• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.463-470
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• 1996

The red yeast Phaffia rhodozyma, which contains astaxanthin(3, 3'-dihydroxy-$\beta$, $\beta$-carotene-4, 4'-dione) as its primary carotenoid, was tested as a dietary pigment source for egg yolks of laying hens. When the yeast was fed to laying hens at several concentrations, the intensity of redness in egg yolks was dependent on the yeast concentration in the feed and the deposition period. Addition of P rhodozyma in feed did not cause any visible adverse effect on laying hens.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.103-109
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• 1996

Mn(II)+succinate decreased the carotenoid formation of the yeast Phaffia rhodozyma, probably by scavenging $O_2$. When duroquinone (DQ), an internal and external $O_2$ generator, was added to medium, P. rhodozyma produced more amount of carotenoids. The increased carotenoid production was destroyed by oxygen radical (OR) scavengers, ascorbate+Cu(II) and dimethylsulfoxide. When sub-lethal concentrations of $H_2O_2$ , an external OR source, and antimycin, an internal OR inducer, were used, the effect of $H_2O_2$ on carotenoid formation and composition was less significant than that of antimycin. Addition of superoxide dismutase, an external OR remover, rescued cells from death caused by the high concentration of DO. In this condition, the yeast culture showed an increase in carotenoid content. Addition of DQ into P. rhodozyma culture in the stationary phase did not increase carotenoid production. Therefore, carotenoid formation was stimulated by internal ORs in the growing yeast. It was probably due to release of catabolite repression on carotenogenesis in the yeast. Aeration was important for carotenoid production but was not as effective as the internal OR producer, DQ.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.110-115
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• 1996

Yellow mutants of the astaxanthin producing red yeast Phaffia rhodozyma were obtained by nitrosoguanidine mutagenesis. The carotenoid composition of the yelow mutants, Yan-1 and Ny-1, was mainly $\beta$ -carotene (> 95$%$) and torulene (< 5$$). Therefore, the yellow mutants are carotene oxygenation deficient mutants (CODMs). CODMs produced decreased quantities of carotenoids compared to their red parents and this indicated that carotene may regulate its synthesis. CODMs, Yan-1 and Ny-4, on plates containing 50 $\mu$ M antimycin, showed highly pigmented vertical papillae. Antimycin-induced mutants purified from the papillae showed increases in carotenoid content (up to 1 mg $\beta$-carotene/g yeast). CODMs, Yan-1 and Ay-1, were more sensitive to antimycin than red strains, Ant-1 and 67-385. This was probably due to lower antioxidant activity of $\beta$-carotene than that of astaxanthin. Light increased torulene and light+antimycin further increased the torulene. Yan-1 and Ny-4 could grow with succinate, though their red parents, Ant-1 and Anf-1p, could not. However, antimycin induced mutation of Yan-1 or Ny-4 destroyed the ability to grow with succinate.

• Journal of Microbiology and Biotechnology
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• v.2 no.1
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• pp.46-49
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• 1992

The availability of Phaffia rhodozyma as an astaxanthin sources in the aquaculture industry is limited because of the low carotenoid content of natural isolate. In this study, we have used the protoplast fusion technique to construct cell hybrids with an increased content of astaxanthin from P. rhodozyma. Cell hybrids (F307 and F406) obtained were very stable and produced considerably more astaxanthin (> 1 mg/g yeast) than the wild parent. Karyogamy was confirmed by the isolation of recombinants after mitotic segregation of parental auxotrophic genetic markers, the increased amount of chromosomal DNA/cell and the presence of single nucleus/cell.

• KSBB Journal
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• v.17 no.2
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• pp.176-181
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• 2002

Astaxanthin (3,3＇-dihydroxy-${\beta}$, ${\beta}$-carotene-4-4＇-dione), a natural pigment of pink to red color, is widely distributed in nature particularly in the skin layer of salmonoids and the crust of shrimp, lobster, etc. Recently, it was produced from the yeast culture of Phaffia rhodozyma. Because of its high thermal stability and antioxidant functionality, its applications can be extended into food, cosmetics, and pharmaceutical ingredient beyond the traditional feed additive. Because of its very high lipophilicity, astaxanthin has been extracted traditionally by strong organic solvents such as chloroform, petroleum ether, acetone, etc. In this study, we developed a surfactant-based solubillization system for astaxanthin, and used it to extract astaxanthin from disrupted yeast cells. Among Tween 20, Triton X-100 and SDS, Tween 20 was identified as the most suitable surfactant in terms of extraction capacity and safety. The ethylene oxide group of Tween 20 was identified as the most significant factor to increase the HLB value that determined the extraction capacity. The effects of micelle formation condition, such as the molar ratio of astaxanthin and Tween 20, pH, and ionic strength were also investigated. pH and ionic strength showed no significant effects. The optimal molar ratio between astaxanthin and Tween 20 was 1 : 12. Antioxidant activity of astaxanthin was higher than ${\beta}$-carotene and ${\alpha}$-tocopherol. Astaxanthin in the crude extract from the yeast cell was more resistant to air and/or light degradation than pure astaxanthin, probably because of the presence of other carotenoids and lipids.

