• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.179-186
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• 1998

Dextransucrase hyper-producing Leuconostoc mesenteroides B-512FMCM and dextransucrase constitutive mutants B-742CB and B-1355C catalyzed the transfer of glucose from sucrose to other carbohydrates which were present or were added to the reaction digests. When the acceptor was a maltose, gentiobiose, lactose or raffinose, there was produced a series of oligosaccharide acceptor products or single product based on the kinds of enzymes and reaction conditions. To obtain the quantitative information about the yield and the distribution of acceptor products and dextran two experimental parameters were studied: a) the ratio of acceptor to sucrose and b) the amount of enzyme at constant carbohydrate concentration (100 mM). As the amount of enzyme increased, the synthesis of acceptor products (of maltose or gentiobiose) increased, and the formation of dextran decreased. As the ratio of acceptor to sucrose increased, the amount of dextran and the number of acceptor-products decreased and the amount of acceptor-products increased. When maltose or gentiobiose was an acceptor, the glucose from sucrose was transferred to the C-6 hydroxyl group of the nonreducing-end glucose residue of accepters to give a homologous series of isomaltosyl dextrins. In case of lactose or raffinose, there was produced only one acceptor product from B-512FMCM dextransucrase reaction. In the lactose acceptor reaction, the glucose from sucrose was transferred to the C-2 hydroxyl of the reducing end glucose residue of lactose. To get a series of oligosaccharides from lactose or raffinose acceptor reaction we used B-742CB dextransucrase or B-1355C alternansucrase with 500 mM sucrose in reaction digest.

• Korean Chemical Engineering Research
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• v.47 no.4
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• pp.506-511
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• 2009

In order to improve the bacterial cellulose(BC) production yield, centrifugal and inclined centrifugal impellers were developed. A 6 flat-blade turbine impeller was used as a control system. The flow pattern in the fermenter and volumetric oxygen transfer coefficient($k_La$) of these fermentation systems were studied. Fermentations were carried out for the production of BC by G. hansenii PJK in a 2-L jar fermenter equipped with new impellers. Liquid medium was circulated from the bottom, through the cylinder of the impeller and to the wall. The volumetric oxygen transfer coefficients, $k_La$, of inclined centrifugal and centrifugal impeller systems at 100 rpm were 23 and 15% of the conventional turbine impeller system, respectively. However, the conversion of microbial cells to cellulose non-producing mutant decreased and this results in the increase in BC production at low rotating speed of impellers.

• Journal of Dairy Science and Biotechnology
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• v.23 no.2
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• pp.155-160
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• 2005

Many countries divide infant formula into special nutrient food and they are in control of thorn very carefully. CODEX prescribes that the components which can be added to infant formula must be exist in the breast milk and the additives must be scientifically proved to be safe. Therefore infant formula manufacturers make efforts to develop infant formula to be similar to human milk based on research of human and bovine milk components. Domestic infant formulas are rich with functional components in comparison with foreign formulas ; such as immunity, anti-allergy, growth factor, developments of brain and digestion organs etc. At this point, progress of bioscience technology and introduction of various new technologies lead micro components in human and bovine milk to be separated and purified. Therefore many functional components are commercialized and infant formula companies use these. Various functional components are added to infant formula ; DHA and arachidonic acid as brain growth components, nucleotide and lactoferrin as immunity increment and oligosaccharides fur digestion. And new infant formula is being developed by using bone growth factor and dehydration components f3r atopy and allergy decrement.

• Applied Biological Chemistry
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• v.39 no.2
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• pp.99-103
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• 1996

A bacterial strain producing a novel endo-inulinase, hydrolysing inulin into oligosaccharides was isolated from soil and identified as Arthrobacter sp. S37 The enzyme production was induced by inulin and jerusalem artichoke extract. The maximum enzyme production was obtained with medium containing 1.5% jerusalem artichoke extract, 1.0% yeast extract, $0.5%\;NaNO_3,\;0.05%\;MgSO_4{\cdot}7H_2O,\;0.05%\;KCl,\;0.0016%\;FeCl_3{\cdot}6H_2O\;and\;0.05%\;KH_2PO_4$. The optimum temperature and pH for the enzyme production were $30^{\circ}C$ and 8.0, respectively. Under the optimum condition, the enzyme activity in the culture broth reached at maximum, 10.8 units/ml after cultivation for 24 hours.

