• KSBB Journal
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• 제5권3호
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• pp.255-261
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• 1990

Astaxanthin을 생산하는 효모 Phaffia rhodozyma로부터 제조된 상보적 돌연변이 균주간의 세포융합을 통하여 astaxanthin을 대량 생산하는 균주를 얻고자 시도하였다. 이들의 원형질체 융합빈도는 $1.3$\times$10^-^5-6.0$\times$10^-^5$이었고 DNA함량, 핵염색, UV조사에 대한 생존력 비교 그리고 형질분리분석등으로 핵융합을 확인하였다. 융합체 중 F1은 야생형의 모 균주와 비교했을때 astaxanthin생성량이 약 3배 증가하였다.

• KSBB Journal
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• 제17권2호
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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.

• Toxicological Research
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• 제22권4호
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• pp.381-390
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• 2006

Astaxanthine is a pigment that belongs to the family of the xanthophylls, the oxygenated derivatives of carotenoids whose synthesis in plants derives from lycopene. Astaxanthine is also a carotenoid widely used in salmonid and crustacean aquaculture to provide the pink color characteristic of that. Recent study reported that astaxanthine has the role as a detoxicant against the free radicals. On our study, we estimated the genotoxicity in ICR mice and possibility as antioxidant reagents of mutant Phaffia rhodozyma strain over expressing the astaxanthine by gamma-lay and carophyll pink including astaxanthine in apoE knock out mice, respectively. In our study, we administered Phaffia rhodozyma (2 mg and 3 mg) and carophyll pink for 4 and 8 week. The clinical sign and mortality were not detected compared with control groups. In the mutant frequency of hprt gene and chromosome aberration in splenic cells, there was not detected abnormality. There was not critical change in hematological and serum biochemical test compared to control. In expression level of repair enzyme, increase of catalase were detected and increase of expression level of Nrf-2 was detected in Phaffia rhodozyma (3 mg) and carophyll pink in 8 week treated group. In GSH level, the group of treated with Phaffia rhodozyma (3 mg) showed the increase of the GSH. In conclusion, mutant Phaffia rhodozyma and caphyll pink may be applied to the effective food additives to reduce the free radical.

• 한국양식학회지
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• 제9권4호
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• pp.363-375
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• 1996

천연 astaxanthin 색소를 다량 함유한 phaffia 효모를 양식 어류에게 공급하였을 때, 이 색소가 어류의 성장과 소화, 흡수되는 정도를 조사하기 위하여 이스라엘 잉어($650{\pm}20$ g), 비단잉어($80{\pm}10$ g) 및 나일틸라피아($200{\pm}20$ g)의 사료에 phaffia 효모를 $10\%$ 첨가하여 10주간 공급한 다음 성장률, 체조성, 근육의 탄력도 및 근육 색과 체색 등의 변화를 측정하였다. Phaffia 효모 첨가 사료에 대한 효모 대조구로서 $10\%$의 맥주 효모 첨가 사료구를, 그리고 이들 두가지 반정제 효모 사료에 대한 성장 대조구로서 시판 상품 사료구를 설정하여 10주간 사육한 결과 이스라엘 잉어, 비단잉어 및 틸라피아의 성장에는 모든 사료 공급구에서 유의적인 차이가 없었다(P>0.05). 이들 실험어의 근육 내의 일반 성분은 실험 후에도 현저한 차이가 나타나지 않아(P>0.05), phaffia 효모나 맥주 효모 사료가 체성분 조성에는 영향이 없는 것으로 나타났다. 또한 횟감으로 이용되는 어종인 이스라엘 잉어와 틸라피아의 근육에 쫄깃한 정도를 나타내는 기계적 탄력도와 관능적 탄력도를 측정한 결과 모두 유의적인 차이는 나타나지 않았다(P>0.05). 실험어의 근육에 phaffia 효모에서 유래되는 붉은 색소의 침착은 이스라엘 잉어와 비단잉어 실험구에서는 근육내의 색소 분석과 관능적 측정 실험 모두에서 현저하게 높게 나타났지만(P<0.05), 틸라피아에서는 색소 분석과 관능적 측정 실험 모두에서 유의적인 차이가 나타나지 않았다(P>0.05).

• 한국동물학회:학술대회논문집
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• 한국동물학회 1994년도 한국생물과학협회 학술발표대회
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• pp.259-259
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• 1994

• 한국식품과학회지
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• 제31권3호
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• pp.720-726
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• 1999

고전압 펄스 전기장(PEF)가 P. rhodozyma 세포의 형태학적 변화와 carotenoid의 추출에 미치는 영향을 연구하였다. 전기장 세기 $10{\sim}50\;kV/cm$와 처리시간 $100{\sim}300\;{\mu}s$의 범위에서 세포를 PEF 처리했을 때 전기장의 세기와 처리시간이 증가함에 따라 세포의 팽창, 손상 정도와 세포내 물질이 유출되는 정도가 증가하였다. $10{\sim}50\;kV/cm$, 100 Hz의 exponential decay파로 세포현탁액을 $100\;{\mu}s$ 또는 $300\;{\mu}s$ 처리하였을 때 최대 전기장(50 kV/cm)에서 생균수가 각각 1.5 및 2.5 log 감소하였다. 50 kV/cm의 전기장에서 세포막에 형성되는 electroporation 정도는 98%에 달하였고, 이 때 세포의 회복률은 5% 미만으로 확인되었다. Phloxine B 색소로 세포를 염색했을 때 생균세포는 염색되지 않았으나, PEF 처리한 세포는 색소가 내부로 침투되어 적색으로 염색되었으므로 세포막이 손상된 것을 알 수 있었다. Carotenoid 색소가 P. rhodozyma 세포막의 지방체와 결합한 상태로 존재하기 때문에 고전압 PEF 처리에 의한 세포막 투과성 증진의 효과만으로는 색소 추출이 어려웠으나, 세포현탁액 조제시에 0.01% NaCl 용액 대신에 0.01% $CaCl_2$ 용액을 사용하는 경우에는 $10{\sim}20\;{\mu}g$의 색소 추출 증진 효과가 있었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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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.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.301-304
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• 2000

Fed-batch culture was designed to increase cell concentration and astaxanthin content by mutant AJ-6 of Phaffia rhodozyma. Fed-batch culture was performed in the continuous feeding with manual adjustment of flow rate to control glucose concentration. When the final glucose concentration was 100 g/L, the cell and astaxanthin were 38.3 g/L, 34.8 mg/L, respectively. Addition of ethanol(10 g/L), when glucose was depleted, the cell and astaxanthin concentration were 37.2 g/L and 45.6, respectively, 5 g/L of acetic acid supplied, 40.6 g/L, 43.9 mg/L were obtained. Ethanol and acetic acid enhanced the astaxanthin content act as precursor of carotenoid synthesis.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.314.1-314
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• 1995

• Journal of Microbiology and Biotechnology
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• 제8권6호
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• pp.650-655
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• 1998

The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.