• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.194-201
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• 2014

Clitocybin A is a novel anti-wrinkle cosmetic agent produced by the strain from a Korean native mushroom Clitocybe aurantiaca. In this study, fermentation, extraction, and purification conditions for a large scale production of clitocybin A were optimized, and its cytotoxicity and inhibition activity on the expression of matrix metalloproteinase-1 (MMP-1) were characterized. The mass production of anti-wrinkle agent was achieved according to the 300 L fermentation process with a fed-batch cultivation using the modified yeast-maltose (YM) broth, and a total of 12.5 kg of cell mass was obtained in a 120 L culture broth for 14 days. After extraction and purification, clitocybin A was identified by HPLC. The cytotoxicity of clitocybin A was examined by the MTT assay. When assayed at 100 and 200 ${\mu}g/ml$ concentrations, clitocybin A showed no cytotoxicity, demonstrating safety. The inhibition activity of clitocybin A on the expression of MMP-1 was examined against UV irradiation. Oleanolic acid (control group) showed a relatively low MMP-1 inhibiting activity (ca. 16.7%) at 10 ${\mu}g/ml$ and showed increased cytotoxicity at higher concentrations. In contrast, clitocybin A showed no cytotoxicity at 100 ${\mu}g/ml$, and exhibited a relatively high MMP-1-inhibiting activity (33.1%). These findings indicate that clitocybin A may be a safe and effective anti-wrinkle agent for use in functional cosmetics.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.275-281
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• 1988

To develop cellulase and xylanase-producing strain by protoplast fusion, alkalophilic Bacillus sp. F204 and K17 were treated with NTG(N-methyl-N'-nitro-N-nitrosoguanidine) and isolated anti-biotics resistant strains of S20 (Km$^r$ , Cm$^r$) and G70 (Str$^r$). The frequency of protoplast formation was about 95% when cells of mid-log phase were treated with 200$\mu\textrm{g}$/ml Iysozyme at 37$^{\circ}C$ for 30-45 minutes. Under addition of 0.4-0.5M sodium succinate, 0.5% casamino acid, 1.5% polyvinylpyrrolidone, 25mM MgC1$_2$ and 50mM CaC1$_2$ to the regeneration medium, the regeneration frequency of Bacillus sp. F204 and K17 was 24.9% and 26.2%, respectively. The fusion frequency was 6.6$\times$10$^{-6}$ in the presence of 30% polyethylene glycol 6000 containing 50mM $Ca^{++}$ at 45$^{\circ}C$ for 5 minutes. Cellulase complex and xylanase activities of fusant were compared with parental strains.

• Food Engineering Progress
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• v.13 no.3
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• pp.183-189
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• 2009

Lactobacillus sakei B2-16 producing high level of $\gamma$-amino butyric acid (GABA) was previously isolated from Korean traditional fermented food, Kimchi. L. sakei B2-16 converted 99.3% of mono sodium glutamate (MSG) to GABA in Lactobacilli MRS broth supplemented with 1% MSG. In order to enhance the production of GABA by L. sakei B2-16, growth parameters such as media components and concentrations of major components were evaluated. The maximum GABA concentration was obtained by a modified rice germ extract broth containing 4%(w/v) sucrose and 1%(w/v) yeast extract. L. sakei B2-16 converted 100% of MSG to GABA in modified rice germ extract broth supplemented with 7% MSG.

• Applied Biological Chemistry
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• v.17 no.2
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• pp.93-116
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• 1974

