• Title/Summary/Keyword: yeast strains

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Antimicrobial Activity of Kefir against Various Food Pathogens and Spoilage Bacteria

  • Kim, Dong-Hyeon;Jeong, Dana;Kim, Hyunsook;Kang, Il-Byeong;Chon, Jung-Whan;Song, Kwang-Young;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.36 no.6
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    • pp.787-790
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    • 2016
  • Kefir is a unique fermented dairy product produced by a mixture of lactic acid bacteria, acetic acid bacteria, and yeast. Here, we compared the antimicrobial spectra of four types of kefirs (A, L, M, and S) fermented for 24, 36, 48, or 72 h against eight food-borne pathogens. Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Enterococcus faecalis, Escherichia coli, Salmonella Enteritidis, Pseudomonas aeruginosa, and Cronobacter sakazakii were used as test strains, and antibacterial activity was investigated by the spot on lawn method. The spectra, potencies, and onsets of activity varied according to the type of kefir and the fermentation time. The broadest and strongest antimicrobial spectrum was obtained after at least 36-48 h of fermentation for all kefirs, although the traditional fermentation method of kefir is for 18-24 h at $25^{\circ}C$. For kefir A, B. cereus, E. coli, S. Enteritidis, P. aeruginosa, and C. sakazakii were inhibited, while B. cereus, S. aureus, E. coli, S. Enteritidis, P. aeruginosa, and C. sakazakii were inhibited to different extents by kefirs L, M, and S. Remarkably, S. aureus, S. Enteritidis, and C. sakazakii were only inhibited by kefirs L, M, and S, and L. monocytogenes by kefir M after fermentation for specific times, suggesting that the antimicrobial activity is attributable not only to a low pH but also to antimicrobial substances secreted during the fermentation.

Isolation of Fungal Deteriogens Inducing Aesthetical Problems and Antifungal Calcite Forming Bacteria from the Tunnel and Their Characteristics (터널에서 미학적 문제를 야기하는 진균 및 항진균 활성을 가진 탄산칼슘 형성세균의 분리와 특성)

  • Park, Jong-Myong;Park, Sung-Jin;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.287-293
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    • 2011
  • The purpose of this study was to isolate and characterize fungal deteriogens, which induce discoloration of the cement tunnel, and calcite forming bacteria (CFBs), which have antifungal activity against fungal deteriogens. Isolation of mold, bacteria and yeast was performed using several solid media and partially identified using internal transcribed spacer (ITS); 5.8S rRNA gene sequencing and 16s rDNA sequencing. A total of 19 microbial strains were identified with the most widely distributed fungal strain being Cladospirum sphaerospermum. In addition, five bacteria derived from the tunnel were identified as CFBs. Amongst the latter, Bacillus aryabhatti KNUC205 exhibited antifungal activity against Cladospirum sphaerospermum KNUC253 and Aspergillus niger KCTC6906 as concentrated filtered supernatants.

Fabrication of Biogenic Antimicrobial Silver Nanoparticles by Streptomyces aegyptia NEAE 102 as Eco-Friendly Nanofactory

  • El-Naggar, Noura El-Ahmady;Abdelwahed, Nayera A.M.;Darwesh, Osama M.M.
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.453-464
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    • 2014
  • The current research was focused on the extracellular biosynthesis of bactericidal silver nanoparticles (AgNPs) using cell-free supernatant of a local isolate previously identified as a novel Streptomyces aegyptia NEAE 102. The biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102 was quite fast and required far less time than previously published strains. The produced particles showed a single surface plasmon resonance peak at 400 nm by UV-Vis spectroscopy, which confirmed the presence of AgNPs. Response surface methodology was chosen to evaluate the effects of four process variables ($AgNO_3$ concentration, incubation period, pH levels, and inoculum size) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. Statistical analysis of the results showed that the linear and quadratic effects of incubation period, initial pH, and inoculum size had a significant effect (p < 0.05) on the biosynthesis of silver nanoparticles by Streptomyces aegyptia NEAE 102. The maximum silver nanoparticles biosynthesis (2.5 OD, at 400 nm ) was achieved in runs number 5 and 14 under the conditions of 1 mM $AgNO_3$ (1-1.5% (v/v)), incubation period (72-96 h), initial pH (9-10), and inoculum size (2-4% (v/v)). An overall 4-fold increase in AgNPs biosynthesis was obtained as compared with that of unoptimized conditions. The biosynthesized silver nanoparticles were characterized using UV-VIS spectrophotometer and Fourier transform infrared spectroscopy analysis, in addition to antimicrobial properties. The biosynthesized AgNPs significantly inhibited the growth of medically important pathogenic gram-positive (Staphylococcus aureus) and gram-negative bacteria (Pseudomonas aeruginosa) and yeast (Candida albicans).

