• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1739-1749
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• 2020

Exopolysaccharide produced by the yeast Papiliotrema flavescens, isolated from wine grape berries of Champagne vineyard, was investigated for both chemical and functional characterization. SECMALLS and colorimetric assay analyses showed that the EPS is a high M_W heteropolymer (2.37 × 10⁶ g/mol) majorily consisting of mannose, glucose, xylose and glucuronic acid as monosaccharide constituents, with two substituents (sulphate and phosphate groups), and a minor protein moiety. Structural enchainment of these carbohydrates based on methylation, GC-MS and NMR analyses revealed a linear main backbone built up of α-(1 → 3)-D-mannopyranosyl residues on which are branched side chains consisting of a single β-D-glucopyranosyluronic acid residue and β-(1 → 2)-xylopyranoses (2-5 residues). Suggestion of some xylopyranose side chains containing a mannose residue at the nonreducing terminal end was also proposed. This is first report on EPSs from the grape P. flavescens yeast with such structural characteristics. Furthermore, investigations for valuating the application performance of these EPS in relation with their structural features were carried out in 8% alcohol experiment solutions. Very exceptional viscosifying and foaming properties were reported by comparison with commercial biopolymers such as Arabic, gellan and xanthan gums. The intrinsic properties of the natural biopolymer from this wild grape-associated P. flavescens yeast make it a potential candidate for use in various biotechnology applications.

• Journal of Life Science
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• v.26 no.5
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• pp.545-554
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• 2016

Hyaluronic acid (HA) is an important macromolecule in medical and pharmaceutical fields. HA is a natural and linear polymer composed of repeating disaccharide units of β-1, 3-N-acetyl glucosamine and β-1, 4-glucuronic acid. This work aimed to confirm the structural characteristics and anti-inflammatory activities of HA and its chemically sulfated-HA. HA was produced from a fed-batch fermentation process using Streptococcus dysgalactiae in a 5 l bioreactor. HA was isolated water-soluble form (HA-WS) and water-insoluble form (HA-WI) from culture medium, and was obtained chemically sulfated-derivative (S-HA) that resulted in a 90% yield from HA-WI. The structural features of the sulfated- HA (S-HA) were investigated by FT-IR and ¹H-NMR spectroscopy. The FT-IR and NMR patterns revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. dysgalactiae. The anti-inflammatory activities of HA and S-HA were examined on LPS-induced RAW 264.7 cells. S-HA was significantly inhibited production of pro-inflammatory mediators such as nitric oxide (NO) and PGE₂ and the gene levels of iNOS and COX-2, which are responsible for the production of NO and PGE₂, respectively. Furthermore, S-HA also suppressed the overproduction of pro-inflammatory cytokine TNF-α (<80 pg/ml) and IL-6 (<100 pg/ml) compared to that of HA-WI. The present study clearly demonstrates that HA-S exhibits anti-inflammatory activities in RAW 264.7 macrophage cells.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.218-224
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• 2005

Surface modification of glutaraldehyde fixed bovine pericardium (GFBP) was success­fully carried out with hyaluronic acid (HA) derivatives. At first, HA was chemically modified with adipic dihydrazide (ADH) to introduce hydrazide functional group into the carboxyl group of HA backbone. Then, GFBP was surface modified by grafting HA-ADH to the free aldehyde groups on the tissue and the subsequent HA-ADH hydrogel coating. HA-ADH hydrogels could be prepared through selective crosslinking at low pH between hydrazide groups of HA-ADH and crosslinkers containing succinimmidyl moieties with minimized protein denaturation. When HA­ADH hydrogels were prepared at low pH of 4.8 in the presence of erythropoietin (EPO) as a model protein, EPO release was continued up to $85\%$ of total amount of loaded EPO for 4 days. To the contrary, only $30\%$ of EPO was released from HA-ADH hydrogels prepared at pH=7.4, which might be due to the denaturation of EPO during the crosslinking reaction. Because the carboxyl groups on the glucuronic acid residues are recognition sites for HA degradation by hyaluronidase, the HA-ADH hydrogels degraded more slowly than HA hydrogels prepared by the crosslinking reaction of divinyl sulfone with hydroxyl groups of HA. Following a two-week subcutaneous implantation in osteopontin-null mice, clinically significant levels of calcification were observed for the positive controls without any surface modification. However, the calcification of surface modified GFBP with HA-ADH and HA-ADH hydrogels was drastically reduced by more than $85\%$ of the positive controls. The anti-calcification effect of HA surface modification was also confirmed by microscopic analysis of explanted tissue after staining with Alizarin Red S for calcium, which followed the trend as observed with calcium quantification.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.2
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• pp.285-291
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• 1999

