• Korean Journal of Microbiology
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• v.37 no.1
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• pp.42-48
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• 2001

Allele rescue and sequence analysis of soo1-1 allele in Saccharomyces cerevisiae mutant LP0353 revealed that soo1-1 is identical to the previously reported ret1-1 allele, which has a base substitution of A for $G^{681}$ leading to an amino acid substitution of aspartic acid for $glycine^{227}$ in Soolp. However, it was revealed that the addition of osmotic stabilizer, such as 1.2M sorbitol can rescue the temperature sensitive phenotype of the ret1-1 mutant and that the soo1-1/ret1-1 mutation may confer defects in post-translational modification of proteins involved in the yeast cell wall biogenesis. Evidence for a putative role of 5th WD40 domain of the Soo1p/$\alpha$-COP in the construction and maintenance of cell walls was also presented by complementation test with deletion constructs of the SOOl.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.620-628
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• 2012

We examined the effects of biological activity Phellinus linteus mycelium culture with cassiae semen extract. Firstly, the optimal temperature, initial pH and culture period for mycelial growth in a liquid culture of P. linteus were determined, and they were $30^{\circ}C$, pH 5.0 and 8 days respectively. The five herbal materials were examined against several health functional efficacies, and, as a result, Cassiae semen was chosen, with its superior inhibitory effects in ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities(95.3%, 80.9%, 96.1 and 24.2%, respectively). P. linteus fruit body was investigated on ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities, and they were 54.7%, 81.9%, 30.0% and 20.1%, respectively. Accordingly, C. semen was used in the following experiment, to give an additive functional effect on the P. linteus. As the amount of C. semen in the cultural media increased, mycelial weight and ${\beta}$-glucan contents also increased, but final pH was not influenced. In addition, the ${\beta}$-glucuronidase inhibitory activity, electron donating activity, and ${\alpha}$-glucosidase inhibitory activity increased. P. linteus mycelium culture showed higher activities in the other three tests above, except for electron donating activity, when C. semen was added to the medium before cultivation.

• Journal of Microbiology
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• v.37 no.2
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• pp.80-85
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• 1999

The purification system of glucoamylase (glucan 1,4-${\alpha}$-glucosidase, EC 3. 2. 1. 3), some characteristics of the purified enzyme and hydrolysis rate of various raw starch were investigated through several experiments. The enzyme was produced on a solid, uncooked wheat bran medium of Aspergillus oryzae NR 3-6 isolated from traditional Korean Nuruk. The enzyme was homogeneously purified 6.8-fold with an overall yield of 28.3% by the criteria of disc- and SDS-polyacrylamide gel electrophoresis. The molecular weight was estimated to be 48 kDa by SDS-PAGE. The optimum temperature and pH were 55$^{\circ}C$ and 4.0, respectively. The enzyme was stable at a pH range of 3.0∼10.0 and below 45$^{\circ}C$. Enzyme activity was inhibited about 27% by 1mM Hg2+. The hydrolysis rate of raw wheat starch was shown to be 17.5-fold faster than the hydrolysis rate of soluble starch. The purified enzyme was identified as glucoamylase because the product of soluble starch by the purified enzyme was mainly glucose by thin layer chromatography.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.57-67
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• 2011

Agaricus brasiliensis, one of edible mushroom belonging to Basidiomycota, has been used for curing gastric ulcer and stomach cancer of human beings and also known to have good inhibitory effects on sarcoma 180 and Ehrlich carcinoma of mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were prepared from fruiting body of the mushroom. ${\beta}$-glucan and total protein contents were identify from fractions of edible mushrooms extract. The ${\beta}$-glucan and protein contents of all fractions of the mushrooms ranged from 21.54~32.31% and 0.16~9.34%, respectively. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, NIH3T3 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 18.8~50.6% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cell by 1.2 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. MeOH and Na improved the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.6 fold compared with control at the concentration of 50~500 ${\mu}g/ml$. Fr. Na generated 15.9 ${\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7 at the concentration of 200 ${\mu}g/ml$, while lipopolysaccharide, a positive control, produced 3.7 ${\mu}M$. The Fr. NaCl, Fr. HW and Fr. MeOH increased the secretion of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by 2.2 times compared with the control group. Fr. Na increased the numbers of peritoneal exudate cells by 4 folds at the concentration of 50mg/kg compared with control. Circulating leukocytes increased by 2.7 folds when Fr. HW from A. brasiliensis was inoculated at the concentration of 50 mg/kg body weight. The hematological and blood chemical analysis of the 3 fractions did not show any difference in blood compositions and enzyme activities compared with the control group (p<0.05). Therefore, the experimental results suggested that crude polysaccharides extracted from A. brasiliensis contain antitumor and immuno-potentiating activities against Sarcoma 180 in ICR mice.

