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Microbial Biotechnology Mar 2017In this work, we present the development and characterization of a strain of Pichia kudriavzevii (TY1322), with highly improved phytate-degrading capacity. The mutant...
In this work, we present the development and characterization of a strain of Pichia kudriavzevii (TY1322), with highly improved phytate-degrading capacity. The mutant strain TY1322 shows a biomass-specific phytate degradation of 1.26 mmol g h after 8 h of cultivation in a high-phosphate medium, which is about 8 times higher compared with the wild-type strain. Strain TY1322 was able to grow at low pH (pH 2), at high temperature (46°C) and in the presence of ox bile (2% w/v), indicating this strain's ability to survive passage through the gastrointestinal tract. The purified phytase showed two pH optima, at pH 3.5 and 5.5, and one temperature optimum at 55°C. The lower pH optimum of 3.5 matches the reported pH of the pig stomach, meaning that TY1322 and/or its phytase is highly suitable for use in feed production. Furthermore, P. kudriavzevii TY1322 tolerates ethanol up to 6% (v/v) and shows high osmotic stress tolerance. Owing to the phenotypic characteristics and non-genetically modified organisms nature of TY1322, this strain show great potential for future uses in (i) cereal fermentations for increased mineral bioavailability, and (ii) feed production to increase the phosphate bioavailability for monogastric animals to reduce the need for artificial phosphate fortification.
Topics: 6-Phytase; Bile; Culture Media; Drug Tolerance; Enzyme Stability; Ethanol; Gene Expression Regulation, Fungal; Hydrogen-Ion Concentration; Industrial Microbiology; Mutation; Phosphates; Pichia; Temperature
PubMed: 27790831
DOI: 10.1111/1751-7915.12427 -
Probiotics and Antimicrobial Proteins Jun 2023Potential probiotic yeast strains isolated from fermented food need to meet safe and beneficial conditions for the host's health. The Pichia kudriavzevii YGM091 strain...
Potential probiotic yeast strains isolated from fermented food need to meet safe and beneficial conditions for the host's health. The Pichia kudriavzevii YGM091 strain isolated from fermented goat milk has outstanding probiotic characteristics, including: the high survival percentage in digestive system conditions (reaching up 247.13 ± 0.12 and 145.03 ± 0.06% at pH 3.0 and bile salt 0.5%, respectively); good tolerance to temperature, salt, phenol, ethanol; good surface properties such as high hydrophobicity percentage (> 60%), the high auto-aggregation percentage rate (66.56 ± 1.45% after 45 min of incubation) and the high co-aggregation percentage rate with pathogenic bacteria in a short time (> 40% after 2 h of incubation); biofilm forming after 24 h of incubation on abiotic surfaces; antioxidant activity reached excellent level after only 24 h of incubation (The percentage free radical scavenging and the Trolox equivalent reaching up 79.86 ± 0.70% and 92.09 ± 0.75 µg/mL after 72 h of incubation); extracellular enzymes production protease and cellulase with high activity, amylase and pectinase with moderate activity and non-lipase activity. Simultaneously, the YGM091 strain is the in vitro safety yeast: insensitive to antibiotics and fluconazole, negative for gelatinase, phospholipase, coagulase, and non-hemolysis activities. Furthermore, this strain is in vivo safety yeast with the dosages below 10 CFU/larva in the Galleria mellonella model with over 90% survival larvae and the yeast density reduced to just 10-10 CFU/larva after 72 h post-injection. Research results have demonstrated that the Pichia kudriavzevii YGM091 strain is a safe potential probiotic yeast and could become a candidate probiotic food to be used in the future.
PubMed: 37368223
DOI: 10.1007/s12602-023-10114-1 -
Scientific Reports Nov 2023Second-generation bioethanol production using lignocellulosic biomass as feedstock requires a highly efficient multistress-tolerant yeast. This study aimed to develop a...
Adaptive laboratory evolution under acetic acid stress enhances the multistress tolerance and ethanol production efficiency of Pichia kudriavzevii from lignocellulosic biomass.
