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Journal, Genetic Engineering &... Sep 2022Fermented foods are the results of metabolic activities of various microorganisms. People have traditionally known how to culture desirable microorganisms, primarily...
BACKGROUND
Fermented foods are the results of metabolic activities of various microorganisms. People have traditionally known how to culture desirable microorganisms, primarily lactic acid bacteria, yeasts, and filamentous molds, for the manufacture of edible foods. Yeast isolated from home-made mango pickle from Hamirpur, Himachal Pradesh, was assessed for probiotic properties and their enzymatic profiling.
RESULTS
Four yeast isolates were isolated out of which P. kudriavzevii Y33 was selected on the basis of high acid tolerance as well as broadest antimicrobial activity. The selected isolate was observed to have high acid tolerance at pH 2 and show strong antimicrobial activity against all the pathogens examined. P. kudriavzevii Y33 can also withstand high bile concentration and showed high viability index, i.e., 95% at concentration of 2% of bile. The isolate was able to demonstrate high cholesterol assimilation in medium containing ox bile and taurocholate, at 88.58 and 86.83%, respectively. The autoaggregation ability of isolate increases with increasing the time of incubation and showed 87% of autoaggregation after 24 h of incubation. P. kudriavzevii Y33 exhibited resistance towards different antibiotics, found to be positive for exopolysaccharide production and showed no hemolytic activity. The isolate was observed to produce several enzymes such as β-galactosidase, protease, amylase, phytase, and lipase.
CONCLUSIONS
The results of the current study revealed that P. kudriavzevii Y33 has various beneficial qualities that suggest it could be used as probiotics. Enzymes produced by yeast isolate help in improving flavor and mineral availability in the fermented products.
PubMed: 36083419
DOI: 10.1186/s43141-022-00416-2 -
Food Chemistry: X Jun 2023Mixed fermentation using saccharomyces cerevisiae and non- has become one of the main research strategies to improve wine aroma. Hence, this study applied the mixed...
Mixed fermentation using saccharomyces cerevisiae and non- has become one of the main research strategies to improve wine aroma. Hence, this study applied the mixed fermentation technique using and to brew Cabernet Sauvignon wine and to investigate the effects of inoculation timing and inoculation ratio on the polyphenolics, antioxidant activity and aroma of the resulting wine. The results showed that mixed fermentation significantly improved the amounts of flavan-3-ols. In particular, S1:5 had the highest amounts of (-)-catechin and procyanidin B1 (73.23 mg/L and 46.59 mg/L), while S1:10 had the highest (-)-epicatechin content (57.95 mg/L). Meanwhile, S1:10 showed the strongest FRAP, CUPRAC and ABTS + activities (31.46 %, 25.38 % and 13.87 % higher than that of CK, respectively). In addition, mixed fermentation also increased the amounts of phenylethanol, isoamyl alcohol and ethyl esters, which enhanced the rose-like and fruity flavor of wine. This work used a friendly non- alongside appropriate inoculation strategies to provide an alternative approach for improved wine aroma and phenolic profile.
PubMed: 37131849
DOI: 10.1016/j.fochx.2023.100685 -
Food Chemistry Feb 2021The use of starters during fermentation has been gaining momentum as it can warrant high-quality chocolate. The objective of this study was to investigate the influence...
The use of starters during fermentation has been gaining momentum as it can warrant high-quality chocolate. The objective of this study was to investigate the influence of Saccharomyces cerevisiae (Sc) and Pichia kudriavzevii (Pk) during on-farm fermentation on physico-chemical and microbiological characteristics and levels of methylxanthines and bioactive amines of cocoa. Four treatments were used: ScPk (1:1), only Sc, only Pk, and no starter (control). The starters lead to changes throughout fermentation, but provided fermented cocoa with similar pH, titratable acidity, reducing sugars and phenolic compounds. ScPk shortened fermentation time by 24 h. The ScPk fermented and dried cocoa had higher levels of monomeric phenols, methylxanthines, phenylethylamine and lower levels of the putrefactive amines - putrescine and cadaverine (p < 0.05). The results were confirmed by multivariate analysis. Based on these results, the mixture of both yeasts species is a promising starter for cocoa fermentation decreasing duration time and modulating high-quality components.
