from those of the more commonly encountered invasive aspergillosis.

Two nectarine cultivars ('Flavela' and 'Flanoba') were treated with Aloe vera gel alone, or with the addition of thymol, and then inoculated with Rhizopus stolonifer, Botrytis cinerea and Penicillium digitatum. Both treatments were effective in reducing the decay incidence caused by the 3 fungi species, although the addition of thymol did not generally improve the efficacy of Aloe vera gel on reducing the infection damage. The coatings were clearly effective in reducing the postharvest ripening process of both nectarine cultivars manifested by a delay in ethylene production and respiration rate, weight loss and softening. Interestingly, these coatings showed effectiveness on reducing decay development in inoculated fruits and thus Aloe vera could be considered as natural antifungal compound and might serve as alternative of synthetic fungicides., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The saprophytic fungus Rhizopus nigricans constitutes a serious problem when thriving on gathered crops. The identification of any compounds, especially natural ones, that inhibit fungal growth, may therefore be important. During its life cycle, Rhizopus nigricans encounters many compounds, among them the flavonoids, plant secondary metabolites that are involved in plant defense against pathogenic microorganisms. Although not being a plant pathogen, Rhizopus nigricans may interact with these compounds in the same way as plant pathogens--in response to the fungitoxic effect of flavonoids the fungi transform them into less toxic metabolites. We have studied the interaction of R. nigricans with some flavonoids. Inhibition of hyphal spreading (from 3% to 100%) was observed by 300 μM flavones, flavanones and isoflavones, irrespective of their basic structure, oxidized or reduced C-ring, and orientation of the B-ring. However, a hydrophobic A-ring was important for the toxicity. R. nigricans transformed some of the flavonoids into glucosylated products. Recognition of substrates for glucosylating enzyme(s) did not correlate with their fungitoxic effect but depended exclusively on the presence of a free -OH group in the flavonoid A-ring and of a hydrophobic B-ring. Although the fungus produced glucosyltransferase constitutively, an additional amount of the enzyme was induced by the substrate flavonoid. Moreover, effective detoxification was shown to require the presence of glucose., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The antagonistic activities of Bacillus subtilis fmbj against the Rhizopus stolonifer pathogen causing peach soft rot disease were studied in this paper. Bacillus subtilis fmbj exhibited a high antifungal effect on the mycelium growth, sportulation, and germ tube elongation of R. stolonifer with the inhibition rate of 86.5% and 97.42%, respectively, spore germination rate of 20% under antagonist strain concentration of 10⁸ CFU/mL. By using of scanning electron microscope (SEM) and transmission electron microscope (TEM), it was observed that B. subtilis fmbj strain strongly induced morphological abnormalities of R. stolonifer and destroyed structure of hypha and spore. When hypha cell wall of R. stolonifer was damaged, the organelles and cytoplasms inside cell would exude, which led to cell death., (© 2011 Institute of Food Technologists®)

