• Journal of Microbiology and Biotechnology
• /
• v.8 no.5
• /
• pp.501-508
• /
• 1998

An alkalophilic mi.croorganism (strain YB380), which produces yeast cell wall hydrolase extracellulary, was isolated from Korean soil. The rod-shaped cells were 0.3~0.4 by 2~4${\mu}{\textrm}{m}$ long, motile, aerobic, gram-positive, and spore-forming. The color of the colony was light yellow. The temperature range for growth at pH 9.0 was 25 to $45{\circ}C, with optimum growth at $35{\circ}C. The pH range for growth at $35{\circ}C was 8 to 11 with an optimum pH of 9.0. Therefore, the strain YB380 is an obligate alkalophile. The 16S rRNA of strain YB380 has a 99% sequence similarity with that of Bacillus alcalophilus. On the basis of physiological properties, cell wall fatty acid composition, and phylogenetic analysis, we propose that the isolated strain is Bacillus alcalophilus. The yeast cell wall hydrolase from Bacillus alcalophilus subsp. YB380 has been purified and partially characterized. The molecular weight was estimated to be 27,000 daltons with an optimum temperature and pH of $60{\circ}C and 9.0, respectively. The N-terminal amino acid sequence of the enzyme was analyzed as Gln- Thr- Val- Pro- Trp- Gly- Ile- Asn- Arg- Val.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.4
• /
• pp.767-774
• /
• 2010

In this study, the problems of high caloric content, increased maturation time, and off-flavors in commercial beer manufacture arising from residual sugar, diacetyl, and acetaldehyde levels were addressed. A recombinant industrial brewing yeast strain (TQ1) was generated from T1 [Lipomyces starkeyi dextranase gene (LSD1) introduced, ${\alpha}$-acetohydroxyacid synthase gene (ILV2) disrupted] by introducing Saccharomyces cerevisiae glucoamylase (SGA1) and a strong promoter (PGK1), while disrupting the gene coding alcohol dehydrogenase (ADH2). The highest glucoamylase activity for TQ1 was 93.26 U/ml compared with host strain T1 (12.36 U/ml) and wild-type industrial yeast strain YSF5 (10.39 U/ml), respectively. European Brewery Convention (EBC) tube fermentation tests comparing the fermentation broths of TQ1 with T1 and YSF5 showed that the real extracts were reduced by 15.79% and 22.47%; the main residual maltotriose concentrations were reduced by 13.75% and 18.82%; the caloric contents were reduced by 27.18 and 35.39 calories per 12 oz. Owing to the disruption of the ADH2 gene in TQ1, the off-flavor acetaldehyde concentrations in the fermentation broth were 9.43% and 13.28%, respectively, lower than that of T1 and YSF5. No heterologous DNA sequences or drug resistance genes were introduced into TQ1. Hence, the gene manipulations in this work properly solved the addressed problems in commercial beer manufacture.

• Food Science and Biotechnology
• /
• v.17 no.3
• /
• pp.558-563
• /
• 2008

To prevent weakly flocculating characters of bottom brewing yeast during first fermentation, various technical investigations were carried out using strain of Saccharomyces cerevisiae. It appeared that the propagation at $10^{\circ}C$ promoted the molecular structure and biochemical composition of cell wall in favor of flocculation. The yeast grown at $20^{\circ}C$ by addition of zinc ion also had a stimulating effect on flocculation behavior during first fermentation cycle. The zinc ion did not influence directly on the changes of cell wall in favor of stronger flocculence. The increased fermentation activity of yeast due to addition zinc ion was rather responsible for the intensified flocculation capacity. It was concluded that the weakly flocculating characters of bottom brewing yeast during first fermentation can be solved by using yeast propagated at $10^{\circ}C$ or by means of yeast by addition of zinc ion during propagation.