To confirm the food quality of Urechis unicinctus which have been favored as a special raw seafood in southern area in Korea, the chemical composition of Urechis unicinctus and those glycoprotein were studied. Freeze dried Urechis unicinctus was composed of more than 70% of crude protein and 15% of total carbohydrate. The amino acids such as glycine(18.69%), glutamic acid(12.50%) and aspartic acid(9.37%) were noted as major components of total amino acids. The predominant free amino acids were alanine(32.98%), glycine(27.50%), asparagine(19.65%) and taurine(8.29%), and the sum of them were more than 88% to total free amino acids, so they may cause unique taste of Urechis unicinctus with sweetness. As the basis of sugar composition analysis, 56.35% of glucose and 30.49% of N acetylglucosamine were contained respectively, and they might also play an important role in a sweet taste. The leading carbohydrate moiety of glycoprotein from Urechis unicinctus was identified as glucose and N acetylglucosamine similarly to raw muscle, and they occupied more than 50% of total sugar content. Fucose(18.32%) and D glucuronic acid(12.31%) also detected in higher levels com pared to raw muscle. The total amino acid profile of glycoprotein showed a similar trend to raw muscle protein but glycine was detected a significantly lower than that in raw muscle. The glycoprotein from Urechis unicinctus was composed of 4 kinds of subunits, i.e. 25kDa, 20kDa, 18kDa and 12.5kDa of molecular weights through the SDS polyacrylamide gel electrophoresis. On the basis of the IR spectrum of absorptions appeared in 1,240cm-1 and 850cm-1, the glycoprotein had sulfate groups.

• Archives of Pharmacal Research
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• v.17 no.1
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• pp.26-30
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• 1994

Some ammonium oxalate soluble pectic fragments prepared from cultured cell wall of Ephycla distrahya elicited the accumulation of p-coumarocylamino acids (p-CAA) in E. distachya cultures while water soluble and alkali soluble fractions had no activity. Partial purification of the pectic fragments fraction using DEAE-cellulose chromatography afforded two active fractions (PS-I and PS-II) which were composed of mainly uronic acids (98-99 w/w %). They elicited the accumulation of p-CAA in an amount of 52-60 nmol per gram fresh weight of cultures. The acidic sugar compositions of PS-I and PS-II were found to be galacturonic acid and glucuronic acid by TLC analysis. They were supposed to act as endogenous elicitors of p-CAA accumulation. In order to investigate the effect of ethylene on p-CAA accumulation, Ethrel, which is known as ethylene generator, and ACC(1-aminocyclopropane-1-carboxylic acid), a direct precusor of ethylene biosynthesis, were added to the culture. However, they did not glycopeptide elicitor [(Con A-II)], either. Consequently, no relationships between ethylene and p-CAA accumulation were recognized. Several tentative elicitors were teted for their activity. Commercial yeast glucan, $CuCl_2$, laminarin and laminariheptaose had slight activity whereas ${\alpha}$-methylmannopyranoside and commercial yeast mannan had no elicitor activity. ${\alpha}$-methylmannopyranoside which has been known as a tentative inhibitor of glucan elicitor in Glycine max did not affect on the elicitor activity of Con A-II.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.221-227
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• 1990

The aqueous extract of fresh ginseng (Panax ginseng C.A. Meyer) contains a macromolecular fraction that showed mitogenic and co-mitogenic activities in human peripheral blood lymphocytes. Purification of the crude extract by size (ultrafiltration, Sephadex G-200) and charge (DEAE-cellulose, DEAE-Sepharose) yielded a semi.purified fraction (DS-3). This fraction contains at least three subgroups of anionic macromolecules with apparent molecular weight greater than 600 kilodaltons. It is a glycoprotein with a large amount of glucuronic acid. It acts as a mitogen in both T and B cells of human peripheral blood lymphocytes. It could also potentiate the mitogenic action of Concanavalin A in lymphocyte T cells. Such potentiation is not due to increased binding of Concanavalin A to the cell surface. Its mitogenic and co-mitogenic effects do depend on the presence of extracellular Ca2+.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.98-98
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• 1997