Second-generation bioethanol production using lignocellulosic biomass as feedstock requires a highly efficient multistress-tolerant yeast. This study aimed to develop a robust yeast strain of P. kudriavzevii via the adaptive laboratory evolution (ALE) technique. The parental strain of P. kudriavzevii was subjected to repetitive long-term cultivation in medium supplemented with a gradually increasing concentration of acetic acid, the major weak acid liberated during the lignocellulosic pretreatment process. Three evolved P. kudriavzevii strains, namely, PkAC-7, PkAC-8, and PkAC-9, obtained in this study exhibited significantly higher resistance toward multiple stressors, including heat, ethanol, osmotic stress, acetic acid, formic acid, furfural, 5-(hydroxymethyl) furfural (5-HMF), and vanillin. The fermentation efficiency of the evolved strains was also improved, yielding a higher ethanol concentration, productivity, and yield than the parental strain, using undetoxified sugarcane bagasse hydrolysate as feedstock. These findings provide evidence that ALE is a practical approach for increasing the multistress tolerance of P. kudriavzevii for stable and efficient second-generation bioethanol production from lignocellulosic biomass.
Topics: Acetic Acid; Cellulose; Ethanol; Saccharomyces cerevisiae; Biomass; Furaldehyde; Saccharum; Fermentation
PubMed: 38017261
DOI: 10.1038/s41598-023-48408-7 -
Biotechnology For Biofuels 2020Although bioethanol production has been gaining worldwide attention as an alternative to fossil fuel, ethanol productivities and yields are still limited due to the...
BACKGROUND
Although bioethanol production has been gaining worldwide attention as an alternative to fossil fuel, ethanol productivities and yields are still limited due to the susceptibility of fermentation microorganisms to various stress and inhibitory substances. There is therefore an unmet need to search for multi-stress-tolerant organisms to improve ethanol productivity and reduce production cost, particularly when lignocellulosic hydrolysates are used as the feedstock.
RESULTS
Here, we have characterized a previously isolated LC375240 strain which is thermotolerant to high temperatures of 37 °C and 42 °C. More excitingly, growth and ethanol productivity of this strain exhibit strong tolerance to multiple stresses such as acetic acid, furfural, formic acid, HO and high concentration of ethanol at 42 °C. In addition, simple immobilization of LC375240 on corncobs resulted to a more stable and higher efficient ethanol production for successive four cycles of repeated batch fermentation at 42 °C.
CONCLUSION
The feature of being thermotolerant and multi-stress-tolerant is unique to LC375240 and makes it a good candidate for second-generation bioethanol fermentation as well as for investigating the molecular basis underlying the robust stress tolerance. Immobilization of LC375240 on corncobs is another option for cheap and high ethanol productivity.
PubMed: 32477425
DOI: 10.1186/s13068-020-01729-5 -
Mycotoxin Research Nov 2017The aim of this study was to evaluate the efficacy of autochthonous Pichia kudriavzevii as a novel bioadsorbent for aflatoxin B (AFB). The selection of this yeast was...
The aim of this study was to evaluate the efficacy of autochthonous Pichia kudriavzevii as a novel bioadsorbent for aflatoxin B (AFB). The selection of this yeast was based on the AFB adsorption capacity previously demonstrated in vitro (Magnoli et al. 2016). One-day-old Cobb broilers (n = 160) were randomly assigned to four dietary treatments (T1: basal diet (B); T2: B + 0.1% yeast; T3: B + AFB, 100 μg/kg; T4: B + 0.1% yeast + AFB, 100 μg/kg). Performance parameters (average daily weight gain body, average daily consumption, feed conversion ratio, carcass weight, and dead weight), biochemical parameters (albumin, globulin, and albumin/globulin), liver pathological changes, and AFB residual levels in the liver and excreta were evaluated. Significant differences (P < 0.05) in performance parameters were observed among treatments and controls: T3 group showed the lowest average daily body weight gain value while in T4 group, the value of this parameter increased significantly (P < 0.05). T3 and T4 groups showed the lowest and highest values for average daily feed consumption, respectively. The feed conversion ratio (FC) showed no significant differences among treatments. T3 group showed the lowest dead weight and carcass weight compared with T1 group. The biochemical parameters showed no significant differences among treatments. T3 group showed macroscopic and microscopic liver changes compared to the control. Aflatoxin B levels (μg/g) were detected in broiler livers and showed significant differences among treatments (P < 0.05). In conclusion, native P. kudriavzevii incorporation (0.1%) in broiler diets containing AFB was shown to be effective in ameliorating the adverse effects of AFB on production.
