Citeromyces Matritensis Research Articles

Fungal dynamic changes in naturally fermented 'Kyoho' grape juice.

The 'Kyoho' grape (Vitaceae, Plantae) has large ears, plenty of flesh, and rich nutrition and is planted across a large area in China. There are few reports on this variety in winemaking, especially on the dynamic changes of fungi in the wine fermentation broth. In this study, we used the 'Kyoho' grapes as raw materials and adopted a high throughput to analyze dynamic changes in fungal species composition of the natural fermentation broth at four time points: day 1 (D1P), day 3 (D3P), day 5 (D5P), and day 15 (D15P). Changes in fungal metabolic pathways and dominant yeasts were also analyzed. A total of 78 families, 110 genera, and 137 species were detected, in the natural fermentation broth samples. Forty-nine families, 60 genera, and 72 species were found in the control check (CK). A total of 66 differential metabolic pathways were enriched; of those, 41 were up-regulated compared to CK, such as CDP-diacylglycerol biosynthesis I (PWY 5667), chitin degradation to ethanol (PWY 7118), and the super pathway of phosphatidate biosynthesis (PWY 7411). Changes in fungal metabolic pathways were in line with the dynamic changes of dominant yeast species in the whole process of fermentation. Pichia kluyveri, P. membranifaciens, and Citeromyces matritensis are the dominant species in the later stages of natural fermentation. These yeast species may play vital roles in the 'Kyoho' wine industry in the future.

Insights Into the Cultivable Microbial Ecology of "Manna" Ash Products Extracted From Fraxinus angustifolia (Oleaceae) Trees in Sicily, Italy.

Microbial communities characterizing a specific food matrix, generally, strongly contribute to both its composition, and properties for food applications. To our knowledge, this is the first study to investigate the cultivable microbial ecology of Sicilian “Manna” ash products in order to acquire new information on the hygienic quality, shelf-life and potential application of this traditional food. To this purpose, several manna samples belonging to different commercial categories were collected and subjected to the analysis of bacteria, yeasts, and filamentous fungi. Furthermore, an investigation of the sugar content and physicochemical parameters was performed. The results of our study followed the trend generally reported for other sugary foods. Conversely, as regards microbiological analyses, in the present study, the presence of microorganisms at high levels confirmed their survival in stressing conditions characterizing this food matrix in a viable and cultivable form. Most species were osmophilic, endophytic bacteria, antagonistic of fungi pathogen of plants. Yeasts were the most abundant microbial populations and a total of six species were identified: Candida aaseri, Candida lactis-condensi, Citeromyces matritensis, Lachancea thermotolerans, Saccharomyces cerevisiae, and Zygosaccharomyces bailii. Filamentous fungi included five genera, which were considered common contaminants of honey and of other foods due to their xerophilic characteristics. Interestingly, our results suggest that the strains of L. thermotolerans isolated in this study might be evaluated for their potential to act as starters either singly or in multi-combination for food applications.

New epiphytic yeasts able to reduce grey mold disease on apples

Botrytis cinerea, the causal agent of grey mould, is a predominant agent causing extensive postharvest and quality losses of apples in Tunisia and worldwide. Efforts to manage this disease have met with limited success. For this reason, the use of microorganism preparations to control fungal diseases as an alternative to fungicides became an urgent need. From a total of 60 epiphytic yeasts, 10 were assessed in vitro against B. cinerea and selected isolates showing antagonism were evaluated for their ability to suppress the grey mould in vivo. On Petri plates, the most promising strains (three strains of Aureobasidium pullulans, one Cryptococcus flavescens, and one Citeromyces matritensis) showed a zone of inhibition against the pathogen fungus not exceeding 10 mm. In vivo, these isolates showed a remarkable antifungal activity since they significantly reduced disease severity on apples from 63% to 95% compared to the control. In conclusion, the work has demonstrated that the three strains, L7 of Aureobasidium pullulans, L2 of Citeromyces matritensis, and L10 of Cryptococcus flavescens, were highly effective and can be used as potential biocontrol agents in controlling the post-harvest decay of apples caused by B. cinerea.

Citeromyces matritensis M37 is a salt-tolerant yeast that produces ethanol from salted algae.