• IMMUNE NETWORK
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• v.4 no.2
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• pp.94-99
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• 2004

Background: We previously reported that ginsan, a polysaccharide extracted from Panax ginseng had an immunostimulatory activity such as mitogenic activity, activation of macrophages and killer cells, and production of a variety of cytokines which resulted in antitumor and antiseptic effects. We further purified $\alpha$-(1$\longrightarrow$6)-glucan and $\beta$-(2$\longrightarrow$6)-fructan from the ginsan with size exclusion and ion-exchange column chromatography successively. In this study, we performed the structure-based activity of ginsan by comparison with known polysacchrides such as $\beta$-glucan, curdlan, laminarin, levan, dextran, lentinan and OK-432. Methods: To investigate the immunostimulatory activity of several polysaccharide compounds, we investigated the stimulation of lymphocytes proliferation, the generation of activated killer cells and the secretion of nitrites from activated macrophages. Results: Of polysaccharides tested, curdlan and ginsan stimulated lymphocyte proliferation, suggesting that the molecular weight and composition of polysaccharide are dependent on the mitogenic activity. The production of nitric oxide was significantly increased in curdlan, levan, ginsan and its fraction, indicating that fructan has also capacity to activate macrophages and may devote to kill pathogens. In addition, the activation of macrophages was seemed to be independent of molecular weight of polysaccharide. The generation of AK cells was exhibited in order of curdlan, OK-432> F1, ginsan, F3> levan> etc. The AK activity may be dependent on molecular weight and composition of polysaccharides. Conclusion: Unfortunately, purified polysaccharide from ginsan were less active on immunostimulatory activity than mixed compounds of polysaccharides. From the viewpoint of structure and activity relationships, we found several characteristic features.

• Journal of the Korean Wood Science and Technology
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• v.25 no.3
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• pp.1-7
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• 1997

한국산 동백나무겨우살이(Pseudixus japonicus Hayata)에 존재하는 수용성 리그닌-탄수화물 복합체를 구성하는 다당류의 구조를 밝히고 리그닌 성분과 다당류의 결합양식을 구명하고자, 냉 온수 추출한 수용성 리그닌-탄수화물 복합체(M-LCC-WE)를 DEAE Sephadex A-50로 중성분획(M-LCC-N)과 산성분획(M-LCC-A), 나머지분획(M-LCC-R)으로 세분화한 후 M-LCC-N과 M-LCC-A에 대하여 메틸화, 아세틸화, 그리고 DDQ 산화반응을 실시하였다. M-LCC-N을 구성하는 다당류는 ($1{\rightarrow}4$) 글리코시드결합의 arabinan과 ($1{\rightarrow}4$)나 ($1{\rightarrow}6$) 글리코시드결합의 galactan과 glucan으로 M-LCC-A의 다당류는 ($1{\rightarrow}4$) 글리코시드결합의 arabinan과 ($1{\rightarrow}6$) 글리코시드결합의 galactan이 다당류 주성분으로 밝혀졌으며 galacturonic acid가 결합되어 있기 때문에 산성적 성질을 나타내고 있었다. 또한 M-LCC-A에서는 galacturonic acid 의 carboxyl 그룹이 리그닌의 ${\alpha}$-와 ${\gamma}$-위치에서 ester결합이 존재함이 확인되었다.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.125-132
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• 2006

The below is the results of physicochemical analysis of 40 breeding lines of colored barley (CB) whose systems are different Water content of CB No. 24 showed the lowest value of 7.4% while CB No. 9 showed the highest value of 10.8%. Crude protein of CB $9.7{\sim}12.9%$ range was found. Crude fat content of CB No. 6 showed the highest value of 4.35% while CB No. 34 showed the lowest of 1.35%. Crude ash content of CB No. 31 showed the lowest value of 1.20%. Ca content of CB No. 10 showed the highest value of 717.50 mg% while general barley showed the lowest value of 442.82 mg%. Mg content of CB No. 10 showed the highest value of 1320.00 mg%. Cu content of CB No. 20 showed the lowest value of 2.20 mg% while CB No. 33 showed the highest value of 6.25 mg%. K content of CB No. 20 showed the lowest value of 723.24 mg% while CB No. 1 showed the highest value of 1002.50 mg%. Mn content of CB No. 28 showed the lowest value of 31.72 mg% while general barley showed the highest value of 94.56 mg%. ${\beta}-Glucan$ content of CB No. 25 showed the lowest value of 5.20 mg% while CB No. 28 showed the highest value of 4.46 mg%.