• Journal of the Korean Wood Science and Technology
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• v.15 no.3
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• pp.34-43
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• 1987

This study was performed to find out the types of linkage of carbohydrates in wood cell walls. To study the structure of linkage of carbohydrates in wood cell walls, we have attempted to find out the method holocellulose preparation and optimum condition of enzyme hydrolysis in holocellulose, and fractionate oligosaccharide with products that hydrolized partly by acetolysis and deacetylation in holocellulose. We have achieved four results. These results as follow; 1. At first. we reacted in wood meal $NaClO_2$ 1g per lignin lg for one hour and then the same of quantity $NaClO_2$ for four hours. Through these experiments, we have developed new holocellulose preparation method which had low loss of carbohydrates and high effect of the delignification. 2. The optimum condition of enzyme hydrolysis of holocellulose which had lignin was 0.005M sodium acetate buffer (pH 5.0). We have achieved 7.2% reducing sugar through the procedure that reactioned 0.01g holocellulose putting enzyme 0.03g for 72 hours. It may be supposed that 5.5% of lignin contained in holocellulose prevented enzyme contaction from holocellulose and so this lignin has resulted in the low efficiency of enzyme hydrolysis. 3. We did not fractionated from oligosaccharides which were preparated by the method of acetolysis and deacetylation in holocellulose. The reason is that holocellulose having a lot of lignin prevented prefectly partial hydrolysis from the method of acetolysis and deacetylation. 4. We attempted analysis of six standard substances through HPLC apparatus having sugar pak 1 column which we have changed flow rate and the column temperature variably. These six standard substances were D-glucose, D-mannose, D-xylose, D-galactose and L-rhamnose, L-arabinose, But sugar pak 1 column was not fitted analysis of four substances because D-galactose, D-mannose, D-xylose, L-rhamnose were agreement with elution time. And so, we could not analize four standard substances with sugar pak 1 column.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.352-373
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• 2003

Rhamnose-rich (RROP-s) and fucose-rich (FROP-s) oligo-and polysaccharides were prepared and extensively characterised by physical and chemical procedures [1,2] and compared to L-fucose. Their biological properties were then studied on human skin fibroblast cell cultures, human skin explant cultures and on hairless rat skin, using a variety of cell-biological, biochemical and computerised morphometrical procedures. Among the most important properties we could establish, the following are of particular interest for the tretment and prevention of age-dependent modifications of human skin (loss of skin-tissue, cells and matrix, wrinkle formation and others) : stimulation of cell proliferation (by $^3$[H]-thymidine incorporation and the MTT test), scavenging of reactive oxygen species (ROS) using several different procedures, and protease (MMP-2 and MMP-9) down-regulation. A topical preparation, using RROP-s and FROP-s, and/or L-fucose, was shown to increase cell proliferation, dermal matrix synthesis, efficient scavenging of ROS-s and to increase also the thickness of dermal tissue when applied for 4 weeks on hairless rat skin, accompanied by the densification of collagen bundles as well as by an increase of elastin synthesis. Using fluorescent labeled FROPs, it could be shown that these oligosaccharides react with cell-membrane receptors and especially with the elastin-laminin-receptor and the fucose-mannose receptor, but they penetrate also in the cell nucleus, suggesting the possibility of a direct action on the regulation of gene expression. When applied to the human skin of a team of voluntary women encompassing all age-groups, the efficiency of FROP-containing preparation could be confirmed using indentometry and computerised evaluation of skin micro-relief, as well as evaluation of periorbital wrinkles. It appears therefore that these preparations correspond to all the requirements of active anti-aging principles.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1546-1554
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• 2010