Fifteen fungal strains were isolated from arsenic contaminated (range 9.45-15.63 mg kg(-1)) agricultural soils from the state of West Bengal, India. Five fungal strains were belonged to the Aspergillus and Trichoderma group each, however, remaining five were identified as the Neocosmospora, Sordaria, Rhizopus, Penicillium and sterile mycelial strain. All these fungal strains were cultivated on medium supplemented with 100, 500, 1000, 5000 and 10,000 mg l(-1) of sodium arsenate. After 30-day cultivation under laboratory conditions, radial growth of these strains was determined and compared with control. Toxicity and tolerance of these strains to arsenate were evaluated on the basis of tolerance index. Out of fifteen, only five fungal strains were found resistant and survived with tolerance index pattern as 0.956 (sterile mycelial strain)>0.311 (Rhizopus sp.)>0.306 (Neocosmospora sp.)>0.212 (Penicillium sp.)>0.189 (Aspergillus sp.) at 10,000 mg l(-1) of arsenate. The arsenic removal efficacy of ten fungal strains, tolerant to 5000 mg l(-1) arsenate, was also assayed under laboratory conditions for 21 days. All these strains were cultivated individually on mycological broth enriched with 10 mg l(-1) of arsenic. The initial and final pH of cultivating medium, fungal biomass and removal of arsenic by each fungal strain were evaluated. Fungal biomass of ten strains removed arsenic biologically from the medium which were ranged from 10.92 to 65.81% depending on fungal species. The flux of biovolatilized arsenic was determined indirectly by estimating the sum of arsenic content in fungal biomass and medium. The mean percent removal as flux of biovolatilized arsenic ranged from 3.71 to 29.86%. The most effective removal of arsenic was observed in the Trichoderma sp., sterile mycelial strain, Neocosmospora sp. and Rhizopus sp. fungal strains. These fungal strains can be effectively used for the bioremediation of arsenic-contaminated agricultural soils., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A novel voltammetric electronic tongue, smart tongue, was employed to monitor the growth of mold and to differentiate between four types of mold grown in liquid medium. Principal component analysis (PCA) was used to extract the relevant information obtained by the smart tongue. Reference growth curves were based on measurements of dry weight and pH. The growth detected by the smart tongue was basically consistent with that observed by the measurement of dry weight and pH. The optimal combinations of electrodes and frequencies for monitoring growth were as follows: for Aspergillus, both the Pt and Au electrodes at 1 Hz, 10 Hz and 100 Hz; for Penicillium, the Pt and W electrodes at 100 Hz; for Mucor, the Pt, Pd and W electrodes at the three frequency segments; for Rhizopus, the Pd, Ti and Ag electrodes at the three frequency segments. The Ag electrode at 10 Hz or 100 Hz frequency could differentiate well between the four types of mold for culturing 6 h in the liquid media. Therefore, the smart tongue has a promising future as a modern rapid analytical technology for the real time detection of the growth of mold and for the classification model of mold., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Lactic acid, an anaerobic product of glycolysis, can be theoretically produced by synthetic route; however, it is commercially produced by homo-fermentative batch mode of operations. Factors affecting its production and strategies improving it are considered while devising an optimized protocol. Although a hetero-fermentative mode of production exists, it is rarely used for commercial production. Attempts to use Rhizopus sp. for lactic acid production through either hetero-fermentative or thermophilic conditions were not economical. Since almost 70% of the cost of its production is accounted by raw materials, R & D efforts are still focused to find economically attractive agri-products to serve as sources of carbon and complex nitrogen inputs to meet fastidious nutrient needs for microbial growth and lactic acid production. Therefore, need exists for using multi-pronged strategies for higher productivity. Its present production and consumption scenario is examined. Its optically active isomers and chemical structure permit its use for the production of several industrially important chemicals, health products (probiotics), food preservatives, and bio-plastics. In addition, its salts and esters appear to have a variety of applications.

Dicarboxylic acids that are produced from renewable resources are becoming attractive building blocks for the polymers industry. In this respect, fumaric acid is very interesting. Its low aqueous solubility facilitates product recovery. To avoid excessive waste salt production during downstream processing, a low pH for fumaric acid fermentation will be beneficial. Studying the influence of pH, working volume and shaking frequency on cell cultivation helped us to identify the best conditions to obtain appropriate pellet morphologies of a wild type strain of Rhizopus oryzae. Using these pellets, the effects of pH and CO(2) addition were studied to determine the best conditions to produce fumaric acid in batch fermentations under nitrogen-limited conditions with glucose as carbon source. Decreasing either the fermentation pH below 5 or increasing the CO(2) content of the inlet air above 10% was unfavourable for the cell-specific productivity, fumaric acid yield, and fumaric acid titer. However, switching off the pH control late in the batch phase did not affect these performance parameters and allowed achieving pH of 3.6. A concentration of 20 gL(-1) of fumaric acid was obtained at pH 3.6 while the average cell mass specific productivity and fumaric acid yield were the same as at pH 5.0. Consequently, relatively modest amounts of inorganic base were required for pH control, while recovery of the acid should be relatively easy at pH 3.6., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Banagrass (Pennisetum purpureum) approximately 4 months old was hand-harvested and shredded. Half of the sample was dewatered using a screw-press, whereby the extracted juice was used for cultivating an edible fungus, Rhizopus microsporus, for aquaculture feed supplementation. The remaining biomass was divided into four separate streams: (1) wet, juiced; (2) dry, juiced; (3) wet, unjuiced; and (4) dry, unjuiced. Each stream was pretreated with dilute sulfuric acid and compared on the basis of sugar release at varying acid concentrations, temperatures, and residence times. Wet, juiced banagrass released the most soluble sugars (theoretical xylose and ∼85% glucose). Ultrasonication (20 kHz) was applied to further increase monomeric sugar release but demonstrated little improvement on total sugar yields. Fungal biomass generated from banagrass juice exhibited potential as a fungal-protein production medium producing 1.16±0.34 g biomass increase/g initial biomass., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