• Journal of Life Science
• /
• v.16 no.3 s.76
• /
• pp.454-458
• /
• 2006

HSF1 is the heat shock transcription factor in Saccharomyces cerevisiae. S. cerevisiae KNU5377 can ferment at high temperature such as $40^{\b{o}}C$. We have been the subjects of intense study because Hsf1p mediates gene expression not only to heat shock, but to a variety of cellular and environmental stress challenges. Basing these facts, we firstly tried to construct the hsf1 gene-deleted mutant. PCR-method for fast production of gene disruption cassette was introduced in a thermotolerant yeast S. cerevisiae KNU5377, which allowed the addition of short flanking homology region as short as 45 bp suffice to mediate homologous recombination to kanMX module. Such a cassette is composed of linking genomic DNA of target gene to the selectable marker kanMX4 that confers geneticin (G418) resistance in yeast. That module is extensively used for PCR-based gene replacement of target gene in the laboratory strains. We describe here the generation of hsf1 gene disruption construction using PCR product of selectable marker with primers that provide homology to the hsf1 gene following separation of haploid strain in wild type yeast S. cerevisiae KNU5377. Yeast deletion overview containing replace cassette module, deletion mutant construction and strain confirmation in this study used Saccharomyces Genome Deletion Project (http:://www-sequence.standard.edu/group/yeast_deletion_project). This mutant by genetic manipulation of wild type yeast KNU5377 strain will provide a good system for analyzing the research of the molecular biology underlying their physiology and metabolic process under fermentation and improvement of their fermentative properties.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.11
• /
• pp.1557-1567
• /
• 2012

Glutathione reductase (GR, E.C. 1.6.4.2) is an important enzyme that reduces glutathione disulfide (GSSG) to a sulfydryl form (GSH) in the presence of an NADPH-dependent system. This is a critical antioxidant mechanism. Owing to the significance of GR, this enzyme has been examined in a number of animals, plants, and microbes. We performed a study to evaluate the molecular properties of GR (OsGR) from rice (Oryza sativa). To determine whether heterologous expression of OsGR can reduce the deleterious effects of unfavorable abiotic conditions, we constructed a transgenic Saccharomyces cerevisiae strain expressing the GR gene cloned into the yeast expression vector p426GPD. OsGR expression was confirmed by a semiquantitative reverse transcriptase polymerase chain reaction (semiquantitative RT-PCR) assay, Western-blotting, and a test for enzyme activity. OsGR expression increased the ability of the yeast cells to adapt and recover from $H_2O_2$-induced oxidative stress and various stimuli including heat shock and exposure to menadione, heavy metals (iron, zinc, copper, and cadmium), sodium dodecyl sulfate (SDS), ethanol, and sulfuric acid. However, augmented OsGR expression did not affect the yeast fermentation capacity owing to reduction of OsGR by multiple factors produced during the fermentation process. These results suggest that ectopic OsGR expression conferred acquired tolerance by improving cellular homeostasis and resistance against different stresses in the genetically modified yeast strain, but did not affect fermentation ability.

• Microbiology and Biotechnology Letters
• /
• v.23 no.1
• /
• pp.12-16
• /
• 1995

Medium components for lactic acid production were optimized with a strain of Lactobacillus sp., isolated by our Lab. Nitrogen source was the key component and manganese ion was also important for lactic acid production in this strain. Optimal concentration of manganese ion was 0.03 g/l as MnSO$_{4}$ 4 - 5 H$_{2}$O base. Other mineral elements, however, had little effect on it. Among the nitrogen sources we examined, yeast extract showed the highest productivity. Yeast extract, the exellent but very expensive medium component, could be partially replaced by soytone until 60% dry base with higher productivity, or by tryptone enforced with vitamines and nucleic acids. In order to replace yeast extract completely, we examined several inexpensive nitrogen sources and their enzymatic hydrolyzates. The hydrolyzate of vital wheat gluten was the best of them.