A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

  • Leem, Sun-Hee
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.248-255
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    • 1999
  • We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

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The Inhibitory Effects of Chloroform Fraction Extracted from the Dandelion (Taraxacum platycarpum D.) against Lactic Acid Bacteria and Yeast Related to Kimchi Fermentation (포공영 클로로포름 분획의 김치 유산균 및 효모에 대한 저해 효과)

  • Kim, Jin-Hee;Kim, Mee-Ree
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.560-566
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    • 2001
  • This study was conducted to investigate the effects of solvent extracts from medicinal plants on the fermentation of kimchi. Five microorganisms related to kimchi fermentation were selected and the antimicrobial activities of solvent fractions from medicinal plants were investigated. The chloroform fraction from the methanol extract of dandelion (Taraxacum platycarpum D.) exhibited inhibitory activity against five strains such as Lactobacilli plantarum, Lactobacilli brevis, Leuconostoc mesenteroides, Enterococcus faecalis and Saccharomyces cerevisiae. The chloroform fraction from the methanol extract of Dandelion inhibited the growth of E. faecalis and Leu. mesenteroides at the concentration of 80 mg/mL. Scanning electron micrographs of Leu. mesenteroides and E. faecalis treated with chloroform fraction 80 mg/mL exhibited morphological changes, including irregularly contracted cell surface.

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Enhanced physiological activities of wine yeasts Pichia manshurica and Pichia terricola isolated from grapes pericarp (포도 과피에서 분리한 효모 Pichia manshurica와 Pichia terricola의 생리활성 증진효과)

  • Park, Sang-Kook;Kim, Dong-Min;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.235-241
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    • 2017
  • The purpose of this study was to investigate the enhanced physiological activities of two Pichia strains, yeasts isolated from grapes pericarp. Based on phylogenetic analysis using 18S rRNA sequencing, two isolates were identified as Pichia manshurica GU-3 and Pichia terricola GU-4, respectively. The scanning electron microscopic analysis showed that the two isolates grown on YPD medium were of typical elliptical shape with buds and bud scars on cell surface. Physiological activities of the single and mixed Pichia cultures were monitored and compared. In mixed cultures after 72 h of incubation, the maximum activities of tyrosinase inhibition, ACE inhibition, and antioxidant were 81.7%, 45.9%, and 42.7%, respectively. Superoxide dismutase-like activity was approximately 30% in the mixed cultures. These studies demonstrate that Pichia species cultured in the form of mixture can enhance the physiological activities and has potential for the development of new bioactive products.

Protection of Metal Stress in Saccharomyces cerevisiae: Cadmium Tolerance Requies the Presence if Two ATP-Binding Domains of Hsp 104 Protein

  • Lee, Gyeong Hui;Eom, Jeong Hun
    • Bulletin of the Korean Chemical Society
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    • v.22 no.5
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    • pp.514-518
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    • 2001
  • We have explored the importance of two ATP binding domains of Hsp104 protein in protection of yeast cells from cadmium exposure. In the previous study we have discovered that the presence of two ATP binding sites was essential in providing heat sh ock protection as well as rescuing cells from oxidative stress. In this paper we first report wild type cell with functional hsp104 gene is more resistant to cadmium stress than hsp104-deleted mutant cell, judging from decrease in survival rates as a result of cadmium exposure. In order to demonstrate functional role of two ATP binding sites in cadmium defense, we have transformed both wild type (SP1) and hyperactivated ras mutant (IR2.5) strains with several plasmids differing in the presence of ATP binding sites. When an extra copy of functional hsp104 gene with both ATP binding sites was overexpressed with GPD-promoter, cells showed increased survival rate against cadmium stress than mutants with ATP binding sites changed. The degree of protection in the presence of two ATP binding sites was similarly observed in ira2-deleted hyperactivated ras mutant, which was more sensitive to oxidative stress than wild type cell. We have concluded that the greater sensitivity to cadmium stress in the absence of two ATP binding sites is attributed to the higher concentration of reactive oxygen species (ROS) produced by cadmium exposure based on the fluorescence tests. These findings, taken all together, imply that the mechanism by which cadmium put forth toxic effects may be closely associated with the oxidative stress, which is regulated independently of the Ras-cAMP pathway. Our study provides a better understanding of cadmium defense itself and cross-talks between oxidative stress and metal stress, which can be applied to control human diseases due to similar toxic environments.

Heterologous Expression of Rhizopus Oryzae CYP509C12 Gene in Rhizopus Nigricans Enhances Reactive Oxygen Species Production and 11α-Hydroxylation Rate of 16α, 17-Epoxyprogesterone