Topics: Aflatoxin B1; Animal Feed; Animals; Chickens; Diet; Dietary Supplements; Liver; Male; Pichia; Poultry Diseases; Random Allocation
PubMed: 28687999
DOI: 10.1007/s12550-017-0285-y -
Journal of Applied Microbiology May 2015Mineral deficiencies cause several health problems in the world, especially for populations consuming cereal-based diets rich in the anti-nutrient phytate. Our aim was...
AIMS
Mineral deficiencies cause several health problems in the world, especially for populations consuming cereal-based diets rich in the anti-nutrient phytate. Our aim was to characterize the phytate-degrading capacity of the yeast Pichia kudriavzevii TY13 and its secretion of phytase.
METHODS AND RESULTS
The phytase activity in cell-free supernatants from cultures with 100% intact cells was 35-190 mU ml(-1) depending on the media. The Km was 0.28 mmol l(-1) and the specific phytase activity 0.32 U mg(-1) total protein. The phytase activity and secretion of extracellular non-cell-bound phytase was affected by the medium phosphate concentrations. Further, addition of yeast extract had a clearly inducing effect, resulting in over 60% of the cultures total phytase activity as non-cell-bound.
CONCLUSIONS
Our study reveals that it is possible to achieve high extracellular phytase activity from the yeast P. kudriavzevii TY13 by proper composition of the growth medium.
SIGNIFICANCE AND IMPACT OF THE STUDY
TY13 could be a promising future starter culture for fermented foods with improved mineral bioavailability. Using strains that secrete phytase to the food matrix may significantly improve the phytate degradation by facilitating the enzyme-to-substrate interaction. The secreted non-cell-bound phytase activities by TY13 could further be advantageous in industrial production of phytase.
Topics: 6-Phytase; Extracellular Space; Fungal Proteins; Kinetics; Phytic Acid; Pichia
PubMed: 25630750
DOI: 10.1111/jam.12767 -
Applied Biochemistry and Biotechnology Aug 2023High-temperature ethanol fermentation (> 40 °C) can be applied as effective bioprocess technology to increase ethanol production. Thermotolerant yeast Pichia...
High-temperature ethanol fermentation (> 40 °C) can be applied as effective bioprocess technology to increase ethanol production. Thermotolerant yeast Pichia kudriavzevii 1P4 showed the ability to produce ethanol at optimum 37 °C. Thus, this study evaluated the ethanol productivity of isolate 1P4 at high-temperature ethanol fermentation (42 and 45 °C) and the identification of metabolite biomarkers using untargeted metabolomics with liquid chromatography-tandem mass spectrometry (LC-MS/MS). 1P4 showed tolerance to temperature stress up to 45 °C and thus relevant for high-temperature fermentation. As measured by gas chromatography (GC), bioethanol production of 1P4 at 30, 37, 42, and 45 °C was 5.8 g/l, 7.1 g/l, 5.1 g/l, and 2.8 g/l, respectively. The classification of biomarker compounds was based on orthogonal projection analysis to latent structure discriminant analysis (OPLS-DA), resulting in L-proline being a suspected biomarker compound for isolate 1P4 tolerance against high-temperature stress. Indeed, supplementation of L-proline on fermentation medium supported the growth of 1P4 at high temperatures (> 40 °C) than without L-proline. The bioethanol production with the addition of the L-proline resulted in the highest ethanol concentration (7.15 g/l) at 42 °C. Supplementation of L-proline as a stress-protective compound increased ethanol productivity at high-temperature fermentation of 42 and 45 °C by 36.35% and 83.33%, respectively, compared without the addition of L-proline. Preliminary interpretation of these results indicates that bioprocess engineering through supplementation of stress-protective compounds L-proline increases the fermentation efficiency of isolate 1P4 at higher temperatures (42 °C and 45 °C).
Topics: Fermentation; Temperature; Chromatography, Liquid; Tandem Mass Spectrometry; Gas Chromatography-Mass Spectrometry; Pichia; Yeasts; Ethanol
PubMed: 37103737
DOI: 10.1007/s12010-023-04554-2 -
International Journal of Molecular... Sep 2023Studying the production of Iron (Fe) nanoparticles using natural substances is an intriguing area of research in nanotechnology, as these nanoparticles possess...