Topics: Amines; Cacao; Caffeine; Catechin; Chocolate; Farms; Fermentation; Food Microbiology; Hydrogen-Ion Concentration; Phenols; Pichia; Saccharomyces cerevisiae; Temperature; Theobromine; Theophylline; Time Factors
PubMed: 32810810
DOI: 10.1016/j.foodchem.2020.127834 -
Current Microbiology Oct 2023The isolation of endogenous yeast strains from traditionally fermented food products to use as functional starter cultures has become more popular for improved food...
The isolation of endogenous yeast strains from traditionally fermented food products to use as functional starter cultures has become more popular for improved food safety, quality, and beneficial health effects. In this study, 107 Pichia kudriavzevii strains were isolated from sourdough, shalgam, tarhana, artisanal Tulum cheese, and yogurt. The strains were identified by DNA fingerprinting using iPBS-PCR method before technological and probiotic characterization. The multivariate statistical approach revealed that five strains were most promising in terms of technological characterization, including different harsh growth conditions. These strains were also examined in terms of probiotic properties with a commercial S. cerevisiae var boulardii MYA-796 strain. The multivariate statistical analyses indicated that P. kudriavzevii 5S5 were most promising in in vitro probiotic properties such as surviving in human GI conditions, adhering to intestinal cell lines, and exhibiting high hydrophobicity. Therefore, it seems to be a great starter candidate for the production of functional fermented food products.
Topics: Humans; Saccharomyces cerevisiae; Fermentation; Probiotics; Fermented Foods
PubMed: 37861932
DOI: 10.1007/s00284-023-03505-8 -
Bioresource Technology Apr 2013D-xylonic acid is one of the top 30 most desirable chemicals to be derived from biomass sugars identified by the US Department of Energy, being applicable as a non-food...
D-xylonic acid is one of the top 30 most desirable chemicals to be derived from biomass sugars identified by the US Department of Energy, being applicable as a non-food substitute for D-gluconic acid and as a platform chemical. We engineered the non-conventional yeast Pichia kudriavzevii VTT C-79090T to express a D-xylose dehydrogenase coding gene from Caulobacter crescentus. With this single modification the recombinant P. kudriavzevii strain produced up to 171 g L(-1) of D-xylonate from 171 g L(-1) D-xylose at a rate of 1.4 g L(-1) h(-1) and yield of 1.0 g [g substrate consumed](-1), which was comparable with D-xylonate production by Gluconobacter oxydans or Pseudomonas sp. The productivity of the strain was also remarkable at low pH, producing 146 g L(-1) D-xylonate at 1.2 g L(-1) h(-1) at pH 3.0. This is the best low pH production reported for D-xylonate. These results encourage further development towards industrial scale production.
Topics: Biomass; Glucose; Hydrogen-Ion Concentration; Intracellular Space; Kluyveromyces; Pichia; Saccharomyces cerevisiae; Sugar Acids; Xylitol; Xylose
PubMed: 23455228
DOI: 10.1016/j.biortech.2013.01.157 -
Journal of Fungi (Basel, Switzerland) Dec 2021Improving the comprehensive utilization of sugars in lignocellulosic biomass is a major challenge for enhancing the economic viability of lignocellulose biorefinement. A...