• Journal of the korean academy of Pediatric Dentistry
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• v.23 no.3
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• pp.764-773
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• 1996

The mutan containing $\alpha$-1,3 bond is an insoluble portion of glucan which is the main component of dental plaque. The secretion of mutanase was assessed with mutan-digesting Streptomyces isolated from soil, and the factors affecting its activity was studied, obtaining the following result. Mutan-digesting Streptomyces was identified as Streptomyces exfoliatus by its characteristics. The effect of dextranase was identified on the media containing blue dextran. A clear zone was produced by Streptomyces exfoliatus on the media containing blue mutan, so showing the secretion of mutanase. A clear zone was significantly produced on the media overlayed with agar containing blue mutan. A clear zone was produced at 2 days after the inoculation of Streptomyces exfoliatus on the media containing below a concentration of 0.025% glucose, at 3 days on the media containing 0.05 % glucose, and at 4 days on the media containing 0.1 % glucose. Mutan-digestion wasn't appeared early by adding other carbohydrates. The higher concentration of peptone, the later appearance of clear zone was on the media containing below a concentration of 0.1 % peptone. These results indicated that the secretion of mutanase was identified from mutan-digesting Streptomyces on the media containing blue mutan, and a clear zone was appeared lately on the media containing higher amount of glucose.

• The Korean Journal of Food And Nutrition
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• v.3 no.2
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• pp.149-160
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• 1990

The soybean $\beta$-amylase ($\alpha$-1, 4-glucan maltohydrolase, EC 3.2.1.2) is composed of seven isozymes(I', I, II, III, IV, V and VI), and isozyme II and IV are the main components among these. The Purification of $\beta$-amylase isozymes from soybean whey were performed by ammonium sulfate fractionation, CM-Sephadex C-50 column chromatography, DEAE-Sephadex chromatography and Gel filtration. The resulted purity of $\beta$-amylase was throughly confirmed by electrophoresis, and then determined its isoelectric point and molecular weight. The results obtained were as follows, 1. Five active fractions of soybean p-amylase were derived on CM-Sephadex C-50 column chromatography. 2. Seven active bands of p-amylase isozymes were detected by isoelectric focusing gel electrophoresis, and their isoelectric points(I' to VI) were 5.07, 5.15, 5.25, 5.40, 5.55, 5.70 and 5.93, respectively. 3. Isozyme II and IV were main components of soybean $\beta$-amylase. 4. The molecular weights of both isozyme II and IV were determined to be 56,000 daltons by the result of SDS polyacrylamide gel electrophoresis. 5. Km values of main isozyme II & IV for amylopectin were determined to be 2.25 mg/ml, which suggest the same function of each isozyme.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.644-648
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• 2005

Dextranase (${\alpha}$-1,6-D-glucan-6-glucanogydrolase:E.C. 3.2.1.11) catalyzes the hydrolysis of ${\alpha}$-(1.6) linkages of dextran. A lsd1 gene encoding an extracellular dextranase was isolated from the genomic DNA of L. starkeyi. The lsd11 gene is a synthetic dextranase (lsd1) after codon optimization for gene expression with Pichia pastoris system. A open reading frame of lsd11 gene was 1827 bp and it was inserted into the pPIC3.5K expression vector. The plasmid linearized by Sac I was integrated into the 5'AOX region of the chromosomal DNA of P. pastoris. The lsd11 gene fragment encoding a mature protein of 608 amino acids with a predicted molecular weight of 70 kDa, was expressed in the methylotrophic yeast P. pastoris by controling the alcohol oxidase-1 (AOX1) promoter. The recombinant lds11 was optimized by using the shake-flask expression and upscaled using fermentation technology. More than 9.8 mg/L of active dextranase was obtained after induction by methanol. The optimum pH of LSD11 was found to be 5.5 and the optimum temperature $28^{\circ}C$.