• Korean Journal of Microbiology
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• v.24 no.1
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• pp.57-61
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• 1986

In order to remove of cadmium from waste water an identification of a cadmium-ion tolerant yeast B-7 isolated from the sludge of zinc mining district was studied. By the taxonomecal characteristics of strain B-7 it was identified as Hansenula anomala B-7 or similar strain. The cadmium-ion tolerance of the strain B-7 was determined as $2,700{\mu}g/ml$ of cadmium-ion by density gradient agar plate method. The strain B-7 grew well in an aqueous medium containing $1,000{\mu}g/ml$ of cadmium-ion.

• The Korean Journal of Mycology
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• v.19 no.4
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• pp.282-284
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• 1991

The yeast, Hansenula anomala B-7, isolated from the $Cd^{2+}$ rich soils and determined to be tolerant in the high concentration of $Cd^{2+}$ were employed in this work. Its intact cells grown in high concentration of $Cd^{2+}$ were observed to be Iysed at the early stage when transferred to a cadmium deficient broth. Its intact cells found to be not Iysed and grow well under the high concentration of $Cd^{2+}$. The Iysis of the intact cells grown at the high concentration of $Cd^{2+}$ ion was not found when the metal ions were replaced with $Cd^{2+}$ ion in the same concentration. This result indicated that Iysis of yeast cells, at least in this isolate, would be related to cell osmosis with the mineral ions added.

• Korean Journal of Microbiology
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• v.29 no.6
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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.

• Korean Journal of Microbiology
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• v.25 no.2
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• pp.110-116
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• 1987

As a pary of investgation on effective accumulation of cadmium in yeast cells, effect of surfactant was studied on intracellular accumulation of cadmium in extremely cadmium-tolerant yeast, Hansenula anomala B-7. Cadmium accumulation was enhanced up to approximately 40% by the addition of non-ionic surfactant, Triton X-100 and its optimal concentration was found to be 0.1-0.2%. Furthermore, optimum conditions for intracellular accumulation of cadmium were at $40^{\circ}C$ and initial pH 6.0 for 48 hours under shaking culture.

• Korean Journal of Microbiology
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• v.28 no.3
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• pp.258-263
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• 1990

A cadmium-binding protein was purified the cell-free extract of extreme cadmium tolerant Hansenula anomala B-7. The molecular weight was determined to be approximately 33, 000 and was composed two kinds of subunits having a molecular weight of 18, 000 and 14, 000, respectively. The extinction coefficient of the cadmium-binding protein was calculated to be 19.58. The amount of cadmium in the cadmium-binding protein was $9.26{\mu}{\textrm{g}}$ per $100{\mu}{\textrm{g}}$ of protein. A total of 14 amino acids were detected in the cadmium-binding protein, including aspartic acid, glycine and alanine that were present in a high quantity, but proline, valine and methionine were not found. The purified cadmium-binding protein contained a high quantity of cysteine and cadmium, and therefore this protein showed clearly the characteristics of metallothionein.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.239-243
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• 1990

In this study the authors investigated the distribution of cadmium accumulated in cadmium-iun tolerant Hansenula anomala B-7 cells and also the effect of cadmium on protein synthesis. 84.9% of the cadmium accumulated was distributed in the soluble fraction (cytosol, etc.). The intracellular protein content was decreased by cadmium (1,000 $\mu g$/ml), but the content of soluble protein preeipitated by ammonium sulfate (30-75% saturation) was increased compared with the content of it obtained from the cells grown without cadmium. Furthermore, in the cells grown with 1,000 $\mu g$/ml of cadmium t h higher molecular weight soluble protein was increased compared with the cells grown without caa, mium, but the lower molecular weight soluble protein was decreased. These results suggested that the protein synthesis was inhibited by cadmium, but synthesis of higher molecular weight soluble protein was remarkably stimulated by cadmium.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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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.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.225-230
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• 1998

This study was carried out to investigate the effect of cadmium ion on activities of cadmium-adapted malate dehydrogenase (adapted-MDH), which is defined to be an enzyme obtained from an extreme cadmium-tolerant yeast Hansenula anomaul B-7 grown in medium containing 1 mM cadmium ion. Cadmium-nonadapted malate dehydrogenase (nonadapted-MDH), which is defined to be enzyme expressed in the cells grown in $Cd^{2+}$ -free medium was also characterized by the same manner. Activities of the adapted-MDH and the nonadapted-MDH were strongly induced to 450% and to 150% in comparision with the control examined with 1 mM $Cd^{2+}$, respectively. The adapted-MDH activity was stimulated to 147%, 150%, and 135% compared with the control analyzed with 1 mM $Zn^{2+}$, 1 mM $Mn^{2+}$, and 1 mM $Ca^{2+}$, respectively and to 925%, and 250% compared with the control analyzed in the presence of 2 mM $Cd^{2+}$, and 2.5 mM $Zn^{2+}$, respectively. Km values of the adapted-MDH and the nonadapted-MDH were calculated to be the same 6.9 mM for L-malate, respectively. The Km value of the nonadapted-MDH was not changed by $Cd^{2+}$ while Vmax of the nonadapted-MDH was increased by $Cd^{2+}$. In contrast, both the Km and the Vmax values of the adapted-MDH were changed by $Cd^{2+}$.

• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.29-34
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• 1989

An intracellular accumulation of cadmium by the intact cell of an extremely cadmium tolerant yeast, Hansenula anomala B-7, was investigated in the presence of Triton X-100. The uptake of cadmium by the intact cell was efficiently enhanced up to approximately 40% or more by 0.1% of Triton X-100 and Aerosol OT, respectively. The Michaelis constant, Km, done by Lineweaver-Burk plot of accumulation velocity of cadmium vs. cadmium concentration was calculated to be 0.247mM. The optimal conditions of pH and the temperature for the effective cadmium uptake were from neutrality to alkali and 4$0^{\circ}C$, respectively. The accumulation of cadmium was increased approximately 3 times under the shaking incubation, with no correlation to shaking rate. By zinc the cadmium accumulation was decreased.

• Applied Biological Chemistry
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• v.13 no.2
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• pp.171-180
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• 1970

The yeasts in the soy sauce mash were isolated and identified, and they were classified by coloring with the treatment of TTC(2, 3, 5, triphenyltetrazolium chloride) agar and counted in process of time. The results obtained were as follows: a) The number of ordinary and osmophilic yeasts in 1 ml. of the soy sauce mash showed a tendency to be increased from the mashing to the mature stages and to decrease in the aging stages: $127{\times}10^3$ immediately after mashing, $83{\times}10^3$ 1 month after, $356{\times}10^3$ 3 months after, $1250{\times}10^3$ 6 months after and $65{\times}10^3$ 2 years after mashing in the case of ordinary yeasts, and 0 after mashing, $40{\times}10^3$ 1 month after, $81{\times}10^3$ 3 months after, $358{\times}10^3$ 6 month after and $23{\times}10^3$ 2 years after mashing in the case of osmophilic yeasts. b) 50 strains of yeasts were isolated from the soy sauce mash optionally in process of fermentation period, and they were identified as 7 genera and 18 species: 10 strains of Saccharomyces rouxii, 1 strain of Saccharomyces marxianus, 3 strains of Saccharomyces rosei, 1 strain of Saccharomyces fermentati, 6 strains of Saccharomyces mellis, 1 strain of Saccharomyces acidifaciens, 1 strain of Saccharomyces pastori, 3 strains of Pichia polymorpha, 2 strains of Hansenula anomala, 1 strain of Hansenula saturnus, 2 strains of Hansenula suaveolens, 5 strains of Nadsonia fulvescens, 8 strains of Debaryomyces hasenii, 1 strain of Debaryomyces nicotianae, 1 strain of Debaryomyces kloeckeri, 2 strains of Torulopsis sake, 1 strain of Torulopsis holmii and 1 strain of Candida pelliculasa. c) Distribution of yeasts according to the fermentation period was as follows: i) Saccharomyces rouxii, Saccharomyces marxianus, Saccharoymces rosei, Pichia polymorpha, Debaryomyces hansenii, Torulopsis sake, Candida pelliculosa, Debaryomyces nicotianae, Nadsonia fulvescens, Hansenula suaveolens and Hansenula saturnus were found in the early stages of fermentation. ii) Saccharomyces rouxii, Saccharomyces rosei, Saccharomyces fermentati, Saccharomyces mellis, Saccharomyces pastori, Hansenula anomala, Saccharomyces acidifaciens and Debaryomyces hansenii appeared in the mature stages. iii) Saccharomyces rouxii, Saccharomyces mellis, Nadsonia fulvescenes, Dedaryomyces hansenii, Debaryomyces kloeckeri, Torulopsis sake and Torulopsis holmii were distributed in the aging stages. d) TTC white yeasts were found in abundance in the early stages of fermentation and TTC red yeasts appeared more than 50 per cent in the mature and aging stages. e) The yeasts belonging to Saccharomyces mellis and Saccharomyces pastori were classified as TTC red yeasts, Saccharomyces acidifaciens were reel pink, Hansenula saturnus Debaryomyces kloeckeri, and Torulopsis holmii were pink, Saccharomyces marxianus and Nadsonia fulvescens were white and the others were the same as the description in the previous report. Saccharomyces rouxii ware classified for the most part as TTC red yeasts, and while some of them were red pink. f) Species of yeasts in the soy sauce mash were similar to those in the soy sauce koji, but the latter were not osmophilic and in the former case, the osmophilic yeasts were increased in process of fermentation period.