Algae are referred to as a third-generation biomass for ethanol production. However, salinity treatment is a problem that needs to be solved, because algal hydrolysates often contain high salt. Here, we isolated the salt-tolerant ethanol-producing yeast Citeromyces matritensis M37 from the east coast of Miura Peninsula in Japan. This yeast grew under osmotic stress conditions (20% NaCl or 60% glucose). It produced 6.55 g/L ethanol from YPD medium containing 15% NaCl after 48 h, and the ethanol accumulation was observed even at 20% NaCl. Using salted Undaria pinnatifida (wakame), we obtained 6.33 g/L glucose from approx. 150 g/L of the salted wakame powder with acidic and heat pretreatment followed by enzymatic saccharification, and the ethanol production reached 2.58 g/L for C. matritensis M37. The ethanol concentration was 1.4 times higher compared with that using the salt-tolerant ethanol-producing yeast Zygosaccharomyces rouxii S11.

Biotechnological activities from yeasts isolated from olive oil mills

The yeast biota associated to three eastern Spain oil mills has been evaluated. Each environment has associated a characteristic number and variability of yeasts. The D1/D2 rDNA sequencing protocol has been proved to be a useful method to identify 216 yeasts from oleic environments, showing us Candida (several species), Saccharomyces paradoxus, Citeromyces matritensis and Cryptococcus sp. as the most representative genera. Moreover, a further screening of biochemical activities has provided us with a collection of isolates able to metabolize different substrates. In particular, protease, β-glucosidase, pectinase, polygalacturonase, xylanase, lipase, esterase and catalase activities were investigated. The majority of the strains showed moderate or high β-glucosidase, lipase and pectinase activities, while protease and polygalacturonase activities are less frequent. In a second step, nutritional competition assays were carried out to investigate the antagonic effect of yeasts associated to olive oil mills with Aspergillus fungi. Yeasts can be an effective biocontrol tool as they can colonize olives and compete for space and nutrients with pathogenic fungi, avoiding the use of chemical fungicides.

Citeromyces nyonsensis sp. nov., a novel yeast species isolated from black olive brine

A yeast strain was isolated from olive brines in a fermented black olive and olive oil manufacturing plant in the town of Nyons (France). On the basis of domains 1 and 2 (D1/D2) large subunit (LSU) rRNA gene and internal transcribed spacer (ITS) region sequence analyses, the strain CLIB 1303(T) was found to be closely related, but clearly distinct, from the three existing species of the genus Citeromyces: Citeromyces matritensis, Citeromyces siamensis and Citeromyces haiwaiiensis. Strain CLIB 1303(T) exhibited 6 bp, 7 bp and 12 bp divergences in the D1/D2 LSU rRNA gene with C. siamensis, C. matritensis and C. hawaiiensis, respectively. ITS region divergence amounted to more than 8 %, 4 % and 4.5 % with C. siamensis, C. matritensis and C. hawaiiensis, respectively, in addition to several indels. Like C. matritensis and C. siamensis strains, strain CLIB 1303(T) was shown to be halotolerant and osmotolerant. Phenotypically, strain CLIB 1303(T) can be distinguished from other species of the genus Citeromyces by its inability to assimilate trehalose. The strain CLIB 1303(T) (= CBS 12700(T)) was assigned to a novel species, Citeromyces nyonsensis sp. nov.

Phenoloxidase-producing halotolerant fungi from olive brine wastewater

The main aim of this study was to assess the ability of producing extracellular phenoloxidases (EPO) of fungi isolated from olive brine wastewater (OBW), the effluent from the debittering process of table olives. Five out of twenty isolates displayed EPO-producing ability as assessed by selective agar plate assays and were all halotolerant. Among them, Citeromyces matritensis (syn. Candida globosa) and Aspergillus fumigatus concomitantly produced laccase and Mn-peroxidase (MnP) activities. Candida boidinii and Candida bombi, instead, only produced laccase and Candida diddensiae only released MnP. Both C. matritensis and A. fumigatus, grown in shaken cultures on OBW, maintained their EPO-producing ability and removed phenols by 52.3 and 82.3%, respectively, after 10d incubation in the non-supplemented effluent. These results might suggest the possible use of these strains in the treatment of other saline phenol-rich effluents, such as pickling and tannery wastewater.

Microbial quality and yeast population dynamics in cracked green table olives’ fermentations

Cracked green table olives, from the Manzanilla variety, are a fermented food produced and consumed in Portugal. The objective of the present work was to study the microbiological characteristics and yeast population evolution during the fermentation of cracked green olives. The predominant microorganisms were yeasts while lactic acid bacteria were not detected and a clear decrease of the mould population was observed. At the end of the fermentations, no viable counts of Enterobacteriaceae were found. Yeast isolates were identified by the 5.8S rRNA-ITS region restriction analysis and by sequencing the D1/D2 region of the 26S rRNA gene. During the initial phases of the fermentations a great diversity of yeasts was observed. However, as the processes evolved the biodiversity decreased with the fermentative yeasts Citeromyces matritensis, Zygotorulaspora mrakii and Saccharomyces cerevisiae becoming the dominant species. The presence of these fermentative yeasts at the end of the production process is associated to a risk of spoilage. The results obtained represent a first attempt towards the comprehension of the microbiota of this type of “Natural olives” that constitute an important component of the Mediterranean diet.