This study examined the potential of producing a protein-rich fungus, Rhizopus microsporus (var. oligosporus), as an aquaculture feed ingredient on vinasse generated during sugar-based ethanol fermentation. Optimization studies showed prolific fungal growth at pH 5.0 and 30 degrees C on vinasse with nutrient (nitrogen and phosphorus) supplementation. The molasses-vinasse resulted in the highest specific fungal biomass yield of 0.21 (g biomass increase/(g initial biomassxg soluble chemical oxygen demand (SCOD) removed)). Organic and inorganic matters were significantly reduced, e.g., 42.02% SCOD and 24.41% total dissolved solids. Thus, the treated effluent could be recycled as process water or used for land applications. The fungal biomass was found to have 45.55% crude protein with significantly high amino acids, arginine and threonine. The integration of innovative fungal technology with sugar-based ethanol production could provide an opportunity for producing food-grade fungal protein for animal feed application with simultaneous water reclamation., (2010 Elsevier Ltd. All rights reserved.)

Procedures for the production of endo-1,4-beta-xylanase have been developed. An active producer-Rhizopus var. microsporus BKMF-595-has been chosen, and the conditions of surface and submerged cultivation, as well as the composition of the culture medium for this strain, have been optimized to ensure maximum yield of the target enzyme. Activity of xylomicrosporin Px equaled 123 U/g, while the activity of xylomicrosporin Gx equaled 25 U/cm3. Homogeneous enzyme preparations, purified 59.44-fold and 72.6-fold, have been obtained. The dependence of endo-1,4-beta-xylanase catalytic activity on temperature and pH of the reaction medium has been studied. The enzyme has been shown to be most stable in the pH range 5.0-6.0 and to be thermostable. Amino acid composition and subunit structure of the enzyme were determined; the molecular masses of the subunits equaled 50 and 56 kDa. Carboxyl groups of glutamic and aspartic acid residues of the active center were experimentally shown to play an important role in catalysis. The potential of this enzyme for beer production has been demonstrated.

Aims: To examine the potential use of hemicellulose hydrolysate (HH) for the production of chitosan by Rhizopus oryzae and investigate the influence of contents in HH on mycelia growth and chitosan synthesis., Methods and Results: Compared to xylose medium, HH enhanced mycelia growth, chitosan content and production of R. oryzae by 10.2, 64.5 and 82.1%, respectively. During sulfuric acid hydrolysis of corn straw, sugars (glucose, galactose, etc) and inhibitors (formic acid, acetic acid and furfural) were generated. Acetic acid (2.14 g l(-1)) and formic acid (0.83 g l(-1)) were stimulative, while furfural (0.55 g l(-1)) was inhibitory. Inhibitors, at different concentrations, increased the mycelia growth and chitosan production by 24.5-37.8 and 60.1-207.1%., Conclusions: HH of corn straw is a good source for chitosan production. Inhibitors in HH, at proper concentrations, can enhance chitosan production greatly., Significance and Impact of the Study: This work for the first time reported chitosan production from HH. Chitosan production can be greatly enhanced by cheap chemicals such as inhibitors in HH.

Mucormycosis is a fungal infection of the sinuses, brain, or lungs that causes a mortality rate of at least 50% despite first-line therapy. Because angioinvasion is a hallmark of mucormycosis infections, we sought to define the endothelial cell receptor(s) for fungi of the order Mucorales (the fungi that cause mucormycosis). Furthermore, since patients with elevated available serum iron, including those with diabetic ketoacidosis (DKA), are uniquely susceptible to mucormycosis, we sought to define the role of iron and glucose in regulating the expression of such a receptor. Here, we have identified glucose-regulated protein 78 (GRP78) as what we believe to be a novel host receptor that mediates invasion and damage of human endothelial cells by Rhizopus oryzae, the most common etiologic species of Mucorales, but not Candida albicans or Aspergillus fumigatus. Elevated concentrations of glucose and iron, consistent with those seen during DKA, enhanced GRP78 expression and the resulting R. oryzae invasion and damage of endothelial cells in a receptor-dependent manner. Mice with DKA, which have enhanced susceptibility to mucormycosis, exhibited increased expression of GRP78 in sinus, lungs, and brain compared with normal mice. Finally, GRP78-specific immune serum protected mice with DKA from mucormycosis. These results suggest a unique susceptibility of patients with DKA to mucormycosis and provide a foundation for the development of new therapeutic interventions for these deadly infections.