A bacterial strain capable of producing extracellular ${\alpha}$-galactosidase was isolated from a sample of sugarcane industrial waste. Microbiological, physiological, and biochemical studies revealed that the isolate belonged to Bacillus sp. Furthermore, based on a 16S rDNA sequence analysis, the new isolate was identified as Bacillus megaterium VHM1. The production of ${\alpha}$-galactosidase was optimized based on various physical culture conditions. Guar gum and yeast extract acted as the best carbon and nitrogen sources, respectively. The optimum pH was 7.5 and the enzyme remained stable over a pH range of 5-9. The enzyme was optimally active at $55^{\circ}C$ and thermostable with a half-life of 120 min, yet lost 90% of its residual activity within 120 min at $60^{\circ}C$. One mM concentrations of $Ag^2$, $Cu^2$, and $Hg^{2+}$ strongly inhibited the ${\alpha}$-galactosidase, whereas the metal ions $Fe^2$, $Mn^{2+}$, and $Mg^{2+}$ had no effect on the ${\alpha}$-galactosidase activity, and $Zn^{2+}$, $Ni^{2+}$, and $Ca^{2+}$ reduced the enzyme activity slightly. When treated with the B. megaterium VHM1 enzyme, the flatulence-causing sugars in soymilk were completely hydrolyzed within 1.5 h.

• Microbiology and Biotechnology Letters
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• v.16 no.6
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• pp.499-504
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• 1988

Among the new yeast strains which were isolated from soils by incubating in the xylan containing minimal medium at 3$0^{\circ}C$, one strain(XB-33) was finally selected by the results of extracellular xylanase production test. The characteristics of XB-33 was almost consistent with those of the Cryptococcus ater. The formation of xylanase activity was induced by xylan and repressed by xylose or glucose. The xylanase was partially purified from the culture supernatant with DEAE-Sephadex A5O chromatography. The enzyme had a pH optimum for activity at 5.0 and its stability range was pH 5-7. The temperature optimum was at 5$0^{\circ}C$, but the enzyme activity was greatly lost by heating at 7$0^{\circ}C$ for 60 minutes. The hydrolysis products from xylan by crude enzyme detected by TLC, were xylose and n series of higher oligosaccharides. The Km value of xylanase was 20 (mg/ml).

• Journal of Ginseng Research
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• v.45 no.1
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• pp.48-57
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• 2021

Background: Ginsenoside Rh2 is well known for many pharmacological activities, such as anticancer, antidiabetes, antiinflammatory, and antiobesity properties. Glycosyltransferases (GTs) are ubiquitous enzymes present in nature and are widely used for the synthesis of oligosaccharides, polysaccharides, glycoconjugates, and novel derivatives. We aimed to synthesize new ginsenosides from Rh2 using the recombinant GT enzyme and investigate its cytotoxicity with diverse cell lines. Methods: We have used a GT gene with 1,224-bp gene sequence cloned from Lactobacillus rhamnosus (LRGT) and then expressed in Escherichia coli BL21 (DE3). The recombinant GT protein was purified and demonstrated to transform Rh2 into two novel ginsenosides, and they were characterized by nuclear magnetic resonance (NMR) techniques and evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay. Results: Two novel ginsenosides with an additional glucopyranosyl (6→1) and two additional glucopyranosyl (6→1) linked with the C-3 position of the substrate Rh2 were synthesized, respectively. Cell viability assay in the lung cancer (A549) cell line showed that glucosyl ginsenoside Rh2 inhibited cell viability more potently than ginsenoside Rg3 and Rh2 at a concentration of 10 μM. Furthermore, glucosyl ginsenoside Rh2 did not exhibit any cytotoxic effect in murine macrophage cells (RAW264.7), mouse embryo fibroblasts cells (3T3-L1), and skin cells (B16BL6) at a concentration of 10 μM compared with ginsenoside Rh2 and Rg3. Conclusion: This is the first report on the synthesis of two novel ginsenosides, namely, glucosyl ginsenoside Rh2 and diglucosyl ginsenoside Rh2 from Rh2 by using recombinant GT isolated from L. rhamnosus. Moreover, diglucosyl ginsenoside Rh2 might be a new candidate for treatment of inflammation, obesity, and skin whiting, and especially for anticancer.

• Journal of the Korean Applied Science and Technology
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• v.32 no.4
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• pp.758-766
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• 2015

Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.