  • Shen, Chaohui;Gao, Xiyang;Li, Tao;Zhang, Jun;Gao, Yuqian;Qiu, Liyou;Zhang, Guang
    • Mycobiology
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    • v.47 no.3
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    • pp.301-307
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    • 2019
  • The $11{\alpha}$-hydroxylation of $16{\alpha}$, 17-epoxyprogesterone (EP) catalyzed by Rhizopus nigricans is crucial for the steroid industry. However, lower conversion rate of the biohydroxylation restricts its potential industrial application. The $11{\alpha}$-steroid hydroxylase CYP509C12 from R. oryzae were reported to play a crucial role in the $11{\alpha}$-hydroxylation in recombinant fission yeast. In the present study, the CYP509C12 of R. oryzae (RoCYP) was introduced into R. nigricans using the liposome-mediated mycelial transformation. Heterologous expression of RoCYP resulted in increased fungal growth and improved intracellular reactive oxygen species content in R. nigricans. The $H_2O_2$ levels in RoCYP transformants were approximately 2-folder that of the R. nigricans wild type (RnWT) strain, with the superoxide dismutase activities increased approximately 45% and catalase activities decreased approximately 68%. Furthermore, the $11{\alpha}$-hydroxylation rates of EP in RoCYP transformants (C4, C6 and C9) were 39.7%, 38.3% and 38.7%, which were 12.1%, 8.2% and 9.4% higher than the rate of the RnWT strain, respectively. This paper investigated the effect of heterologous expression of RoCYP in R. nigricans, providing an effective genetic method to construct the engineered strains for steroid industry.

Methylglyoxal-Scavenging Enzyme Activities Trigger Erythroascorbate Peroxidase and Cytochrome c Peroxidase in Glutathione-Depleted Candida albicans

  • Kang, Sa-Ouk;Kwak, Min-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.79-91
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    • 2021
  • γ-Glutamylcysteine synthetase (Gcs1) and glutathione reductase (Glr1) activity maintains minimal levels of cellular methylglyoxal in Candida albicans. In glutathione-depleted Δgcs1, we previously saw that NAD(H)-linked methylglyoxal oxidoreductase (Mgd1) and alcohol dehydrogenase (Adh1) are the most active methylglyoxal scavengers. With methylglyoxal accumulation, disruptants lacking MGD1 or ADH1 exhibit a poor redox state. However, there is little convincing evidence for a reciprocal relationship between methylglyoxal scavenger genes-disrupted mutants and changes in glutathione-(in)dependent redox regulation. Herein, we attempt to demonstrate a functional role for methylglyoxal scavengers, modeled on a triple disruptant (Δmgd1/Δadh1/Δgcs1), to link between antioxidative enzyme activities and their metabolites in glutathione-depleted conditions. Despite seeing elevated methylglyoxal in all of the disruptants, the result saw a decrease in pyruvate content in Δmgd1/Δadh1/Δgcs1 which was not observed in double gene-disrupted strains such as Δmgd1/Δgcs1 and Δadh1/Δgcs1. Interestingly, Δmgd1/Δadh1/Δgcs1 exhibited a significantly decrease in H2O2 and superoxide which was also unobserved in Δmgd1/Δgcs1 and Δadh1/Δgcs1. The activities of the antioxidative enzymes erythroascorbate peroxidase and cytochrome c peroxidase were noticeably higher in Δmgd1/Δadh1/Δgcs1 than in the other disruptants. Meanwhile, Glr1 activity severely diminished in Δmgd1/Δadh1/Δgcs1. Monitoring complementary gene transcripts between double gene-disrupted Δmgd1/Δgcs1 and Δadh1/Δgcs1 supported the concept of an unbalanced redox state independent of the Glr1 activity for Δmgd1/Δadh1/Δgcs1. Our data demonstrate the reciprocal use of Eapx1 and Ccp1 in the absence of both methylglyoxal scavengers; that being pivotal for viability in non-filamentous budding yeast.

Comprehensive Characterization of Mutant Pichia stipitis Co-Fermenting Cellobiose and Xylose through Genomic and Transcriptomic Analyses

  • Dae-Hwan Kim;Hyo-Jin Choi;Yu Rim Lee;Soo-Jung Kim;Sangmin Lee;Won-Heong Lee
    • Journal of Microbiology and Biotechnology
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    • v.32 no.11
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    • pp.1485-1495
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    • 2022
  • The development of a yeast strain capable of fermenting mixed sugars efficiently is crucial for producing biofuels and value-added materials from cellulosic biomass. Previously, a mutant Pichia stipitis YN14 strain capable of co-fermenting xylose and cellobiose was developed through evolutionary engineering of the wild-type P. stipitis CBS6054 strain, which was incapable of co-fermenting xylose and cellobiose. In this study, through genomic and transcriptomic analyses, we sought to investigate the reasons for the improved sugar metabolic performance of the mutant YN14 strain in comparison with the parental CBS6054 strain. Unfortunately, comparative whole-genome sequencing (WGS) showed no mutation in any of the genes involved in the cellobiose metabolism between the two strains. However, comparative RNA sequencing (RNA-seq) revealed that the YN14 strain had 101.2 times and 5.9 times higher expression levels of HXT2.3 and BGL2 genes involved in cellobiose metabolism, and 6.9 times and 75.9 times lower expression levels of COX17 and SOD2.2 genes involved in respiration, respectively, compared with the CBS6054 strain. This may explain how the YN14 strain enhanced cellobiose metabolic performance and shifted the direction of cellobiose metabolic flux from respiration to fermentation in the presence of cellobiose compared with the CBS6054 strain.