• Journal of Microbiology and Biotechnology
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• v.33 no.11
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• pp.1506-1512
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• 2023

Quantitative analysis of adenosine triphosphate (ATP) has been widely used as a diagnostic tool in the food and medical industries. Particularly, the pathogenesis of a few diseases including inflammatory bowel disease (IBD) is closely related to high ATP concentrations. A bioluminescent D-luciferin/luciferase system, which includes a luciferase (FLuc) from the firefly Photinus pyralis as a key component, is the most commonly used method for the detection and quantification of ATP. Here, instead of isolating FLuc produced in recombinant Escherichia coli, we aimed to develop a whole-cell biocatalyst system that does not require extraction and purification of FLuc. To this end, the gene coding for FLuc was introduced into the genome of probiotic Saccharomyces boulardii using the CRISPR/Cas9-based genome editing system. The linear relationship (r² = 0.9561) between ATP levels and bioluminescence generated from the engineered S. boulardii expressing FLuc was observed in vitro. To explore the feasibility of using the engineered S. boulardii expressing FLuc as a whole-cell biosensor to detect inflammation biomarker (i.e., ATP) in the gut, a colitis mouse model was established using dextran sodium sulfate as a colitogenic compound. Our findings demonstrated that the whole-cell biosensor can detect elevated ATP levels during gut inflammation in mice. Therefore, the simple and powerful method developed herein could be applied for non-invasive IBD diagnosis.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.744-750
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• 1999

Forty alcohol yeast strains were isolated from the main mashes (10 strains from each mash) for brewing of 4 different kinds of Korean traditional liquor (3 different types of Yakju and 1 Takju). Thirty-eight out of 40 strains were identified to be the same strain, Saccharomyces boulardii, by the Automated Bacteria, Yeast, and Fungi Identification System (Biolog Co., U.S.A.) based on the metabolic fingerprints. One strain that showed the highest ethanol production among the 38 strains in YPD medium, designated SHY 111, was selected and used for differentiating from other yeast type strains using the polymerase chain reaction (PCR). Amplified DNA, from transcribed internal spacers of SHY 111 chromosomal DNA, was found to be the same in both size and sequence as those of S. cerevisiae KCCM 11215 (formerly S. coreanus) and S. boulardii along with that of S. cerevisiae AB 972, which was used as a type strain for the yeast genome project. However, when PCR was carried out with the intron splice site primer, it resulted in the amplification of the SHY 111-specific DNA fragment which was about 200 bp in size. When PCR was carried out using the primer to test diversity of 40 isolated yeast strains, it was found that the PCR patterns were similar to each other except for the 200 bp bands derived from all the 10 strains from one Yakju, and 2 strains from another Yakju. These results suggest the strain identified as S. boulardii by the Automated Identification System to be a dominant strain for the fermentation of Korean traditional liquors.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.18 no.1
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• pp.17-22
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• 2015

Purpose: This study aims to investigate Saccharomyces boulardii CNCM I-745 during Helicobacter pylori eradication in children. Methods: One hundred ninety-four H. pylori positive children were randomized in two groups. Therapy (omeprazole+ clarithromycin+amoxicillin or omeprazole+clarithromycin+metronidazole in case of penicillin allergy) was given to both groups during two weeks. In the treatment group (n: 102) S. boulardii was added to the triple therapy, while the control group (n: 92) only received triple therapy. The incidence, onset, duration and severity of diarrhea and compliance to the eradication treatment were compared. A $^{13}C$ urea breath test was done 4 weeks after the end of eradication therapy in two groups of 21 patients aged 12 years and older to test the H. pylori eradication rate. Results: In the treatment group, diarrhea occurred in 12 cases (11.76%), starting after $6.25{\pm}1.24days$, lasting $3.17{\pm}1.08days$, and compliance to eradication treatment was 100%. In the control group, diarrhea occurred in 26 cases (28.26%), starting after $4.05{\pm}1.11days$, lasting $4.02{\pm}0.87days$, and in six cases eradication treatment was stopped prematurely (p<0.05). The $^{13}C$ urea breath test showed successful H. pylori eradication in 71.4% of the patients in the treatment and in 61.9 % in the control group (not significant). Conclusion: S. boulardii has a beneficial effect on the prevention and treatment of diarrhea during H. pylori eradication in children. Although S. boulardii did only slightly increase H. pylori eradication rate, compliance to eradication treatment was improved.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.54-61
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• 2020

