• Journal of Applied Biological Chemistry
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• v.56 no.3
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• pp.171-177
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• 2013

Efficient extraction of total proteins from soil microorganisms is tedious because of small quantity. In this regard, an improved method for extraction of whole cell proteins is developed from soil microorganisms, Saccharomyces cerevisiae and Pichia pastoris. of which the cell wall are very strong. Pretreatment with NH4OH prior to the final extraction using NaOH/SDS was tried under the basis that ammonium ion was possible to enhance the permeability and/or to weaken the yeast cell walls. The pre-treatment of yeast cells with NH4OH drastically enhanced the protein extraction when it was compared with control (without NH4OH pre-treatment). At the pre-treatment of 0.04 N NH4OH at pH 9.0, about 3 fold of proteins was obtained from p. pastoris. Ammonium hydroxide appears to penetrate into the yeast cell walls more readily at basic pH. The effect of NH4OH pretreatment was pH dependent. The methods developed in this experiment might be applicable for an effective extraction of yeast proteins for the purpose of biochemical studies, especially proteomic analysis.

• Mycobiology
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• v.38 no.1
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• pp.70-73
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• 2010

Temperature-sensitive yeast mutants were used to screen for cell cycle-related genes from Pleurotus eryngii genomic DNA. A mushroom genomic DNA library was established and each gene was screened for the ability to rescue seven Saccharomyces cerevisiae temperature-sensitive strains. Hundreds of yeast transformants were selected at restrictive temperatures over $30^{\circ}C$. Plasmids from the transformants that survived were isolated and transformed back into their host strains. The temperature sensitivity of the resulting transformants was tested from $30^{\circ}C$ to $37^{\circ}C$. Ten DNA fragments from P. eryngii were able to rescue yeast temperature-sensitive strains, and their DNA sequences were determined.

• Proceedings of the Korean Institute of Resources Recycling Conference
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• 2001.05b
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• pp.121-125
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• 2001

For the probiotic feed production, aerobic liquid fermentation of pulverized food wastes was attempted with a yeast Kluyveromyces marxianus. After grinding finely, optimal fermentation conditions of the substrate was investigated by shaking culture. The most active growth of the yeast was shown at solid content of 10%. The proper addition of urea(0.5g/l), o-phosphate(0.4g/l), molasses(4g/l), and yeast extract (1g/1) increased cell growth rate and viable cell count. For optimizing, the nutrients were all added to substrate and fermentation was carried in 2 litre jar fermenter. For the stimulation of hydrolyzing enzyme excretion, mixed culture with Aspersillus oryzae was also conducted. In 12 hours of fermentation, viable cell count of the yeast Kluyveromyces marxianus amounted to the number of 1.4 $\times$10$^{10}$ /1 in the culture medium.

• Journal of Microbiology and Biotechnology
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• v.32 no.11
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• pp.1373-1381
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• 2022

Fructan is a polysaccharide composed of fructose and can be classified into several types, such as inulin, levan, and fructo-oligosaccharides, based on their linkage patterns and degree of polymerization. Owing to its structural and functional diversity, fructan has been used in various fields including prebiotics, foods and beverages, cosmetics, and pharmaceutical applications. With increasing interest in fructans, efficient and straightforward production methods have been explored. Since the 1990s, yeast cells have been employed as producers of recombinant enzymes for enzymatic conversion of fructans including fructosyltransferases derived from various microbes and plants. More recently, yeast cell factories are highlighted as efficient workhorses for fructan production by direct fermentation. In this review, recent advances and strategies for fructan biosynthesis by yeast cell factories are discussed.

• Applied Chemistry for Engineering
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• v.29 no.4
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• pp.419-424
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• 2018

In this work, effect of the culture medium composition on the fermentation process of Clostridium ljungdahlii, which is acetogenic bacteria to product ethanol from synthesis gas, was examined to improve the microbial growth and ethanol production. Components of the culture medium such as yeast extract, fructose, $NH_4Cl$, and $K_2HPO_4$ were selected as influence factors for the cell growth and ethanol production. As the concentration of yeast extract increased, both of the cell growth and ethanol production increased. And the ethanol productivity was the highest at an yeast extract of 0.05 g/L, which is lower than that of base medium. As the concentration of fructose increased, the cell growth increased, but the ethanol production decreased when the concentration of fructose was higher than that of base medium (5 g/L). In an experiment with the yeast extract of 5 g/L, produced ethanol concentration was the highest (0.297 g/L) when fructose concentration was 5 g/L, however, the specific ethanol productivity was higher (0.281 g/g DCW) when the fructose was not added due to very low cell mass. The cell growth and ethanol production were not significantly influenced by $NH_4Cl$ concentration, however the growth inhibition was observed at a 30 g/L of $NH_4Cl$. When the concentration of $K_2HPO_4$ increased, both of the cell growth and ethanol production increased. In experiments with $NH_4Cl$ and $K_2HPO_4$, specific ethanol productivities were higher when the low concentration of yeast extract was used.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.77-77
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• 2003

Gene expression in a cell is regulated by mutual activations or repressions between genes. Identifying the gene regulation network will be one of the most important research topics in the post genomic era. We propose a linear dynamic model of gene regulation for the yeast cell cycle. A small gene network consisting of about 40 genes is reconstructed from the analysis of micro-array gene expression data of yeast S. cerevisiae published by P. Spellman et al. We show that the network construction is consistent with the result of the hierarchical cluster analysis.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.195-198
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• 2006

Lethal toxin is a critical virulence factor of anthrax. It is composed two protein: protective antigen (PA) and lethal factor (LF). PA binds to specific cell surface receptors and, forms a membrane channel that mediates entry of LF into the cell. LF is a zinc-dependent metalloprotease, which cleaves MKKs [MAPK (mitogen-activated protein kinase) kinases] at peptide bonds very close to their N-termini. In this study, we suggest application of cell-based assays in the early phase of drug discovery, with a particular focus on the use of yeast cells. We constructed MEK1 expression system in yeast to determine LF activity and approached cell-based assay system to screen inhibitors, in which the results covering the construction of LF-substrate in yeast expression vector, expression, and LF-mediated proteolysis of substrate were described. These results could provided the basic steps in design of cell-based assay system with the high efficiency, rapidly and easy way to screening of inhibitors.

• Food Science and Biotechnology
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• v.15 no.4
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• pp.506-510
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• 2006

To be used as a source of astaxanthin by animals, the red yeast Xanthophyllomyces dendrorhous needs to be dried and the cell wall ruptured. Spray-drying and flat-roller milling successfully prepared the yeast as a feed additive with little loss of astaxanthin. Spray-drying successfully dried the yeast with negligible decomposition of astaxanthin compared to drum-drying. By repeated milling with a flat-roller mill, astaxanthin extracted with ethanol increased from 0.01 to 1.31 mg astaxanthin/g yeast. This method did not decompose astaxanthin in contrast to chemical digestion of the cell wall. Flat-roller milling effectively flattened and cracked the dried cells. Astaxanthin in yeast prepared by spray-drying and flat-roller milling was well absorbed by animals. Specifically, when spray-dried and milled yeast was supplied in the feed (40 mg astaxanthin/kg feed), astaxanthin was successfully absorbed (1,500 ng/mL blood and 1,100 ng/g skin) by laying hens.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.4
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• pp.288-305
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• 2000

A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects-the basics of yeast flocculation, the development of "new" flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous $\beta$-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculating bioreactors and discussing potential new uses of these systems.e systems.