A potent intracellular-lipid-producing yeast, Rhodotorula glutinis var. glutinis SW-17, was screened out from a variety of arable soils, compost heaps, and fodders, and two strains of excellent extracellular-lipid-producing yeasts, Rhodotorula glutinis var. glutinis SW-5 and Rhodotorula graminis SW-54, were screened out from the surface of many species of leaves. And then the intra- and extra-cellular lipid productions by those Rhodotorula yeasts were studied. The results were as follows: 1. During the shaking culture of 8 days at $24^{\circ}C$, both the intra- and extra-cellular lipid accumulation started almost at the stationary phase of growth, when the nitrogen source in the medium was a little more than half used up. The intracellular lipid production by Rhodotorula glutinis var. glutinis SW-17 reached 58.42% (w/w) of dried yeast, and the extracellular lipid production by Rhodotorula graminis SW-54 amounted to 2.62g per liter of the medium. 2. After the carbon and nitrogen sources in the medium were almost consumed, if the yeasts were shake-cultured further in a state of starvation, the yeast cells re-utilized the already produced intra- and extra-cellular lipids and the lipids completely disappeared in the medium in about 90 days. 3. The relative concentration of carbon and nitrogen sources in the media greatly influenced both the intra- and extra-cellular lipid production. When the nitrogen source in the medium was almost used up for the growth of yeast, and excess carbon sources were still available, the lipid production vigorously proceeded. As long as the nitrogen source concentration in the medium was high, the lipid production was greatly suppressed. 4. The optimum pH for both the intra- and extra-cellular lipid production by those yeasts was pH 5.0-6.0. 5. The fatty acid components of the intracellular lipid of Rhodotorula glutinis var. glutinis SW-17 were myristic, palmitic, palmitoleic, stearic, oleic, linoleic, and linolenic acids. The largest components of the fatty acids were palmitic acid equivalent to 30-45% of the whole fatty acids and oleic acid equivalent to 35-50%. 6. The fatty acid components of the extracellular lipid of Rhodotorula glutinis var. glutinis SW-5 and Rhodotorula graminis SW-54 were myristic, palmitic, stearic, oleic, linoleic, linolenic, 3-D-hydroxypalmitic, and 3-D-hydroxystearic acids. The largest components of the fatty acids were 3-D-hydroxypalmitic acid equivalent to 22-25% of the acids and 3-D-hydroxystearic acid equivalent to 13-17%. 7. The polyol component of the intracellular lipids was only glycerol, whereas the polyols of extracellular lipids were glycerol, mannitol, xylitol and arabitol.

• Microbiology and Biotechnology Letters
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• v.3 no.3
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• pp.123-134
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• 1975

Out of 96 yeast strains isolated from various natural habitats, five strains were screened based on their ability to ferment agricultural biproducts such as rice-, barley-and wheat-bran, and sawdust. These were identified as Hansenula anomala var anomala, Candide utilis, C. pelliculosa, Debaryomyces hansenii, and Irpex lacteus. Using these yeasts the above mentioned agricultural biproducts were fermented in various combinations. The fermented product was fed to 180 male Starcroses for eight weeks and obtained a body weight increase of 15.1g a day, while the unfermented control feed increased 10.5g a day.

• Microbiology and Biotechnology Letters
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• v.8 no.3
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• pp.181-191
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• 1980

For the purpose of studies on the citric acid production, some experiments were carried out with isolated strains. The results obtained were as follows. 1) The optimal culture media of the strain M-80 in surface culture contained 140g of sucrose, 3.0g of (N $H_4$)$_2$S $O_4$, 1.5g of K $H_2$P $O_4$, 0.2g of MgS $O_4$.7$H_2O$, 3.0mg of F $e^{++}$, 1.0mg of Z $n^{++}$, 0.5N HCI to a pH of 5.0 and distilled water to 1.0 liter; and that of the strain M-315 in surface culture contained 140g of sucrose, 2.0g of N $H_4$N $O_3$, 1.0g of K $H_2$P $O_4$, 0.25g of MgS $O_4$. 7$H_2O$, 2.0mg of F $e^{++}$, 2.0mg of Z $n^{++}$, 0.05mg of C $u^{++}$, 0.5N HCI to a pH of 4.5 and distilled water to 1.0 liter. While that of the strain M-315 in submerged culture contained 140g of sucrose, 2.5g of N $H_4$N $O_3$, 1.5g of K $H_2$P $O_4$, 0.3g of MgS $O_4$. 7$H_2O$, 3.0mg of F $e^{++}$, 0.1mg of C $u^{++}$, 0.5N HCI to a pH of 4.5 and distilled water to 1.0 liter. The optimal temperature and size of inoculum were mostly 28-3$0^{\circ}C$, 10$^{7}$ -10$^{8}$ spores/50ml, respectively. 2) Through the course of citric acid production, the growth of strains had nearly been completed, pH value was rapidly decreased below 2.0 and the content of sugar was also reduced, while the accumulation of citric acid in media was remarkably begun in about 3-4 days. The yields of citric acid generally reached the maximum level in 8-10 days in surface or submerged fermentation process. 3) Methanol was effective citric acid production when they were added to fermentation media. In the case of surface culture, by addition of 2％ (strain M-80), 3％ (strain M-315), the yields of citric acid was increased 6.5％, 20.6%, respectively and 5.0% yield was increased by addition of 3% methanol in submerged culture media of the strain M-315. 4) Chromatography analysis of culture broth after fermentation under optimal culture conditions detected that the majority of acid in media was citric acid. 72.1mg/ml, 98.1mg/ml, of citric acid were determined in surface culture media by strains of M-80, M-315, and 59.8 mg/ml of citric acid was contained in the submerged culture media by the strain M-315. strain M-315.