scholarly journals Production and Optimization of Xylanase and α-Amylase from Non-Saccharomyces Yeasts (Pichia membranifaciens)

2021 ◽  
pp. 452-461
Author(s):  
Hala A. Salah ◽  
Hanan A. Temerk ◽  
Nivin A. Salah ◽  
Saeed Rafa Zara Alshehri ◽  
Jazi A. Al-Harbi ◽  
...  
Keyword(s):  
Culture Medium ◽  
Potato Starch ◽  
Pcr Amplification ◽  
Soluble Starch ◽  
Relative Activity ◽  
Rrna Gene ◽  
Pichia Membranifaciens ◽  
Ph Values ◽  
26S Rrna ◽  
Saccharomyces Yeasts

The xylanolytic and amylolytic yeasts were qualitatively determined by Cong red xylan agar and soluble starch agar plates, respectively. The most xylanase and α-amylase inducible strain (AUN-02) was selected and identified using PCR amplification of 26S rRNA gene and sequence analysis. The comparison of the alignment results and phylogenetic analysis of the sequences of the isolated yeast to published rRNA gene sequences in GenBank, confirmed the identification of the isolate as Pichia membranifaciens. Xylanase and α-amylase production by isolated P. membranifaciens were investigated at different pH values (4-8), temperature degrees (20-45°C), incubation time (1-7 days) and various substrates.A higher production of xylanase (38.8 U/mL) and a-amylase (28.7 U/mL) was obtained after 4 days of fermentation of P. membranifaciens. Higher activity of xylanase (36.83 U/mL) and a-amylase (27.7 U/mL) was obtained in the fermentation of P. membranifaciens in a culture medium adjusted to pH 7.0. The optimum temperature showed maximum xylanase and a-amylase activity (42.6 and 32.5 units/mL, respectively) was estimated at 35 °C. The xylanase and a-amylase activities of P. membranifaciens were estimated and compared for the different substrates tested. The strain revealed 100% relative activity of xylanase and a-amylase on beechwood and potato starch, respectively. The affinity of enzymes towards substrate was estimated using Km values. The Km values of xylanase and α-amylase increased in the order of pH’s 7.0, 6.0 and 4.5 (0.85, 1.6 and 3.4 mg xylan/mL and 0.22, 0.43 and 2.8 mg starch/mL, respectively). the yeast P. membranifaciensis is suitable for produce neutral xylanase and α-amylase enzymes. So, it could be used as a promising strain for production of these enzymes in industrial field.

scholarly journals Adhesion Properties, Biofilm Forming Potential, and Susceptibility to Disinfectants of Contaminant Wine Yeasts

2021 ◽  
Vol 9 (3) ◽  
pp. 654
Author(s):  
Giorgia Perpetuini ◽  
Alessio Pio Rossetti ◽  
Noemi Battistelli ◽  
Giuseppe Arfelli ◽  
Rosanna Tofalo
Keyword(s):  
Stainless Steel ◽  
Restriction Analysis ◽  
Rrna Gene ◽  
Pichia Kudriavzevii ◽  
Adhesion Properties ◽  
Pichia Membranifaciens ◽  
Potassium Metabisulphite ◽  
D2 Domain ◽  
Pichia Manshurica ◽  
26S Rrna

In this study, yeasts isolated from filter membranes used for the quality control of bottled wines were identified and tested for their resistance to some cleaning agents and potassium metabisulphite, adhesion to polystyrene and stainless-steel surfaces, and formation of a thin round biofilm, referred to as a MAT. A total of 40 strains were identified by rRNA internal transcribed spacer (ITS) restriction analysis and sequence analysis of D1/D2 domain of 26S rRNA gene. Strains belong to Pichia manshurica (12), Pichia kudriavzevii (9), Pichia membranifaciens (1), Candida sojae (6), Candida parapsilosis (3), Candida sonorensis (1), Lodderomyces elongisporus (2), Sporopachydermia lactativora (3), and Clavispora lusitaniae (3) species. Regarding the adhesion properties, differences were observed among species. Yeasts preferred planktonic state when tested on polystyrene plates. On stainless-steel supports, adhered cells reached values of about 6 log CFU/mL. MAT structures were formed only by yeasts belonging to the Pichia genus. Yeast species showed different resistance to sanitizers, with peracetic acid being the most effective and active at low concentrations, with minimum inhibitory concentration (MIC) values ranging from 0.08% (v/v) to 1% (v/v). C. parapsilosis was the most sensible species. Data could be exploited to develop sustainable strategies to reduce wine contamination and establish tailored sanitizing procedures.