Improving the comprehensive utilization of sugars in lignocellulosic biomass is a major challenge for enhancing the economic viability of lignocellulose biorefinement. A robust yeast N-X showed excellent performance in ethanol production under high temperature and low pH conditions and was engineered for ᴅ-xylonate production without xylitol generation. The recombinant strain N-X/S1 was employed for sequential production of ᴅ-xylonate and ethanol from ᴅ-xylose, feeding on ᴅ-glucose without pH control in a two-stage strategy of aerobic and shifting micro-aerobic fermentation. Acid-pretreated corncob without detoxification and filtration was used for ᴅ-xylonate production, then simultaneous saccharification and ethanol fermentation was performed with cellulase added at pH 4.0 and at 40 °C. By this strategy, 33.5 g/L ᴅ-xylonate and 20.8 g/L ethanol were produced at yields of 1.10 g/g ᴅ-xylose and 84.3% of theoretical value, respectively. We propose a promising approach for the sequential production of ᴅ-xylonate and ethanol from non-detoxified corncob using a single microorganism.
PubMed: 34947020
DOI: 10.3390/jof7121038 -
AMB Express May 2018The yeast strain SJP-SNU was investigated as a probiotic and was characterized with respect to growth temperature, bile salt resistance, hydrogen sulfide reducing...
The yeast strain SJP-SNU was investigated as a probiotic and was characterized with respect to growth temperature, bile salt resistance, hydrogen sulfide reducing activity, intestinal survival ability and chicken embryo pathogenicity. In addition, we determined the complete genomic and mitochondrial sequences of SJP-SNU and conducted comparative genomics analyses. SJP-SNU grew rapidly at 37 °C and formed colonies on MacConkey agar containing bile salt. SJP-SNU reduced hydrogen sulfide produced by Salmonella serotype Enteritidis and, after being fed to 4-week-old chickens, could be isolated from cecal feces. SJP-SNU did not cause mortality in 10-day-old chicken embryos. From 13 initial contigs, 11 were finally assembled and represented 10 chromosomal sequences and 1 mitochondrial DNA sequence. Comparative genomic analyses revealed that SJP-SNU was a strain of Pichia kudriavzevii. Although SJP-SNU possesses pathogenicity-related genes, they showed very low amino acid sequence identities to those of Candida albicans. Furthermore, SJP-SNU possessed useful genes, such as phytases and cellulase. Thus, SJP-SNU is a useful yeast possessing the basic traits of a probiotic, and further studies to demonstrate its efficacy as a probiotic in the future may be warranted.
PubMed: 29774473
DOI: 10.1186/s13568-018-0609-0 -
Microbial Biotechnology May 2021The unconventional yeast Pichia kudriavzevii is renowned for its ability to survive at low pH and has been exploited for the industrial production of various organic...
The unconventional yeast Pichia kudriavzevii is renowned for its ability to survive at low pH and has been exploited for the industrial production of various organic acids, especially succinic acid (SA). However, P. kudriavzevii can also utilize the di- and tricarboxylate intermediates of the Krebs cycle as the sole carbon sources for cell growth, which may adversely affect the extracellular accumulation of SA. Because the carboxylic acid transport machinery of P. kudriavzevii remains poorly understood, here, we focused on studying its SA transportation process from the perspective of mining and characterization of dicarboxylate transporters in a newly isolated acid-tolerant P. kudriavzevii strain CY902. Through genome sequencing and transcriptome analysis, two JEN family carboxylate transporters (PkJEN2-1 and PkJEN2-2) were found to be involved in SA transport. Substrate specificity analysis revealed that both PkJEN proteins are active dicarboxylate transporters, that can effectively import succinate, fumarate and L-malate into the cell. In addition, PkJEN2-1 can transport α-ketoglutarate, while PkJEN2-2 cannot. Since PkJEN2-1 shows higher transcript abundance than PkJEN2-2, its role in dicarboxylate transport is more important than PkJEN2-2. In addition, PKJEN2-2 is also responsible for the uptake of citrate. To our best knowledge, this is the first study to show that a JEN2 subfamily transporter is involved in tricarboxylate transport in yeast. A combination of model-based structure analysis and rational mutagenesis further proved that amino acid residues 392-403 of the tenth transmembrane span (TMS-X) of PkJEN2-2 play an important role in determining the specificity of the tricarboxylate substrate. Moreover, these two PkJEN transporters only exhibited inward transport activity for SA, and simultaneous inactivation of both PkJEN transporters reduced the SA influx, resulting in enhanced extracellular accumulation of SA in the late stage of fermentation. This work provides useful information on the mechanism of di-/tricarboxylic acid utilization in P. kudriavzevii, which will help improve the organic acid production performance of this microbial chassis.