Proteolytic activities of bacteria, yeasts and filamentous fungi isolated from coffee fruit (Coffea arabica L.)

One hundred forty-four microorganisms previously isolated from coffee fruit (Coffea arabica) were grown on casein agar to evaluate their proteolytic activities. Fifty percent of filamentous fungi, 52.5% of bacteria and 2.6% of yeasts were able to secrete proteases. Positive isolates were further examined in liquid culture for their protease activities by hydrolysis of casein at different pH values (5.0, 7.0 and 9.0) at 30 o C. Bacillus megaterium, B. subtilis, Enterobacter agglomerans, Kurthia sp, Pseudomonas paucimobilis and Tatumella ptyseos demonstrated the highest proteolytic activities at pH 9.0. One yeast isolate, Citeromyces matritensis, had a proteolytic activity of 2.40 U at pH 5.0. Aspergillus dimorphicus, A. ochraceus, Fusarium moniliforme, F. solani, Penicillium fellutanum and P. waksmanii showed the highest activities. Of the bacterial isolates, the highest enzyme activities were observed in B. subtilis 333 (27.1 U), Tatumella ptyseos (27.0 U) and B. megaterium 817 (26.2 U). Of the filamentous fungi, Aspergillus ochraceus (48.7 U), Fusarium moniliforme 221 (37.5 U) and F. solani 359 (37.4 U) had the highest activities at pH 9.0.

Metallization of bacteria cells

Bacteria cells with different standard shapes are well suited for use as templates for the fabrication of magnetic and electrically conductive microstructures. In this paper, metallization of bacteria cells is demonstrated by an electroless deposition technique of nickel-phosphorus initiated by colloid palladium-tin catalyst on the surfaces of Citeromyces matritensis and Bacillus cereus. The activated and metallized bacteria cells have been characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction analysis (XRD). Results showed that both Citeromyces matritensis and Bacillus cereus had no deformation in shape after metallization; the metallized films deposited on the surfaces of bacteria cells are homogeneous in thickness and noncrystalline in phase structure. The kinetics of colloid palladium-tin solution and electroless plating on bacteria cells is discussed.

Yeast populations on Spanish fermented sausages

Yeast populations on 24 lots of Spanish fermented sausages, made by four factories (F1, F2 and F4, artisanal; F3, industrial) were investigated throughout manufacture and the influence of different variables evaluated. In addition, 41 yeast strains were identified at the species level using two miniaturised systems: ATB32C (API System) and Vitek Yeast Biochemical Card (Vitek YBC). Levels of yeasts found in the sausage mixture (mean counts around 4 log units/g) were similar to those described by other authors. In sausages from factories F1 and F2, a further increase was noted, reaching 5.5 log units/g after fermentation. Counts subsequently decreased to 3.6 and 5 log units/g, respectively. In sausages from factories F3 and F4, decreasing counts were observed from the beginning, particularly in sausages from F3, where yeasts were almost absent in the finished product. Type of manufacture and sausage diameter, were the variables most influencing yeast counts. Debaryomyces hansenii (teleomorph of C. famata) was the dominant species, being found at all stages of manufacture. Trichosporon ovoides (formerly T. beigelii), Yarrowia lipolytica (perfect form of C. lipolytica), C. intermedia/curvata, C. parapsilosis, C. zeylanoides and Citeromyces matritensis (teleomorph of C. globosa) were also present. Direct identification was possible only with 50% of the total of strains investigated, although a higher number of strains was identified using the API than the Vitek YBC system.

A phylogenetic study of ubiquinone Q-8 species of the genera Candida, Pichia, and Citeromyces based on 18S ribosomal DNA sequence divergence.

