• Clinical and Experimental Pediatrics
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• v.52 no.10
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• pp.1061-1068
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• 2009

The treatment of pediatric patients with chronic renal disease comprises management of nutritional imbalance, fluid, electrolyte, and acid-base disturbances, mineral bone disease, anemia, hypertension, and growth retardation. The treatment also includes administration of appropriate renal replacement therapy, if required. Adequate dietary intake of carbohydrates, fats, and proteins and caloric intake must be encouraged in such patients to ensure proper growth and development. In addition, fluid, electrolyte, and acid-base status must be regularly monitored and should be well maintained. Serum calcium, phosphorus, and parathyroid hormone levels must be maintained at their target range, which are determined on the basis of the glomerular filtration rate, to avoid the development of mineral bone disease. This can be achieved by using phosphorus binders and vitamin D analogues. An erythropoiesis-stimulating agent must be administered along with iron supplementation to maintain the hemoglobin level of the patients between 11-12 g/dL. Hypertension must be controlled with adequate water and sodium balance and appropriate antihypertensive agents. Administration of recombinant human growth hormone is recommended to improve the final adult heights.

• Korean Journal of Microbiology
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• v.41 no.2
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• pp.93-98
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• 2005

The sphingosine kinase gene, which is 969-nucleotide long, was identified during the whole genome sequencing of Sphingomonas chungbukensis DJ77. The amino acid sequence showed the identity of $55\%$ with that of Zymomonas mobilis subsp. mobilis ZM4. C2, C3, and C5 domains of eukaryotic sphingosine kinase were found in sphingosine kinase from Sphingomonas chungbukensis DI77. One of these three conserved sites, GGDG, was predicted as a ATP-binding site, and the functions of the others were unknown currently. The phylogenetic tree constructed by ClustalX indicated that the sphingosine kinase of S. chungbukensis DJ77 was near the phylogenetic group COG1597, and did not belong to the group of diacylglycerol kinase of the same strain. The recombinant sphingosine kinase was expressed in Escherichia coli, but it was made in form of inclusion body.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.141-146
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• 2012

A gene encoding the xylanase (XynA) predicted from partial genomic sequence of Paenibacillus woosongensis was cloned into Escherichia coli by PCR. This xynA gene consisted of 633 nucleotides, encoding a polypeptide of 211 amino acid residues. The deduced amino acid sequence exhibited 85-89% identity with those of several Paenibacillus xylanases, belonging to the glycosyl hydrolase family 11. As a results of expression of the structural gene by T7 promoter of a pET23a(+) expression vector, xylanase activity was higher in cell-free extract than culture filtrate of a recombinant Escherichia coli BL21(DE3) CodonPlus. However, the expression level of xylanase was not sufficient be detected by SDS-PAGE. The cell-free extract showed maximal xylanase activity at $60^{\circ}C$ and pH 5.5. The predominant products resulting from xylan and xylooligosaccharide hydrolysis were xylose and xylotriose. The enzyme could hydrolyze xylooligosaccharides larger than xylbiose.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.241-246
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• 2002

A gene coding for endo-$\beta$-1,3-1,4-glucanase of Bacillus circulans was subcloned into Escherichia coli Ml5 using pQE30 as an expression vector. Endo-$\beta$-1,3-1,4-glucanase produced by the recombinant expression plas-mid pLQ43 was intactly purified to a single protein through a nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography method. The molecular mass of the purified enzyme was estimated to be 28 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme activity were pH 6.8 and $60^{\circ}C$, respectively. This enzyme was fairly stable in the pH ranging 5.5~7.5 and at the temperatures lower than $55^{\circ}C$. The enzyme appeared to be sensitive to most of the metal ions, especially to $Hg^{2＋$, and also to methanol, ethanol, isopropanol or 1-butanol at a concentration of 10%(v/v).

• Journal of Applied Biological Chemistry
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• v.62 no.4
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• pp.361-366
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• 2019

Isorhamnetin 3-O-glucoside, a member of the flavonol group, has been reported to be effective for inflammatory and ulcer, as well as to alleviate diabetic complications such as neuropathy, nephropathy and retinopathy. Isorhamnetin 3-O-glucoside has been extracted from several plants. Biotransformation is a valuable tool, which is used to produce value-added chemicals with inexpensive compounds. To synthesis isorhamnetin 3-O-glucoside from quercetin, two genes (PGT E82L and ROMT-9) were introduced into Escherichia coli, respectively. In order to synthesis isorhamnetin 3-O-glucoside from quercetin, a co-culture fermentation system was developed by optimizing the medium and temperature for biotransformation, the cell mix ratio, Isopropyl-β-ᴅ-thiogalactoside induction time, and quercetin feed concentration. Finally, isorhamnetin 3-O-glucoside was biosynthesized up to 181.2 mg/L under the optimized biotransformation condition, which was higher 4.7 times than previously reported (39.6 mg/L).

