• 한국미생물·생명공학회지
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• 제18권3호
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• pp.233-238
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• 1990

The mechanism of intracellular accumulation of cadmium in a cadmium-ion tolerant yeast, Hansenula ammala B-7, which is an extreme cadmium tolerant strain and has the ability to take up a large amount of cadmium was investigated. The amounts of cadmium taken up by the scalded yeast cells were 2 to 3 times more than the value of the living cells. The living Hansenula anomala B-7 cells adsorbed 74% of cadmium taken up onto the other layer of the cells and 26% of it accumulated inside the cells. But the scalded cells adsorbed 98.3% of cadmium taken up and accumulated 1.7% of it inside the cells. A cadmium uptake and its accumulation were accelerated up to 162.3% and 275.4% by Triton X-100 in the living cells, respectively. Whereas in the scalded cell cadmium uptake was not affected by Triton X-100. Furthermore the cadmium uptake and its accumulation were strongly inhibited by metabolic inhibitors like 2,4-dinitrophenol, sodium azide and potassium cyanide in the living cells, but in the scalded cells cadmium uptake was not affected by metabolic inhibitors. These results suggested that the intracellular accumulation of cadmium by the cadmium-tolerant Hansenula anomala B-7 cells was apparently dependent of biological activity, and also gave evidence of the existance of energy-dependent system.

• 한국환경농학회지
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• 제33권2호
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• pp.129-133
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• 2014

BACKGROUND: This study was performed for selecting yeast mutants with a high tolerance for targeted metals, and determining whether yeasts strains tolerant to multiple heavy metals could be induced by sequential adaptations. METHODS AND RESULTS: A mutant yeast strain tolerant to the heavy metals cadmium (Cd), copper (Cu), nickel (Ni), and zinc (Zn) was selected by sequential elevated exposures to each metal with intermittent mutant isolation steps. A Cd-tolerant mutant was isolated by growing yeast cells in media containing $CdCl_2$ concentrations that were gradually increased to 1 mM. Then the Cd-tolerant mutant was gradually exposed to increasing levels of $CuCl_2$ in growth media until a concentration of 7 mM was reached, thus generating a strain tolerant to both Cd and Cu. In the subsequent steps, this mutant was exposed to $NiCl_2$ (up to 8 mM), and a resultant isolate was further exposed to $ZnCl_2$ (up to 60 mM), allowing the derivation of a yeast mutant that was simultaneously tolerant to Cd, Cu, Ni, and Zn. CONCLUSION: This method of inducing tolerance to multiple targeted heavy metals in yeast will be useful in the bioremediation of heavy metals.

• Journal of Microbiology and Biotechnology
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• 제9권1호
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• pp.70-77
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• 1999

An extremely cadmium-tolerant budding yeast, Hansenula anomala B-7 underwent a morphological switch in response to either heat shock treatment or cadmium stress, respectively. It exhibited a morphological transition from a unicellular yeast form to a pseudohyphae-like coagulation when subjected to prolonged heat shock treatment. In contrast, the yeast cells showed an irregularity in surface morphology when given thermal stress for a short time. Patterns of proteins expressed in the pseudohyphae-like cells demonstrated that several proteins were overexpressed while others were underexpressed in comparison with those prepared from the cells in the yeast form. It was a striking feature, however, that nearly 40％ of the proteins extracted from the cells in the pseudohyphae form appeared to be composed of a single polypeptide. This polypeptide was apparently overexpressed during the pseudohyphae phase and its molecular weight was estimated to be 58 kDa according to SDS-PAGE analysis. However, a significant level of the protein was not observed in the cells before transition to pseudohyphae. The architecture of the cell shape was also damaged when incubated in a medium containing more than 1,000 ppm (8.9mM) of cadmium ions, although able to proliferate at a slow rate. However, the irregularity in the cell morphology exerted either by the brief heat shock treatment or by the cadmium stress with the high concentrations of the metal ions was not repaired, even though the damaged cells were allowed to grow for sufficient time in fresh, cadmium-free medium.

• 한국미생물·생명공학회지
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• 제24권3호
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• pp.268-273
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• 1996

An extremely cadmium tolerant yeast, Hansenula anomala B-7 used to determine the modification of the intracellular enzyme activities by cadmium ion. The activities of alcohol dehydrogenase, phosphofructokinase, and cytidine deaminase were decreased up to 90%, 40%, and 86% compa- red with the control by 1 mM cadmium nitrate respectively, but the activities of malate dehydrogenase, 6- phosphogluconate dehydrogenase, cytochrome c oxidase, and alkaline phosphatase were increased up to 440%, 136%, 260% and 155% compared with the control by 1 mM cadmium nitrate respectively. These results show that the activities of the enzymes participating in Embden-Mayerhof pathway (e.g. anaerobic metabolism) were reduced by cadmium, but those involved in hexose monophosphate pathway and tricarboxylic acid cycle (e.g. aerobic metabolism) were stimulated in contrast. It has been suggested that the diminished activity of cytidine deaminase in pyrimidine nucleotide dissimilation occured due to the inhibited nucleotide dissimilation by cadmium ion; the enhanced activity of cytochrome c oxidase was specifically required in order to oxidize a raised amount of NADH and NADPH due to the increased aerobic metabolism.