To clarify phylogenetic relationships among species of the anamorphic ascomycetous genus Candida with ubiquinone Q-8, we determined complete sequences of 18S ribosomal RNA genes (18S rDNAs) from the type strains of 20 species of the genus Candida and 7 of the teleomorphic ascomycetous genera Pichia and Citeromyces, which have Q-8 as the major ubiquinone. Q-8-forming Candida species were divided into six clusters and were phylogenetically distant from a group of Candida species that included the type species of the genus. One Q-8-forming species from each of the genera Pichia, Citeromyces, or Clavispora was included in five of six clusters. Cluster 1 comprised C. ishiwadae, C. ernobii, C. karawaiewii, C. anatomiae, C. populi, and Pichia holstii. Cluster 2 comprised C. globosa and its teleomorph, Citeromyces matritensis. Cluster 3 comprised C. molischiana and Pichia capsulata. Cluster 4 comprised C. silvanorum, C. sequanensis, C. fennica, C. entomophila, C. homilentoma, C. rhagii, C. gotoi, and Pichia burtonii. Cluster 5 comprised C. fructus, C. musae, and C. lusitaniae (anamorph of Clavispora lusitaniae). Cluster 6 comprised C. stellata, C. lactiscondensi, C. galacta, and C. incommunis and was a heterogeneous group with large interspecific divergence. Pichia pastoris was quite divergent and phylogenetically distant from other Pichia species examined. Pichia methanolica and its synonym, P. cellobiosa, which have both Q-7 and Q-8 as major ubiquinones, were closely associated with Q-7-forming Williopsis salicorniae. Based on this comparative analysis of 18S rDNA sequences, it is evident that Q-8 Candida species and Q-8 Pichia species are polyphyletic.

Production of antibacterial compounds by phylloplane-inhabiting yeasts and yeastlike fungi.

The production of antibacterial compounds by yeasts and yeastlike fungi isolated from the phylloplane is reported. Aureobasidium pullulans, Citeromyces matritensis, Cryptococcus laurentii, Rhodotorula glutinis, and Sporobolomyces roseus produced antibacterial compounds inhibitory to both Pseudomonas fluorescens and Staphylococcus aureus in an overlay bioassay. In contrast, isolates of Candida albicans, Filobasidium uniguttulatum, Saccharomyces cerevisiae, Torulaspora delbruckii, Tremella foliacea, Trichosporon beigelii, and Trichosporon dulcitum obtained from soil or from culture collections did not produce inhibitory compounds when screened by the same procedure. The production of antibacterial compounds was examined in more detail, using several isolates of A. pullulans distinguished by cluster analysis on the basis of biochemical and physiological tests. They were found to produce a range of antibacterial compounds with different activities. Two distinct antibiotics were produced by an isolate of A. pullulans in liquid culture during both the logarithmic and the stationary phases of growth.

Linear DNA plasmids of Pichia inositovora are associated with a novel killer toxin activity

Pichia inositovora, strain NRRL Y-18709, which contains three linear double-stranded DNA plasmids, pPinl-1, pPinl-2 and pPinl-3, was cured of these plasmids both by growing the strain in the presence of 50 micrograms/ml bisbenzimide, and by exposure to ultraviolet light. Both cured and uncured strains were tested for growth on a variety of carbon sources. No differences in growth response were detected, indicating no discernible involvement of the linear plasmids in the catabolism of these compounds. Culture supernatants of Pichia inositovora were shown to contain a substance larger than 100 kDa that is toxic to Saccharomyces cerevisiae, strain GS 1688. Toxin activity was optimal in YEPD assay plates containing 50 mM citrate buffer with a pH between 3.4 and 4.2. Culture supernatants from P. inositovora were also weakly active against Cephaloascus albidus, strain NRRL Y-18710, and Citeromyces matritensis, strain NRRL Y-18711. Concentrated supernatants from cured P. inositovora strains did not exhibit these activities, consistent with the hypothesis that this toxic activity is linear plasmid-encoded. Unlike the well-known Kluyveromyces lactis system, or the newly identified P. acaciae system, P. inositovora strains cured of their linear plasmids do not become detectably sensitive to toxin produced by the wild-type strain suggesting a nonplasmid-encoded immunity function.

Structural study of phosphomannan-protein complex of Citeromyces matritensis containing β-1,2 linkage : Application of partial acid degradation and acetolysis techniques under mild conditions