• Journal of Aquaculture
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• v.9 no.4
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• pp.363-375
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• 1996

The effects of the yeast, Phaffia rhodozyme in the diet on growth, body composition, muscle elasticity and pigmentation of Israeli strain of common carp, colored carp and Nile tilapia were investigated. Ten percent of the yeast was added to semi purified diet as an experimental feed 1. Ten percent of brewers yeast in the semi purified diet (experimental feed 2) was tested for comparing the growth performance between two semi purified diets. A commercial diet was also used for the check of growth rate of the semi purified diets. All experimental fish were fed for 10 weeks. The weight gains among the experimental fish were not significantly different (P>0.05). There were not significantly different in body composition, muscle elasticity among the the fishes fed three experimental diets. There were significant differences (P<0.05) of pigment deposition in the muscle of Israeli strain of common can and on the skin of colored carp between treated and non treated group. But there were no differences of pigment deposition in flesh and skin of tilapia among the three diets.

• Korean Journal of Food Science and Technology
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• v.31 no.3
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• pp.720-726
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• 1999

High voltage pulsed electric fields (PEF) technology is a non-thermal technique which is applicable to extract useful components froms biological materials. This research suggested the possibility for extracting carotenoid pigments from Phaffia rhodozyma by PEF treatments. The yeast cell suspensions were treated with high voltage pulses in a recycled PEF treatment chamber which consists a pair of thin plates of stainless steel adhering to a small chamber with approximately $1{\sim}4\;mm$ gap. A 2.5 log reduction in survivability and more than 98% of electropermeabilization of the yeast cells could be achieved by PEF treatment for $300\;{\mu}s$ with an electric field of 30 kV/cm and pulse duration of $1\;{\mu}s$. When the yeast cell suspended in 0.01% NaCl solution were treated with PEF under various conditions, carotenoid pigments were not extracted. However, the PEF treatment of the yeast cell suspensions in 0.01% $CaCl_2$ solution, have positive effects on the extraction of carotenoid pigments ($27.3\;{\mu}g/g$ of dried yeast).

• KSBB Journal
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• v.5 no.3
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• pp.255-261
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• 1990

Cell fusion between complementary mutants isolated from astaxanthin-producing yeast, Phaffia rhodozyma, was carried out to obtain astaxanthin-overproducing strains by protoplast fusion technique. The frequency of protoplast fusion was ranged from 2.3$\times$10-5 to 6.0$\times$10-5, and nuclear fusion in the cells of hybrids was demonstrated by several techniques such as isolation of recombinants after mitotic segregation of parental genetic markers, estimation of DNA content, direct observation of nuclei with nuclear staining, and comparison of survival rate to UV exposure. One of several hybrids, Fl, showed approximately 3-fold increase in astaxanthin content when compared with wild parent.

• The Korean Journal of Mycology
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• v.21 no.1
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• pp.9-15
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• 1993

ABSTRACT: The cell fusants were constructed from complementary auxotrophic mutants of Phffia rhodozyma. The nuclear fusion of the fusants was demonstrated by several techniques including comparision of cell volume, estimation of DNA content and nuclear staining. The cell fusants were very stable for succeeding transfer culture on complex medium for more than one year. Malt extract (1%, w/v) and abscisic acid(1 mM) increased the carotenoid formation whereas gibberellic acid(5mM) and riboflavin(0.1 mM) decreased the corresponding content.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.271-274
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• 2000

A study was carried out to select a nitrogen source and the optimize the C/N ratio for the maximum cell growth of Phaffia rhodozyma in fed-batch culture. The yeast extract was the best organic nitrogen source. In the batch culture experiments, the highest cell yield was obtained 0.575 g-cell/g-glucose from 10 g/L and 10 g/L yeast extract. In the fed-batch experiments, the maximum cell concentration was obtained 33.1 g/L from the C/N ratio of 2:1 while the astaxanthin concentration of cell was Increased by increasing the C/N ratio, of feed medium.