This study was done to investigate the effect of vitamin E on hypoxia/reoxygenation-induced hepatic injury in isolated perfused rat liver. Rats were pretreated with vitamin E or vehicle(soybean oil). Isolated livers from fasted 18 hours were subjected to 45min of low flow hypoxia or N$_2$ hypoxia followed by reoxygenation for 30min. The perfusion medium used was KHBB(pH 7.4) and 50${\mu}$㏖/$\ell$ of ethoxycoumarin was added to the perfusate to determine the ability of hepatic drug-metabolizing systems, In low flow hypoxia model, total glutathione and oxidised glutathione levels were significantly increased by hepoxia/reoxygenation with slight increase in LDH levels. These increases were prevented by vitamin E pretreatment. In N$_2$ hypoxia model, LDH, total glutathione and oxidized glutathione levels were increased significantly by hypoxia but restored to normal level by reoxygenation. Vitamin E had little effect on this hypoxic damage. There were no significant changes in the rate of hepatic oxidation of 7-EC to 7-HC in both hepoxic models. But, the subsequent conjugation of 7-HC by sulfate or glucuronic acid were significantly decreased by hypoxia, but restored by reoxygenation in both hypoxia models. As opposed to our expectation, treatment with vitamin E aggrevated the decrease of the rate of conjugation and even inhibited the restoration by reoxygenation. Our findings suggest that hypoxia/reoxygenation diminishes phase II drug metabolizing function and this is, in part, related to decreased energy level.

• Journal of Microbiology and Biotechnology
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• v.20 no.6
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• pp.978-984
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• 2010

This paper examines the probiotic bacterium Lactobacillus rhamnosus GG, and how it reacts to the presence of mucin in its extracellular milieu. Parameters studied included cell clustering, adhesion to mucin, extracellular protein production, and formation of final metabolites. L. rhamnosus GG was found to grow efficiently in the presence of glucose, N-acetylglucosamine, or mucin (partially purified or purified) as sole carbon sources. However, it was unable to grow using other mucin constituents, such as fucose or glucuronic acid. Mucin induced noticeable changes in all the parameters studied when compared with growth using glucose, including in the formation of cell clusters, which were easily disorganized with trypsin. Mucin increased adhesion of the bacterium, and modulated the production of extracellular proteins. SDS-PAGE revealed that mucin was not degraded during L. rhamnosus GG growth, suggesting that this bacterium is able to partially use the glucidic moiety of glycoprotein. This study goes some way towards developing an understanding of the metabolic and physiological changes that L. rhamnosus GG undergoes within the human gastrointestinal tract.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.209-216
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• 2002

Bacillus sp. A56 was studied, because of its high flocculating activity. The flocculating substance produced by this strain was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation, and gel permeation chromatography (GPC). The FT-IR spectrum of the purified bioflocculant, designated as BF-56, showed typical characteristics of polysaccharides. The non-sugar substituents, and sugar components of BF-56 containing glucose, fucose, glucuronic acid, and galactose in an approximate molar ratio of 2.76:1.10:1:0.12, suggested that it was a novel bioflocculant with an estimated molecular mass of over $7{\times}10^3$ kDa. Rheological analysis of BF-56 revealed that it was a pseudoplastic that had higher apparent viscosity rate at dilute concentrations than those of zooglan. The solution of bioflocculant BF-56 exhibited non-Newtonian characteristics and it was compatible to high concentrations of salts such as KCl, NaCl, $CaCl_2,\;or\;FeCl_3.$ The present results suggested strong possibility of bioflocculant BF-56 to be fully applicable to industries such as wastewater treatment.

• Journal of Microbiology and Biotechnology
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• v.30 no.7
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• pp.1092-1096
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• 2020

YjiC, a glycosyltransferase from Bacillus licheniformis, is a well-known versatile enzyme for glycosylation of diverse substrates. Although a number of O-glycosylated products have been produced using YjiC, no report has been updated for nucleophilic N-, S-, and C- glycosylation. Here, we report the additional functional capacity of YjiC for nucleophilic N- and S- glycosylation using a broad substrate spectrum including UDP-α-D-glucose, UDP-N-acetyl glucosamine, UDP-N-acetylgalactosamine, UDP-α-D-glucuronic acid, TDP-α-L-rhamnose, TDP-α-D-viosamine, and GDP-α-L-fucose as donor and various amine and thiol groups containing natural products as acceptor substrates. The results revealed YjiC as a promiscuous enzyme for conjugating diverse sugars at amine and thiol functional groups of small molecules applicable for generating glycofunctionalized chemical diversity libraries. The glycosylated products were analyzed using HPLC and LC/MS and compared with previous reports.