• 한국환경보건학회지
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• 제22권2호
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• pp.104-113
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• 1996

Cadmium uptake by growing and nongrowing (intact) cells of a chdmium-tolerant yeast Hansenula anomala B-7 in the presence of surfactants was studied. In growing cultures the addition of Triton X-100 or Tween 80 increased cadmium uptake by about 30% with no inhibition of cell growth, and in intact cells Triton X-100 increased cadmium accumulation by about 80% compared to surfactant-free controls. Considering balance between increased uptake and pollution, the addition of 0.1% Triton X-100 was preferable. By the mixed addition with defoamer silicone, during growth of cells Tween 80 or Triton X-100 enhanced uptake efficiency of cadmium compared to its single addition, whereas in intact biomass each of surfactants tested had no significant effect on cadmium uptake. The uptake of cadmium was observed to rise sharply to a maximum and then declined with increasing pH, and maximum accumulation of cadmium by growing and intact cells occurred at the pH of 6.0 and 7.0, respectively. A significant increase in cadmium uptake occurred with shaking culture. Cadmium uptake by growing and intact cells was almost completed during the culture time of 72 or 24 hrs, respectively. Scalded cells sorbed much more cadmium-ion than living cells.

• 미생물학회지
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• 제29권6호
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• pp.397-401
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• 1991

Yeast-form cells of cadmium ion-tolerant Hansenula anomala B-7 were changed to mycelial cells in medium containing more than $400\mu$g/ml of cadmium. Moreover, the mycelial cells were exchanged into clumped cells in a medium containing more than $1,000\mu$g/ml of cadmium. Optimal conditions of mycelial cell formation were achieved in the presence of .$1,000\mu$g/ml of cadmium with shaking cultivation for 7 days. Glucan and mannan contents of the yeast cell wall frown with $1,000\mu$g/ml of cadmium decreased by 10% compared with those grown without cadmium. However, protein and lipid contents increased about 20% respectively. By cadmium, no significant findings in specific amino acid contents were discovered.

• 미생물학회지
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• 제24권1호
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• pp.57-61
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• 1986

폐수중에 함유된 카드뮴을 미생물학적인 방법으로 정화하기 위한 기초 연구로서 아연 광산에서 고도 카드뮴 내성 효모 B-7을 분리하여 동정한 바 Hansenula anomala B-7 또는 그 유연끌으로 밝혀졌다. 이 효모는 농도 구배 한천 명판법에 의하여 $2,700{\mu}g/ml$의 카드뮴 농도에서도 내성을 나타내므로 고도 내성균으로 분류되었고 $1,000{\mu}g/ml$의 고농도 카드뮴 함유 액체 배지에서도 생육이 가능함을 확인하였다.

• 미생물학회지
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• 제25권2호
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• pp.110-116
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• 1987

고도 카드뮴 내성효모 Hansenula anomala B-7의 균체내에 다량의 카드뮴을 축적시키기 위하여 배양시 계면활성제를 첨가하여 배양하는 방법을 검토했다. 계면활성제 중 비이온계인 Triton X-100에 의하여 약 40%의 카드뮴의 축적효과를 나타내며 유효농도는 0.1-0.2%였다. 균체내 카드뮴 축적의 최적조건은 다음과 같다. 정치배양보다 진탕배양이 효과적이었으며, 48시간 진탕배양으로 카드뮴의 균체내 축적이 최대에 도달했으며 배지의 초기 pH는 6.0이었으며, 배양온도는 $40^{\circ}C$가 양호했다.

• 한국미생물·생명공학회지
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• 제17권1호
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• pp.29-34
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• 1989

고도 카드뮴 내성 효모, Hansenula anomala B-7 의 intact cell에 의한 카드뮴의 세포내 축적에 미치는 계면활성제의 영향를 검토하고, Triton X-100의 존재하에서의 카드뮴 축적에 미치는 제인자 등을 검토했다. 카드뮴의 축적은 0.1% Triton X-100과 Aerosol OT에 의하여 약 40% 이상 증가되었다. 카드뮴 이온에 대한 Km값은 0.247mM로 계산되었다. 카드뮴의 축적 최적 pH는 pH7.0에서 pH10.0 사이였으며, 최적온도는 4$0^{\circ}C$였다. 카드뮴의 최적 축적은 정치보다 진탕하므로 약 3배 증가되었으며, 진탕속도는 영향을 미치지 않았다. 카드뮴과 아연 이온을 공존시키므로 카드뮴의 축적이 아연 이온에 의하여 저해되었다.

• Journal of Microbiology and Biotechnology
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• 제4권1호
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• pp.30-35
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• 1994

A varient strain of budding yeast, Hansenula anomala B-7 which had been identified to be highly resistant to cadmium ions, were observed by transmission electron microscopy. It was shown that the cells accumulated excess amounts of cadmium ions throughout inside the cell rather than on the cell surface. The cell growth in response to cadmium or heat shock stress has also been investigated. It was observed that the cells precultured in the presence of 500 $\mu$ g/ml of Cd ions grew slower than those precultured at 1, 000 $\mu$ g/ml of the metal ions, when they were cultivated in the media containing 1, 000 $\mu$g/ml of the metal ions. Heat shock, however, stimulated the cell growth transiently, when the cells were allowed to grow in the presence of 1, 000 $\mu$g/ml of the metal ions. But the cells given heat shock for more than 100 min received permanent damage to growth. Effects of both stresses on budding rate was also examined. It revealed that the stresses did not change the budding ratio much, which was contradictory to that observed from the same budding yeast, Saccharomyces cerevisiae. Furthermore, the cells treated with 1, 000 $\mu$g/ml of the metal ions not only induced, but also switched off the expression of several new proteins. Some of the cadmium stress-inducible proteins were estimated to be also induced by heat shock stress.