• Microbiology and Biotechnology Letters
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• v.47 no.1
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• pp.78-86
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• 2019

Recombinant Corynebacterium glutamicum producing N-acetylglucosamine (GlcNAc) was constructed by metabolic engineering. To construct a basal strain producing GlcNAc, the genes nagA, nagB, and nanE encoding N-acetylglucosamine-6-phosphate deacetylase, glucosamine-6-phosphate deaminase, and N-acetylmannosamine-6-phosphate epimerase, respectively, were sequentially deleted from C. glutamicum ATCC 13032, yielding strain KG208. In addition, the genes glmS and gna1 encoding glucosamine-6-phosphate synthase and glucosamine-6-phosphate N-acetyltransferase, which originated from C. glutamicum and Saccharomyces cerevisiae, respectively, were expressed in several expression vectors. Among several combinations of glmS and gna1 expression, recombinant cells expressing glmS and gna1 under control of the ilvC promoter produced 1.77 g/l of GlcNAc and 0.63 g/l of glucosamine in flask cultures.

• Development and Reproduction
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• v.4 no.2
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• pp.181-186
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• 2000

The expression of glucosamine-6-phosphate deaminase (GNPDA) was examined in mouse ovary from neonate to aduit. In western blot, band of Mr. 31 kDa antigen sharply increased 2 weeks after birth onward. In irmmunostaining of the adult ovary, GNPDA expression was constitutive in the theca and interstitial cells. However, expression in the granulosa cells was different according to folliculogenesis. Cytoplasm of the oocyte of some primary follicle showed positive signal but not in the antral follicle. Granulosa cells of antral follicles showed no visible sign of GNPDA expression. In the corpora lutea, the signal intensity in granulosaluteal cells increased according to luteal development and became the highest in the luteolytic phase. In summary, the differential expression of GNPDA was found in follicle cells according to folliculogenesis. It suggests that GNPDA might be involved in tissue remodeling in mouse ovary.

• Journal of Nutrition and Health
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• v.47 no.1
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• pp.1-11
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• 2014

Purpose: Several studies have proven that EGCG, the primary green tea catechin, and glucosamine-6-phosphate (PGlc) reduce triglyceride contents in 3T3-L1 adipocytes. The objective of this study is to evaluate the combination effect of EGCG and PGlc on decline of accumulated fat in differentiated 3T3-L1 adipocytes. Methods: EGCG and PGlc were administered for 6 day for differentiation of 3T3-L1 adipocytes. Cell viability was measured using the CCK assay kit. In addition, TG accumulation in culture 3T3-L1 adipocytes was investigated by Oil Red O staining. We examined the expres-sion level of several genes and proteins associated with adipogenesis and lipolysis using real-time RT-PCR and Western blot analysis. A flow cytometer Calibar was used to assess the effect of EGCG and PGluco on cell-cycle progression of differentiating 3T3-L1 cells. Results: Intracelluar lipid accumulation was significantly decreased by combination treatment with EGCG $60{\mu}M$ and PGlc $200{\mu}g/m$ compared with control and EGCG treatment alone. In addition, use of combination treatment resulted in directly decreased expression of $PPAR{\gamma}$, $C/EBP{\alpha}$, and SREBP1. In addition, it inhibited adipocyte differentiation and adipogenesis through downstream regulation of adipogenic target genes such as FAS, ACSL1, and LPL, and the inhibitory action of EGCG and PGlc was found to inhibit the mitotic clonal expansion (MCE) process as evidenced by impaired cell cycle entry into S phase and the S to G2/M phase transition of confluent cells and levels of cell cycle regulating proteins such as cyclin A and CDK2. Conclusion: Combination treatment of EGCG and PGlc inhibited adipocyte differentiation through decreased expression of genes related to adipogenesis and adipogenic and cell cycle arrest in early stage of adipocyte differentiation.

• KSBB Journal
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• v.18 no.3
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• pp.180-185
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• 2003

Glycosaminoglycans (GAGS was extracted from sea squirt, Styela clava with sodium phosphate at 105$^{\circ}C$ for 2 hr and deproteinized with trichloroacetic acid or hydrochloride. The GAGs obtained from tunic consist 41.7% crude carbohydrates, 31.8% crude protein, and 31.2% sulfate. It was mainly constituted of galactose, glucosamine, glucose, mannose, and glacrosamine. The prominent amino acid were phenylalanine, threonine, glutamic acid, and aspartic acid. Mineral contents was mainly constituted 3.0 mg% sodium, 1.6 mg% potassium, and 1.2 mg% phosphorus. Trichloroacetic acid, hydrochloride and 5-sulfosalicylic acid were used for deprotein of the GAGs. Effective volume for deprotein of crude GAGs were 5.0% trichloroacetic acid (w/v) and 10.0% HCI (v/v) treatment. The deproteinized GAGs contained 35.1%, 35.4% of protein and 22.0%, 18.5% of sulfate, respectively.

• Journal of the Korean Wood Science and Technology
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• v.28 no.1
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• pp.55-64
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• 2000

Culture maturity assessment can be used to control fruiting body flush timing. Culture maturity of sawdust-based substrate was evaluated by using oak mushroom, (Lentinula edodes (Berk.) Pegler). The influence of substrate water potential (${\psi}$) on the growth and fruiting of three genotypes of L. edodes was also investigated. Glucosamine content revealed a peak at the fruiting body senescent stage. Glucosamine increased steadily to the sporophore senescent stage, and sharply declined at crop treatment. Lipid phosphate and ergosterol contents peaked at pinning and button break stages, respectively. Therefore lipid phosphate and ergosterol contents would be considered as the convenient measurement for judging culture maturity and fruiting potentials. The substrate pH values before inoculation and on the fruiting stage were varied from 6.3 to 4.0. This pH changes were detected as changes in color from bluish purple to yellow by direct bromphenol blue(BPB) spraying, and shown a good correlation with fruit body yield of the 1 st flush. Concerning water potential of the cultures, a slight reduction of water potential, -0.5MPa, stimulated mycelial and colony growths on liquid, agar and sawdust-based substrates. The water potential of well-colonized matured substrate was -0.7MPa and -4.0MPa, before and after the fruiting, respectively. Excellent water providing capacity (higher ${\psi}$) is expected to well-matured cultures with a high density of mycelial colonization. Also, the substrate water potential significantly affected by the interaction between genotypes and spawn run time.