Saccharomyces boulardii is the only probiotic yeast with US Food and Drug Administration approval. It is routinely used to prevent or treat acute diarrhea and other gastrointestinal disorders, including the antibiotic-associated diarrhea caused by Clostridium difficile infections. The formation of reactive oxygen species (ROS), specifically H₂O₂ during normal aerobic metabolism, contributes to programmed cell death and represents a risk to the viability of the probiotic microbe. Moreover, a loss of viability reduces the efficacy of the probiotic treatment. Therefore, inhibiting the accumulation of ROS in the oxidant environment could improve the viability of the probiotic yeast and lead to more efficacious treatment. Here, we provide evidence that supplementation with a non-reducing disaccharide, namely trehalose, enhanced the viability of S. boulardii exposed to an oxidative environment by preventing metacaspase YCA1-mediated programmed cell death through inhibition of intracellular ROS production. Our results suggest that supplementation with S. boulardii together with trehalose could increase the viability of the organism, and thus improve its effectiveness as a probiotic and as a treatment for acute diarrhea and other gastrointestinal disorders.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.24 no.5
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• pp.423-431
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• 2021

Purpose: To assess the effect of combination probiotic Saccharomyces boulardii CNCM-I 3799 and Bacillus subtilis CU-1 in outpatient management of acute watery diarrhea in children. Methods: A randomized double-blind placebo-controlled study was conducted in 180 participants aged six months to five years with acute mild to moderate diarrhea. All were enrolled from six centers across India and centrally randomized to receive S. boulardii CNCM-I 3799 and B. subtilis CU-1 or a placebo along with oral rehydration salts and zinc supplementation. Each participant was followed up for three months to assess recurrence of diarrhea. Results: The mean duration of diarrhea in the probiotic and placebo groups were 54.16 hours and 59.48 hours, respectively. The difference in the duration of diarrhea in those administered with probiotic or placebo within 24 hours of diarrhea onset was 25.21 hours. Furthermore, the difference in duration of diarrhea was 13.84 hours (p<0.05) for participants who were administered with probiotics within 48 hours. There were no significant differences in the stool frequencies between the two arms. After three months, 15% in the probiotic group and 18.5% in the placebo group reported episodes of diarrhea. The mean duration of diarrhea was considerably lower in the probiotic group, 31.02 hours versus 48 hours in placebo (p=0.017). Conclusion: S. boulardii CNCM-I 3799 and B. subtilis CU-1 combination was effective in reducing the duration of diarrhea when administered within 48 hours of diarrhea onset. Similarly, it reduced recurrence of diarrhea and its intensity in the subsequent three months.

• Journal of Microbiology and Biotechnology
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• v.33 no.10
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• pp.1329-1336
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• 2023

Microbial fermentation is often used to improve the functionality of plant-based food materials. Herein, we investigated changes in the physicochemical and functional properties of cabbage during yeast fermentation to develop new products using fermented cabbage. Among the 8 types of food-grade yeast, both Saccharomyces cerevisiae and Saccharomyces boulardii fermented 10% cabbage powder solution (w/w) the most effectively, leaving no soluble sugars after 12 h of fermentation. In addition, the yeast fermentation of cabbage resulted in functionally positive outcomes in terms of sulforaphane content, antioxidant properties, and anti-inflammatory activity. Specifically, the yeast-fermented cabbages contained about 500% more sulforaphane. The soluble fraction (5 ㎍/ml) of yeast-fermented cabbage had no cytotoxicity in murine RAW 264.7 cells, and the radical-scavenging capacity was equivalent to 1 ㎍/ml of ascorbic acid. Moreover, cabbage fermented with S. boulardii significantly suppressed both lipopolysaccharides (LPS)-induced nitric oxide production and LPS-induced reactive oxygen species production in RAW 264.7 cells, suggesting a potential anti-inflammatory effect. These results support the idea that yeast fermentation is promising for developing functionally improved cabbage products.

• Journal of Microbiology and Biotechnology
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• v.32 no.7
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• pp.877-884
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• 2022

Probiotics are microorganisms that can benefit host health when ingested in a live state, and lactic acid bacteria are the most common type. Among fungi, Saccharomyces boulardii (SB) is the only strain known to have a probiotic function with beneficial effects on colitis; however, information on other probiotic yeast strains is limited. Therefore, this study aimed to discover yeast strains expressing intestinal anti-inflammatory activities by exhibiting probiotic properties in dextran sodium sulfate (DSS)-induced colitis mice model. Nuruk (Korean traditional fermentation starter) containing various microbial strains was used as a source for yeast strains, and S. cerevisiae 28-7 (SC28-7) strain was selected with in vitro and in vivo characteristics to enable survival in the intestines. After 14 days of pretreatment with the yeast strains, DSS was co-administered for six days to induce colitis in mice. The results revealed that the disease activity index score was lowered by SC28-7 treatment compared to the DSS group, and the colon length and weight/length ratio were recovered in a pattern similar to that of the normal group. SC28-7 administration significantly reduced the secretion of pro-inflammatory cytokines in the serum and modified the mRNA expression of inflammatory cytokines (interleukin-1β, transforming growth factor-β, and interferon-γ) and proteins involved in gut barrier functions (mucin 2, mucin 3, zonula occludens-1, and occludin) in colon tissues. These results indicate that SC28-7 attenuates DSS-induced colon damage and inflammation, supporting its future use as a probiotic yeast for treating and preventing intestinal inflammatory diseases such as inflammatory bowel disease.