scholarly journals Molecular screening for Candida orthopsilosis and Candidametapsilosis among Danish Candida parapsilosis group blood cultureisolates: proposal of a new RFLP profile for differentiation

2010 ◽  
Vol 59 (4) ◽  
pp. 414-420 ◽  
Author(s):  
Hossein Mirhendi ◽  
Brita Bruun ◽  
Henrik Carl Schønheyder ◽  
Jens Jørgen Christensen ◽  
Kurt Fuursted ◽  
...  
Keyword(s):  
Candida Parapsilosis ◽  
Profile Analysis ◽  
Secondary Alcohol ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Rflp Profile ◽  
Pcr Rflp ◽  
Invasive Infections ◽  
Candida Orthopsilosis ◽  
26S Rrna

Candida orthopsilosis and Candida metapsilosis are recentlydescribed species phenotypically indistinguishable from Candida parapsilosis. We evaluated phenotyping and molecular methods for the detection ofthese species among 79 unique blood culture isolates of the C. parapsilosis group obtained during the years 2004–2008. The isolates were screenedby PCR amplification of the secondary alcohol dehydrogenase-encoding gene (SADH) followed by digestion with the restriction enzyme BanI, using C. parapsilosis ATCC 22019, C. orthopsilosisATCC 96139 and C. metapsilosis ATCC 96144 as controls. Isolates withRFLP patterns distinct from C. parapsilosis were characterized bysequence analysis of the ITS1–ITS2, 26S rRNA (D1/D2) and SADH regions. Restriction patterns for the 3 species with each of 610restriction enzymes were predicted in silico using 12 available sequences.By PCR-RFLP of the SADH gene alone, four isolates (5.1 %)had a pattern identical to the C. orthopsilosis reference strain.Sequence analysis of SADH and ITS (internal transcribed spacer)regions identified two of these isolates as C. metapsilosis. Theseresults were confirmed by creating a phylogenetic tree based on concatenatedsequences of SADH, ITS and 26S rRNA gene sequence regions. Optimaldifferentiation between C. parapsilosis, C. metapsilosisand C. orthopsilosis was predicted using digestion with NlaIII,producing discriminatory band sizes of: 131 and 505 bp; 74, 288 and 348 bp;and 131, 217 and 288 bp, respectively. This was confirmed using the referencestrains and 79 clinical isolates. In conclusion, reliable discrimination wasobtained by PCR-RFLP profile analysis of the SADH gene after digestionwith NlaIII but not with BanI. C. metapsilosisand C. orthopsilosis are involved in a small but significant numberof invasive infections in Denmark.

scholarly journals Cryptococcus thermophilus sp. nov., isolated from cassava sourdough

2012 ◽  
Vol 62 (Pt_7) ◽  
pp. 1715-1720 ◽  
Author(s):  
Stephanie A. Vogelmann ◽  
Sandra Chaves ◽  
Christian Hertel
Keyword(s):  
Sodium Nitrite ◽  
Type Strain ◽  
Novel Species ◽  
Soluble Starch ◽  
Rrna Gene ◽  
The Novel ◽  
Cassava Flour ◽  
D2 Domain ◽  
Anamorphic Yeast ◽  
26S Rrna

A novel anamorphic yeast, strain LTH 6662T, was isolated from cassava sourdough. The isolate supposedly originated from cassava flour or was a contaminant thereof. Sequencing of the D1/D2 domain of the 26S rRNA gene indicated that strain LTH 6662T represents a novel species. Its closest relatives were members of the Cryptococcus humicola complex. The novel strain had several physiological characteristics that differed from those of related species: the ability to assimilate raffinose and cadaverine; the inability to assimilate soluble starch, xylitol, galactitol, butane-2,3-diol, sodium nitrite and lysine; the ability to grow without vitamins and at 42 °C; and the inability to produce starch-like substances. Its major ubiquinone was Q-10. In addition, buds were formed on small neck-like structures. In liquid medium, green or blue fluorescent substances were produced. The name Cryptococcus thermophilus sp. nov. is proposed, with LTH 6662T ( = DSM 19443T = CBS 10687T) as the type strain.