The phosphomannan-protein complex of Citeromyces matritensis IFO 0651 strain was investigated for its chemical structure by a sequential degradation procedure, partial acid degradation followed by acetolysis under mild conditions. Upon treatment with 10 m m HCl at 100 °C for 1 h, this complex released mannotriose and mannotetraose consisting solely of 1,2-linked β- d-mannopyranosyl residues, ca. 20% on weight basis of the parent complex. The acid-degraded complex was then subjected to acetolysis using an acetolysis medium of low sulfuric acid concentration, a 100:100:1 ( v/v) mixture of acetic anhydride, acetic acid, and sulfuric acid at 40 °C for 36 h. A phosphate-containing mannooligosaccharide fraction eluted in the void-volume region of a Bio-Gel P-2 column was found to consist of Man pβ1→2Man pβ1→ 2Man pα1→2Man to which 1 mol of phosphate group was attached, while a manno-oligosaccharide fraction eluted in the diffusable region was a mixture of Man pβ1→2Man pβ1→2Man pβ1→2Man pα1→2Man, Man pβ1 →2Man pβ1→2Man pα1→2Man, Man pβ1→2Man pα1→2Man, Man pα1→2Man, and mannose in the molar ratio of 0.08:0.33:0.19:0.32:1.00. Therefore, the structural analysis of the polysaccharide moiety of a β-1,2 linkagecontaining phosphomannan-protein complex of fungal origin can be achieved by means of a sequential degradation procedure, partial acid degradation followed by acetolysis under mild conditions.

DISTRIBUTION OF UREASE AND EXTRACELLULAR DNASE IN YEAST SPECIES

Urease and extracellular DNase activities were tested in 292 strains belonging to 125 species of 33 genera. Basidiomycetous yeasts and related species had both urease and extracellular DNase. Species belonging to the genera Rhodotorula and Cryptococcus, and four species of Candida (Candida curvata, Candida humicola, Candida bogoriensis, and Candida diffluens) which are considered to be closely related to basidiomycetous yeasts, showed both activities. In general, ascosporogenous yeasts and their imperfect forms did not have either urease or extracellular DNase. Hansenula bimundalis and Saccharomycopsis lipolytica showed both enzymes, but the activities were weaker than those of basidiomycetous yeasts. Schizosaccharomyces pombe, Schizosaccharomyces octosporus, Citeromyces matritensis, Lipomyces starkeyi, and Nadsonia elongata had urease but not extracellular DNase. Saccharomycopsis fibuligera, Hansenula jadinii, Hansenula minuta, Pichia pinus, Pichia rhodanensis, and Candida pseudotropicalis showed extracellular DNase but not urease. Taxonomic significance of urease and extracellular DNase is discussed.

Enzymic degradation of yeast cell-wall mannans and Galactomannans to polymeric fragments containing α-(1→6)-linked D-mannopyranose residues

The cell-wall D-mannans of Candida parapsilosis, Endomycopsis fibuliger, Saccharomyces rouxii, Torulopsis apicola (Hajsig strain), and Torulopsis bombi were degraded with an exo α- D-mannosidase from Arthrobacter GJM-1 to their α-(1→6)-linked D-mannopyranose main-chains, as demonstrated by p.m.r. spectroscopy. D-galacto- D-mannans from Candida lipolytica, Torulopsis gropengiesseri, Torulopsis lactis-condensi, Torulopsis magnoliae, and Trichosporon fermentans could be degraded to polysaccharides containing mainly 6- O-linked α- D-mannopyranosyl residues following preferential removal of their enzyme-resistant, D-galactopyranosyl non-reducing end-units with acid. The D-mannans of Saccharomyces lodderi, Citeromyces matritensis, and Pichia pastoris could also be enzymically degraded to polysaccharides containing predominantly α-(1→6)-linked D-mannopyranosyl residues after hydrolysis of most of the β- D-linked residues in their side chains with acid. The exo α- D-mannosidase, as would be expected, produced β- D-mannose on splitting of an α-(1→2)-linked D-mannopyranose tetramer. It is, however, very selective in its action since it did not cleave α- D-Man p-(1→2)-β- D-Man p-(1→2)-β- D-Man p-(1→2)-β- D-Man p-(1→2)- D-Man. Apparently a D-mannopyranose non-reducing end-unit and two consecutive α- D-mannopyranose residues are required by the enzyme for cleavage of a substrate to take place.

Systematics of the genera Hansenula and Pichia: proton magnetic resonance spectra of their mannans as an aid in classification.

The proton magnetic resonance (p.m.r.) spectra of the mannans extracted from a number of species of Hansenula and Pichia were determined, and the species in each genus grouped according to similarities in the p.m.r. spectra. Some of the groups of Pichia species formed mannans having similar spectra to those produced by some species of Hansenula. Several Candida species produced mannans having spectra similar to those of Pichia species. Pichia vanrijii formed a mannan whose spectrum resembled the spectra of the mannans formed by Schwanniomyces species. Pachysolen tannophilus formed a mannan with a spectrum resembling those of Hansenula holstii and Hansenula capsulata. Citeromyces matritensis and its asporogenous form Torulopsis globosa formed mannans unlike those of any other yeast studied.