• Journal of Food Hygiene and Safety
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• v.35 no.4
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• pp.304-311
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• 2020

Galacto-oligosaccharides (GOS) are prebiotics that have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut, in addition to having various applications in the food industry. GOS are generally produced from lactose in a reaction catalyzed by β-galactosidase. Synthesis of GOS from whey permeate (WP) (ultrafiltration of whey, concentrated then spray dried) using surface engineered β-galactosidase in Pichia pastoris (P. pastoris) is a novel method to convert waste into a valuable product. Cell-surface display is the expression of peptides and proteins on the surface of living cells by fusing them to functional components of cells. Surface engineered cells have many potential uses. The Flo1p flocculation functional domain, thought to be located near the N terminus, recognizes and adheres non-covalently to cell-wall components such as α-mannan carbohydrates, causing reversible aggregation of cells into flocs.

• Journal of Life Science
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• v.8 no.2
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• pp.181-188
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• 1998

Two distinct ADP-ribosyltransferases, termed Yac-1 and Yac-2 from mouse lymphoma cells were recently cloned and characterized. Yac-1 enzyme possesses ADP-ribosyltransferases activity. In contrast, Yac-2 has significant NAD glycohydrolase activity and may preferentially hydrolyze NAD. Yac-2 possesses a glutamate at position 219 adjacent to the two consdrved glutamic acid residues. To study the effect of Glu-219 on enzyme activities, Glu-219 was mutagenized to Glutamine (E219Q) or alanine (E219A) using a two-step recombinant polymerase chain reaction procedure. Replacing Glu at position 219 with Gln or Ala resulted in 56 (E219Q) or 66% (E219A) reduction in ADP-ribosyltranferase activity. The NAD glycohydrolase activity of Yac-2 protein were not altered by the mutations. These results indicate that Glu-219 in Yac-2 enzyme plays an important role in ADP-ribosyltransferase, but not NAD glycohydrolase activity.

• Journal of Life Science
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• v.20 no.11
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• pp.1582-1588
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• 2010

A new alginate lyase gene of marine bacterium Streptomyces sp. M3 had been previously cloned in pColdI vector and transformed into E. coli BL21 (DE3). In this study, M3 lyase protein without signal peptide was overexpressed by induction with IPTG and purified with Ni-Sepharose affinity chromatography. The absorbance at 235 nm of the reaction mixture and TLC analysis showed that M3 alginate lyase was a polyG-specific lyase. When M3 lyase was assayed with substrate for 10 min, optimum pH and optimum temperature were pH 9 and $60^{\circ}C$. For the effect of 1mM metal ion on M3 lyase activity, $Ca^{++}$ and $Mn^{++}$ ions increased the alginate degrading activity by two-fold, whereas $Hg^{++}$ and $Zn^{++}$ ions inhibited the lyase activity completely. $Mg^{++}$, $Co^{++}$, $Na^+$, $K^+$, and $Ba^{++}$ did not show any strong effects on alginate lyase activity.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.207-212
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• 2010

A bacterium producing the extracellular mannanase was isolated from Korean soybean paste. The isolate WL-8 has been identified as Bacillus subtilis on the basis on its 16S rRNA sequence, morphology and biochemical properties. The mannanase productivity of strain WL-8 was increased in LB broth by addition of wheat bran. The maximum mannanase productivity was reached to approximately 20 U/ml in LB medium supplemented with 6% wheat bran. A gene encoding the mannanase of WL-8 was cloned into Escherichia coli and its nucleotide sequence was subsequently determined. The mannanase gene consisted of 1,086 nucleotides encoding a polypeptide of 362 amino acid residues. The deduced amino acid sequence was highly homologous with those of several mannanases from B. subtilis belonging to GH family 26. Reaction temperature and pH profiles were investigated using the culture filtrate and cell-free extract of the recombinant E. coli carrying a WL-8 mannanase gene, respectively. Optimal conditions for the two fractions occurred at pH 5.5 and $60^{\circ}C$. The cell-free extract showed higher mannanase activity than the culture filtrate at above $60^{\circ}C$.

• Korean Journal of Veterinary Research
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• v.46 no.2
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• pp.127-133
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• 2006

Pasteurella multocida is a terrible veterinary pathogen that causes widespread infections in husbandry. To induce homologous and/or heterologous immunity against the infections, outer membrane protein Hs (OmpH) in the envelope of different strains of P. multocida are thought to be attractive vaccine candidates. Previously we cloned and characterized a gene for OmpH from pathogenic P. multocida (A : 3) (In Press, Korean J. Microbiol. Biotechnol. 2005, 33, December). The gene is composed of 1,047 nucleotides (nt) coding 348 amino acids (aa) with signal peptide of 20 aa. The truncated ompH, a gene without nt coding for the signal peptide, was generated using pRSET A to name "pRSET A/OmpH-F2". This truncated ompH was well expressed in Escherichia coli BL21 (DE3). Truncated OmpH was purified for induction of immunity against live pathogen of fowl cholera (P. multocida A : 3) in mice. Some $50{\mu}g$ of the purified polypeptide was intraperitoneally injected into mice two times with 10 day interval. Lethal dose ($25{\mu}l$) of live P. multocida A : 3 was determined by directly injecting the pathogen into wild mice (n = 25). To demonstrate the vaccine candidate of the truncated OmpH, the live pathogen ($25{\mu}l$) was challenged with the OmpH-immunized mouse group as well as positive & negative controls (n = 80). The results show that the truncated OmpH can be used for an effective vaccine production to prevent fowl cholera caused by pathogenic P. multocida (A : 3).