STUDIES ON THE PHYSIOLOGY OF ELSINOE VENETA

1953 ◽  
Vol 31 (4) ◽  
pp. 411-422 ◽  
Author(s):  
W. G. Kemp
Keyword(s):  
Potato Starch ◽  
Nitrogen Compound ◽  
Maximum Growth ◽  
Sole Source ◽  
Soluble Starch ◽  
Continuous Darkness ◽  
Ph Values ◽  
Wide Range ◽  
Initial Ph ◽  
Fungus Growth

A study was made of the effect of certain nutritional and environmental factors on the growth and pigmentation of the mycelium, the sporulation, and the germination of the conidia of a "convoluted" isolate of Elsinoe veneta (Burkh.) Jenkins, the fungus responsible for the anthracnose disease of raspberries. This isolate utilized with varying degrees of efficiency various mono-, oligo-, and polysaccharides as well as certain organic alcohols as its sole source of carbon for growth. Either a nitrate, ammonium, amino, or imidazole nitrogen compound supported the fungus in culture. Maximum growth of the mycelium occurred in the presence of soluble starch and asparagine, whereas optimum sporulation of the conidia was obtained on media containing potato starch and sodium nitrate. In general, the production of conidia was markedly reduced on media favorable for excessive vegetative growth. Both a decrease in the volume of the medium and in the concentration of either a specific nutrient or of total nutrients adversely influenced the production of mycelium. Temperatures above 30 °C. and below 21 °C. decreased the percentage germination of conidia and restricted the growth of the fungus. Growth and sporulation occurred over a wide range of pH values. The optimum initial pH for both growth and sporulation was 4.0. E. veneta produced conidia and grew as well in continuous darkness as in alternate diffuse light and darkness. Young potted raspberry plants growing in the greenhouse, when inoculated with conidia produced in culture, developed typical anthracnose lesions on the canes.

scholarly journals New Chitosan-Degrading Strains That Produce Chitosanases Similar to ChoA of Mitsuaria chitosanitabida

2005 ◽  
Vol 71 (9) ◽  
pp. 5138-5144 ◽  
Author(s):  
ChoongSoo Yun ◽  
Daiki Amakata ◽  
Yasuhiro Matsuo ◽  
Hideyuki Matsuda ◽  
Makoto Kawamukai
Keyword(s):  
Southern Hybridization ◽  
Glycosyl Hydrolase ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
16S Rrna Gene Sequences ◽  
Glycosyl Hydrolase Family ◽  
Broad Variety ◽  
Hydrolase Family ◽  
The 16S Rrna Gene ◽  
Sequence Similarities

ABSTRACT The betaproteobacterium Mitsuaria chitosanitabida (formerly Matsuebacter chitosanotabidus) 3001 produces a chitosanase (ChoA) that is classified in glycosyl hydrolase family 80. While many chitosanase genes have been isolated from various bacteria to date, they show limited homology to the M. chitosanitabida 3001 chitosanase gene (choA). To investigate the phylogenetic distribution of chitosanases analogous to ChoA in nature, we identified 67 chitosan-degrading strains by screening and investigated their physiological and biological characteristics. We then searched for similarities to ChoA by Western blotting and Southern hybridization and selected 11 strains whose chitosanases showed the most similarity to ChoA. PCR amplification and sequencing of the chitosanase genes from these strains revealed high deduced amino acid sequence similarities to ChoA ranging from 77% to 99%. Analysis of the 16S rRNA gene sequences of the 11 selected strains indicated that they are widely distributed in the β and γ subclasses of Proteobacteria and the Flavobacterium group. These observations suggest that the ChoA-like chitosanases that belong to family 80 occur widely in a broad variety of bacteria.

scholarly journals In Vitro Incorporation of Helicobacter pylori into Candida albicans Caused by Acidic pH Stress

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 489 ◽  
Author(s):  
Kimberly Sánchez-Alonzo ◽  
Cristian Parra-Sepúlveda ◽  
Samuel Vega ◽  
Humberto Bernasconi ◽  
Víctor L. Campos ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Candida Albicans ◽  
Pcr Amplification ◽  
Growth Curves ◽  
Acidic Ph ◽  
Ph Values ◽  
16S Gene ◽  
Wide Range ◽  
H Pylori