• Restorative Dentistry and Endodontics
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• v.47 no.3
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• pp.31.1-31.9
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• 2022

Objectives: This study aimed to evaluate in vitro the effects of the self-adhesive resin cements RelyX U200 (3M ESPE) and seT PP (SDI Limited) on murine macrophages and the interference of the photoactivation. Materials and Methods: Cell viability assays, cell adherence, yeast phagocytosis of Saccharomyces boulardii and production of reactive oxygen species (ROS) were performed in the presence of capillaries containing the respective self-adhesive cement when photoactivated or not. Results: After long periods of contact, both types of cements, when not photoactivated, are more cytotoxic for macrophages. The seT PP cement when only chemically activated seems to interfere more negatively in the process of phagocytosis of yeasts S. boulardii. Both types of cements interfere in the cell adhesion process, independent of photoactivation. None of the types of cements tested was able to induce the production of ROS. Conclusions: Our results highlight the great importance of the photoactivation of self-adhesive resin cements in the dental clinic, since RelyX U200, when photoactivated, presented the best results within the evaluated parameters.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.20 no.3
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• pp.139-146
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• 2017

The number of papers discussing probiotics increases tremendously that limits the possibility for primary care physicians and clinicians to stay updated. Therefore, the aim of this paper will be to summarize available evidence of probiotic use in well-defined clinical indications of importance for pediatricians. Based on currently available evidence certain probiotic strains (Lactobacillus rhamnosus GG [LGG] and Saccharomyces boulardii) have proven effect in the treatment of acute gastroenteritis and prevention of antibiotic associated diarrhea. Furthermore, LGG was proven to be effective in prevention of nosocomial diarrhea and respiratory tract infection in day care centers. In conclusion, not all probiotic strains have same efficacy for all clinical indications, therefore, only strains with proven efficacy and safety should be recommended.

• Microbiology and Biotechnology Letters
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• v.50 no.3
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• pp.361-374
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• 2022

For this study, we designed and produced mulberry low-fat ice cream supplemented with synbiotics (MLF-ISS). The sensory characteristics and physical, chemical, and microbiological qualities of MLF-ISS were then determined. Mulberry juice inoculated with or without probiotic (Lactobacillus plantarum TISTR 926 and Saccharomyces boulardii CNCM I-745) was also tested at 37℃ for 24 h to determine probiotic growth rate, pH, total anthocyanin content (TAC), total phenolic content (TPC), and antioxidant activity (AA). Only the TAC of mulberry juice inoculated with S. boulardii CNCM I-745 increased considerably (p < 0.05) among these parameters. MLF-ISS was produced with varied mulberry fruit concentrations (0, 10, 20, 30, or 40%) (w/w). The MLF-ISS prepared with 30% mulberry fruit (w/w) (30-MLF-ISS) had a higher score in appearance, color, and sweetness (p < 0.05) when sensory qualities were measured using the 9-point hedonic scale method. In the CIE lab system (L^*, a^*, b^*), the color values of 30-MLF-ISS were 27.80 ± 0.26, 12.99 ± 0.59, and 1.43 ± 0.05, respectively. The 30-MLF-ISS was also subjected to a proximate analysis. The melting rate of 30-MLF-ISS was 0.29 ± 0.03 g/min and the time it took for the first drop to fall was 37.00 ± 7.00 min. TAC, TPC, and AA of 30-MLF-ISS were observed to alter significantly (p < 0.05) during varied intervals of storage at - 18℃ (0, 30, and 60 days). The viability of probiotics in 30-MLF-ISS slightly decreased after storage at -18℃ for 8 weeks, but remained about 6 log CFU/g. During storage at -18℃ for 0 and 120 days, no pathogenic bacteria were detected in 30-MLF-ISS. These findings show that 30-MLF-ISS has nutritional and functional value, is free of foodborne pathogenic bacteria, is safe for consumers' health, and is suitable for application in the ice cream and related food industries.