• Microbiology and Biotechnology Letters
• /
• v.23 no.6
• /
• pp.723-729
• /
• 1995

A flocculating sugar tolerant yeast strain was isolated from fermenting Takju. This strain was identified as Saccharomyces cerevisiae CA-1 according to the Lodder's yeast taxonomic studies. The isolated yeast could grow in 50% glucose and in 7% ethanol in the YPD medium. It's optimal growth temperature, initial pH, shaking rate and initial glucose concentration for ethanol fermentation showed 35$\circ$C, 4.5, 150 rpm, 15%, respectively. Ethanol concentration was 63 g/l in 20% glucose after 24 hours, fermentation yield was 0.49 g-ethanol/g-glucose in 10% glucose after 24 hours and ethanol productivity was 3.09 g/l$\cdot $h in 10% glucose after 12 hours in batch fermentation. Repeated batch fermentation was possible for over 50 days and ethanol yield, ethanol productivity and substrate conversion rate were 0.39-0.50 g/g, 1.63-2.08 g/l$\cdot $h and more than 99%, respectively during these periods.

• Korean Journal of Microbiology
• /
• v.19 no.4
• /
• pp.174-178
• /
• 1981

Psychrophilic yeast strains, HJ3011, HJ3023, and HJ3021, which were isolated from the Han River during winter season of December 1980 to February 1981 were identified as Candida sp., Candida nivalis, and Rhodotorula rubra respectively. The effect of temperature on growth was carried out in this study. Growth rate constants of strains HJ3011 and HJ3023 were the highest value at $15^{\circ}C\;and\;16{\sim}20^{\circ}C$ respectively, and both growth rates diminished gradually above $20^{\circ}C$ and were decreased to zero at $25^{\circ}C$. According to the results obtained in this experiment, above two strains of yeast were confirmed as psychrophile. On the other hand since the strain of HJ3021 exhibited the highest growth rate constant at $15^{\circ}C$ and could slightly grow above $25^{\circ}C$, this strain was defined as psychrotroph.

• Mycobiology
• /
• v.49 no.5
• /
• pp.521-526
• /
• 2021

Three isolates belonging to the ascomycetous genus Zygotorulaspora were obtained from the fruits of Cornus officinalis and Smilax china, and flowers of Dendranthema zawadskii var. latilobum in Gongju-si, Korea. Phylogenetic Analyses of the LSU D1/D2 domain and ITS region sequences supported the recognition of two new species: Zygotorulaspora cornina sp. nov. (type strain NIBRFGC000500475 = KACC93346P^PP) and Zygotorulaspora smilacis sp. nov. (type strain NIBRFGC000500476 = KACC93347P^PP). The two novel species revealed no growth on D-Galactose, unlike the other six species in the genus Zygotorulaspora. They are distinguished from each other by their phylogenetic differences and phenotypic characteristics such as assimilation of xylitol, 5-keto-D-gluconate, and ethanol. All species in the genus Zygotorulaspora including the two novel species have phenotypic traits of genus Zygotorulaspora: asci are persistent, sucrose and raffinose are assimilated, and m-inositol is not required for growth, and they are mainly associated with plants.

• The Korean Journal of Mycology
• /
• v.51 no.4
• /
• pp.373-381
• /
• 2023

This study aimed to isolate and characterize wild yeast strains from the vineyard soil in Korea. Twenty yeast strains were isolated from vineyard soil in Gimpo-si, Gyeonggi-do, Korea, where Vitis labrusca cv. Campbell Early is grown. Eighteen strains were previously recorded in Korea. The remaining two, Cyberlindnera mrakii VG-21-10C and Starmerella bacillaris GR9 were not previously recorded in Korea. The mycological characteristics of VG-21-10C and GR9 were investigated. Both were oval-shaped, convex, and smooth. However, differences were evident in colony color and carbon assimilation activities. Strain VG-21-10C is white-colored and assimilates glucose, glycerol, D-xylose, D-cellobiose, D-saccharose, and D-raffinose as carbon sources. Strain GR9 is cream-colored and assimilates glucose, D-saccharose, and D-raffinose as carbon sources.