Yeasts can adapt to a wide range of pH fluctuations (2 to 10), while Helicobacter pylori, a facultative intracellular bacterium, can adapt to a range from pH 6 to 8. This work analyzed if H. pylori J99 can protect itself from acidic pH by entering into Candida albicans ATCC 90028. Growth curves were determined for H. pylori and C. albicans at pH 3, 4, and 7. Both microorganisms were co-incubated at the same pH values, and the presence of intra-yeast bacteria was evaluated. Intra-yeast bacteria-like bodies were detected using wet mounting, and intra-yeast binding of anti-H. pylori antibodies was detected using immunofluorescence. The presence of the H. pylori rDNA 16S gene in total DNA from yeasts was demonstrated after PCR amplification. H. pylori showed larger death percentages at pH 3 and 4 than at pH 7. On the contrary, the viability of the yeast was not affected by any of the pHs evaluated. H. pylori entered into C. albicans at all the pH values assayed but to a greater extent at unfavorable pH values (pH 3 or 4, p = 0.014 and p = 0.001, respectively). In conclusion, it is possible to suggest that H. pylori can shelter itself within C. albicans under unfavorable pH conditions.

scholarly journals Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

2003 ◽  
Vol 69 (12) ◽  
pp. 7430-7434 ◽  
Author(s):  
Trevor G. Phister ◽  
David A. Mills
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Primers ◽  
Rrna Gene ◽  
Dekkera Bruxellensis ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Spoilage Yeast ◽  
Pcr Method ◽  
26S Rrna

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.

scholarly journals Water quality and antifungal susceptibility of opportunistic yeast pathogens from rivers

2016 ◽  
Vol 75 (6) ◽  
pp. 1319-1331 ◽  
Author(s):  
M. E. Monapathi ◽  
C. C. Bezuidenhout ◽  
O. H. J. Rhode
Keyword(s):  
Water Quality ◽  
Antifungal Susceptibility ◽  
Principal Component ◽  
Water Sources ◽  
Health Concern ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Livestock Farming ◽  
Physico Chemical ◽  
26S Rrna

Yeasts from water sources have been associated with diseases ranging from superficial mucosal infections to life threatening diseases. The aim of this study was to determine the water quality as well as diversity and antifungal susceptibility of yeasts from two rivers. Yeast levels and physico-chemical parameter data were analyzed by principal component analysis to determine correlations between physico-chemical data and yeast levels. Yeast morphotypes were identified by biochemical tests and 26S rRNA gene sequencing. Disk diffusion antifungal susceptibility tests were conducted. Physico-chemical parameters of the water were within target water quality range (TWQR) for livestock farming. For irrigational use, total dissolved solids and nitrates were not within the TWQR. Yeast levels ranged between 27 ± 10 and 2,573 ± 306 cfu/L. Only non-pigmented, ascomycetous yeasts were isolated. Saccharomyces cerevisiae and Candida glabrata were most frequently isolated. Several other opportunistic pathogens were also isolated. A large number of isolates were resistant to azoles, especially fluconazole, but also to other antifungal classes. Candida species were resistant to almost all the antifungal classes. These water sources are used for recreation and religious as well as for watering livestock and irrigation. Of particular concern is the direct contact of individuals with opportunistic yeast, especially the immune-compromised. Resistance of these yeast species to antifungal agents is a further health concern.

Two main clusters within Trypanosoma cruzi zymodeme 3 are defined by distinct regions of the ribosomal RNA cistron

Parasitology ◽  
2002 ◽  
Vol 124 (2) ◽  
pp. 177-184 ◽  
Author(s):  
M. B. A. MENDONÇA ◽  
N. S. NEHME ◽  
S. S. SANTOS ◽  
E. CUPOLILLO ◽  
N. VARGAS ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Ribosomal Rna ◽  
Southern Blotting ◽  
Pcr Amplification ◽  
Rrna Gene ◽  
Pcr Products ◽  
Phylogenetic Lineages ◽  
Definition Of ◽  
Ribosomal Cistron ◽  
Panstrongylus Geniculatus

Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3–Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Sα rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5·8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Sα rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates.

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