• The Journal of the Korean Society for Microbiology
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• v.20 no.1
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• pp.55-63
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• 1985

Phosphate, ammonia, glucosamine, glucose, pyruvate, succinate, fumarate, malate and acetate were examined for their ability to control the heat-stable enterotoxin (ST) production in succinate salts medium or in M9 medium. The results obtained were summerized as follows. 1. When the initial phosphate concentration was adjusted to 1.0mM, ST production was decreased to 80u/ml or less. But when the initial phosphate concentration was adjusted to 64mM or 100mM, enterotoxin production was 320u/ml. 2. When the initial ammonia concentration in the medium was adjusted to 1.0mM, no ST production and cell growth were observed. But when ammonia concentration was adjusted to 10mM, 19mM, 38mM or 76mM, enterotoxin production was 320u/ml. 3. Among carbon sources, glucosamine, glucose, pyruvate, succinate, fumarate, malate and acetate, acetate supported the highest specific production (928 unit/O.D.) of heat-stable enterotoxin. From this results, we could assume that heat-stable enterotoxin production is controlled by stringent control mechanism. 4. When the pH of the succinate salts medium was kept between 6.2 to 6.5, no heat-stable enterotoxin production was observed, but when the pH of the medium was kept between pH 6.2 to 6.5, 267 unit/O.D. of heat-stable enterotoxin was produced. 5. Glucose inhibited the heat-stable enterotoxin production and the mechanism was assumed due to its capacity to lower the pH of the medium during catabolysis and its high metabolic energy.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.6
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• pp.442-446
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• 2007

From the rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass, unsoluble inorganic phosphate solubilizing bacterial strains were isolated using plate base assay on Pikovskaya's medium. Two strains, CL-1 and CL-2, which produced largest halo on plates (indicative of phosphate solubilization)were selected for further studies. Based on these biochemical and 16S rRNA analysis strains CL-1, CL-2 were found to be as species of Pseudomonas sp. and Kluyvera sp., respectively. In broth assay Pseudomonas sp. CL-1 and Kluyvera sp. CL-2 solubilized insoluble phosphate by 193.4 mg and $493.6P\;mg\;L^{-1}$, respectively after $3^{rd}$ day inoculation. These effecient phosphate solubilizing bacteria have a potential to be developed as microbial based fertilizer in future.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.475-479
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• 2003

Glycosaminoglycans(GAGs) from sea squirt, Styela clava was extracted with sodium phosphate at $105^{\circ}C$ for 2 hr and deprotein with trichloroacetic acid or hydrochloride. This GAGs was mainly constituted of galactose, glucosamine, glucose, mannose and galactosamine, and was phenylalanine, threonine, glutamic acid and aspartic acid. Mineral contents was mainly constituted 3.0mg% sodium, 1.6mg% potassium and 1.2mg% phosphorus and heavy metal was not detected. At pharmaceutical and cosmetic code of GAGs, protein and sulfate contents should included each range $14.0{\sim}22.0%$, $35.0 {\sim}45.0%$. After 5.0% trichloroacetic acid(w/v) and 10.0% HCl(v/v) treatment, protein and sulfate contents of GAGs was contained each 35.1%, 35.4% and each 22.0%, 18.5%.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.227-232
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• 2015

A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

• Microbiology and Biotechnology Letters
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• v.5 no.4
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• pp.177-184
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• 1977

Arylsulfatase catalyzes the release of SO$\sub$4//sup2/- from sulfate esters of simple phenols. Arylsolfatase occurs widely in animal tissues and in microorganisms including soil bacteria. Its widespread distribution suggests that it has a rather fundamental function and environmental meaning. It has been shown previously that arylsulfatase of Klebsiella was purified and characterized. A condition of arylsulfatase synthesis was tested with several strains of Serratia. Serratia marcescens could not utilize some sugars, such as xylose, rhamnose, glucosamine and arabinose hut glucose and mannitol as a sole carbon source. However, arylsulfatase synthesis was repressed by glucose but not by mannitol. The enzyme synthesis was repressed ob inorganic sulfate and methionine, and this repression was relieved by addition of tyramine. Arylsulfatase of S. marcescen was purified by fractionation with ammonium sulfate and followed by chromatographies on DEAE-Cellulose CM-Cellulose, and DEAE-Sephadex A-25. The molecular weight of arylsulfatase was determined to be 46,000 by SDS-Gel electrophoresis and 49,000 by Sephadex G-100 column chromatography. The enzyme showed some different properties with that of K. aerogenes. The activity was maximum at pH 6.8. The Km and Vmax values for p-nitrophenyl sulfate were 2.5${\times}$10$\^$－4/ M and 20 nmoles/min/mg protein, respectively. The enzyme showed high activities toward phenyl sulfate, ο-and p-nitro phenyl sulfates, and p-nitrocatechol sulfate. The inhibition of enzyme was strongly affected by hydroxylamine, inorganic fluoride, sulfide and phosphate, but by inorganic sulfate. Like Klebsiella arylsulfatase, tyramine, octopamine, and dopamine gave signifcant inhibitory effect.