Studying the production of Iron (Fe) nanoparticles using natural substances is an intriguing area of research in nanotechnology, as these nanoparticles possess biocompatibility and natural stability, which make them useful for a variety of industrial applications. The study utilized Fe nanoparticles that were synthesized using a bioflocculant and applied to eliminate different kinds of pollutants and dyes found in wastewater and solutions. The study involved the generation of Fe nanoparticles through a bioflocculant obtained from , which were evaluated for their flocculation and antimicrobial capabilities. The impact of the Fe nanoparticles on human embryonic kidney (HEK 293) cell lines was studied to assess their potential cytotoxicity effects. An array of spectroscopic and microscopic methods was employed to characterize the biosynthesized Fe nanoparticles, including SEM-EDX, FT-IR, TEM, XRD, UV-vis, and TGA. A highly efficient flocculating activity of 85% was achieved with 0.6 mg/mL dosage of Fe nanoparticles. The biosynthesized Fe nanoparticles demonstrated a noteworthy concentration-dependent cytotoxicity effect on HEK 293 cell lines with the highest concentration used resulting in 34% cell survival. The Fe nanoparticles exhibited strong antimicrobial properties against a variety of evaluated Gram-positive and Gram-negative microorganisms. The efficiency of removing dyes by the nanoparticles was found to be higher than 65% for the tested dyes, with the highest being 93% for safranine. The Fe nanoparticles demonstrated remarkable efficiency in removing various pollutants from wastewater. In comparison to traditional flocculants and the bioflocculant, biosynthesized Fe nanoparticles possess significant potential for eliminating both biological oxygen demand (BOD) and chemical oxygen demand (COD) from wastewater samples treated. Hence, the Fe nanoparticles synthesized in this way have the potential to substitute chemical flocculants in the treatment of wastewater.
Topics: Humans; Wastewater; Saccharomyces cerevisiae; Kombucha Tea; Iron; Spectroscopy, Fourier Transform Infrared; HEK293 Cells; Nanoparticles; Flocculation; Anti-Infective Agents; Environmental Pollutants; Coloring Agents; Hydrogen-Ion Concentration
PubMed: 37834177
DOI: 10.3390/ijms241914731 -
Foods (Basel, Switzerland) Feb 2023This study explored the effect of the combination of yeast, non- yeast (), and during cider fermentation on physicochemical properties, antioxidant activities, flavor...
This study explored the effect of the combination of yeast, non- yeast (), and during cider fermentation on physicochemical properties, antioxidant activities, flavor and aroma compounds, as well as sensory qualities. Ciders fermented with the triple mixed-cultures of these three species showed lower acid and alcohol content than those fermented with the single-culture of . The antioxidant activities were enhanced by the triple mixed-culture fermentation, giving a higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging rate and total antioxidant capacity; specifically, the SPL5 cider showed the highest DPPH radical scavenging rate (77.28%), while the SPL2 gave the highest total antioxidant capacity (39.57 mmol/L). Additionally, the triple mixed-culture fermentation resulted in improved flavor and aroma with a lower acidity (L-malic acid) and higher aroma compounds (Esters), when compared with the single-culture fermented ciders (); more specifically, the SPL4 cider resulted in the highest total flavor and aroma compounds. In addition, sensory evaluation demonstrated that ciders produced using the triple mixed-cultures gained higher scores than those fermented using the single-culture of , giving better floral aroma, fruity flavor, and overall acceptability. Therefore, our results indicated that the triple mixed-cultures (, , and ) were found to make up some enological shortages of the single fermented cider. This study is believed to provide a potential strategy to enhance cider quality and further give a reference for new industrial development protocols for cider fermentation that have better sensory qualities with higher antioxidant properties.
PubMed: 36766182
DOI: 10.3390/foods12030655 -
Eukaryotic Cell Oct 2012A draft genome sequence of Pichia kudriavzevii M12 is presented here. The genome reveals the presence of genes encoding enzymes involved in xylose utilization and the...
A draft genome sequence of Pichia kudriavzevii M12 is presented here. The genome reveals the presence of genes encoding enzymes involved in xylose utilization and the pentose phosphate pathway for bioethanol production. Strain M12 is also a potential producer of phytases, enzymes useful in food processing and agriculture.
Topics: 6-Phytase; Base Sequence; Ethanol; Genome, Fungal; Molecular Sequence Data; Pichia; Sequence Analysis, DNA
PubMed: 23027839
DOI: 10.1128/EC.00229-12