Topics: Membrane Transport Proteins; Pichia; Saccharomyces cerevisiae; Succinates; Succinic Acid
PubMed: 33629807
DOI: 10.1111/1751-7915.13781 -
Food Science and Biotechnology Apr 2021Co-fermentation using yeast ( and ) and the bacteria () as starters isolated from spontaneous sourdough was conducted for the brewing of glucuronic acid (GlcA)-enriched...
Co-fermentation using yeast ( and ) and the bacteria () as starters isolated from spontaneous sourdough was conducted for the brewing of glucuronic acid (GlcA)-enriched apple cider. The concentration of GlcA in the apple cider co-fermented for 14 d with commercial and was 37.7 ± 1.7 mg/mL while a concentration of 62.8 ± 3.1 mg/mL was recorded for fermentation with and , which was higher than the corresponding single yeast fermentation. The co-fermented apple cider revealed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of 171.67 ± 0.79 µg trolox equivalents (TE)/mL using and , compared to the control (143.89 ± 7.07 µg TE/mL) just using . Thus, the co-fermentation of and and and provided a new strategy for the development of GlcA-enriched apple cider with enhanced antioxidant capacity.
PubMed: 33936847
DOI: 10.1007/s10068-021-00883-2 -
Biotechnology For Biofuels Nov 2021High-temperature bioethanol production benefits from yeast thermotolerance. Salt stress could induce obvious cross-protection against heat stress of Pichia kudriavzevii,...
Salt stress improves thermotolerance and high-temperature bioethanol production of multi-stress-tolerant Pichia kudriavzevii by stimulating intracellular metabolism and inhibiting oxidative damage.
BACKGROUND
High-temperature bioethanol production benefits from yeast thermotolerance. Salt stress could induce obvious cross-protection against heat stress of Pichia kudriavzevii, contributing to the improvement of its thermotolerance and bioethanol fermentation. However, the underlying mechanisms of the cross-protection remain poorly understood.
RESULTS
Salt stress showed obvious cross-protection for thermotolerance and high-temperature ethanol production of P. kudriavzevii observed by biomass, cell morphology and bioethanol production capacity. The biomass and ethanol production of P. kudriavzevii at 45 °C were, respectively, improved by 2.6 and 3.9 times by 300 mmol/L NaCl. Metabolic network map showed that salt stress obviously improved the key enzymes and intermediates in carbohydrate metabolism, contributing to the synthesis of bioethanol, ATP, amino acids, nucleotides, and unsaturated fatty acids, as well as subsequent intracellular metabolisms. The increasing trehalose, glycerol, HSPs, and ergosterol helped maintain the normal function of cell components. Heat stress induced serious oxidative stress that the ROS-positive cell rate and dead cell rate, respectively, rose from 0.5% and 2.4% to 28.2% and 69.2%, with the incubation temperature increasing from 30 to 45 °C. The heat-induced ROS outburst, oxidative damage, and cell death were obviously inhibited by salt stress, especially the dead cell rate which fell to only 20.3% at 300 mmol/L NaCl. The inhibiting oxidative damage mainly resulted from the abundant synthesis of GSH and GST, which, respectively, increased by 4.8 and 76.1 times after addition of 300 mmol/L NaCl. The improved bioethanol production was not only due to the improved thermotolerance, but resulted from the up-regulated alcohol dehydrogenases and down-regulated aldehyde dehydrogenases by salt stress.
CONCLUSION
The results provide a first insight into the mechanisms of the improved thermotolerance and high-temperature bioethanol production of P. kudriavzevii by salt stress, and provide important information to construct genetic engineering yeasts for high-temperature bioethanol production.
PubMed: 34823567
DOI: 10.1186/s13068-021-02071-0