• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1534-1538
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• 2010

Fed-batch cultures of Hansenula polymorpha were studied to develop an efficient biosystem to produce recombinant human serum albumin (HSA). To comply with this purpose, we used a high-purity oxygen-supplying strategy to increase the viable cell density in a bioreactor and enhance the production of target protein. A mutant strain, H. polymorpha GOT7, was utilized in this study as a host strain in both 5-l and 30-l scale fermentors. To supply high-purity oxygen into a bioreactor, nearly 100% high-purity oxygen from a commercial bomb or higher than 93% oxygen available in situ from a pressure swing adsorption (PSA) oxygen generator was employed. Under the optimal fermentation of H. polymorpha with highpurity oxygen, the final cell densities and produced HSA concentrations were 24.6 g/l and 5.1 g/l in the 5-l fermentor, and 24.8 g/l and 4.5 g/l in the 30-l fermentor, respectively. These were about 2-10 times higher than those obtained in air-based fed-batch fermentations. The discrepancies between the 5-l and 30-l fermentors with air supply were presumably due to the higher contribution of surface aeration over submerged aeration in the 5-l fermentor. This study, therefore, proved the positive effect of high-purity oxygen in enhancing viable cell density as well as target recombinant protein production in microbial fermentations.

• ALGAE
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• v.30 no.2
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• pp.147-161
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• 2015

Brown algal extracts have long been used as feed supplements to promote health of farm animals. Here, we show new molecular insights in to the mechanism of action of a fucose containing polymer (FCP) rich fraction from the brown seaweed Ascophyllum nodosum using the Caenorhabditis elegans-Pseudomonas aeruginosa PA14 infection model. FCP enhanced survival of C. elegans against pathogen stress, correlated with up-regulation of key immune response genes such as: lipases, lysozyme (lys-1), saponin-like protein (spp-1), thaumatin-like protein (tlp-1), matridin SK domain protein (msk-1), antibacterial protein (abf-1), and lectin family protein (lfp). Further, FCP caused down regulation of P. aeruginosa quorum sensing genes: (lasI, lasR, rhlI, and rhlR), secreted virulence factors (lipase, proteases, and elastases) and toxic metabolites (pyocyanin, hydrogen cyanide, and siderophore). Biofilm formation and motility of pathogenic bacteria were also greatly attenuated when the culture media were treated with FCP. Interestingly, FCP failed to mitigate the pathogen stress in skn-1, daf-2, and pmk-1 mutants of C. elegans. This indicated that, FCP treatment acted on the regulation of fundamental innate immune pathways, which are conserved across the majority of organisms including humans. This study suggests the possible use of FCP, a seaweed component, as a functional food source for healthy living.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.5
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• pp.566-572
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• 1998

The present study examined the effects of excessive dietary protein and energy on growth response to clenbuterol in broilers. The chicks were allocated into 6 groups at 14d old, and used for a $3{\times}2$ factorial experiment. Birds were fed six diets, the control diet containing 21% crude protein (CP) and 3,100 kcal of metabolizable energy ME/kg, a high protein (30% CP) or a high energy (3,500 kcal/ ME/kg) diet, with or without 1 ppm clenbuterol, for 18 d. Clenbuterol feeding markedly decreased (p < 0.05) body weight gain by 23% in the high energy group. Feed intake was also decreased (p < 0.05) by clenbuterol administration across diet treatments. Abdominal fat weight was reduced (p < 0.05) by clenbuterol only when chickens were fed the high energy diet. Clenbuterol increased (p < 0.05) leg muscle weight in the control diet group, but decreased (p < 0.05) it in the high energy group. Muscle protein concentration was increased by 11 % in leg muscle only of the birds at the high energy level. In leg muscle, clenbuterol enhanced the protein/DNA ratio by 18%, except for the high protein group. These results indicate that feeding a diet containing excessive amounts of protein and more energy than normal did not necessarily improve growth response to clenbuterol.

• Journal of Microbiology
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• v.39 no.1
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• pp.24-30
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• 2001

In addition to effecting the catalysis of sugar uptake, the bacterial phosphoenolpyruvate::sugar phosphotransferase system regulates a variety of physiological processes. In a previous paper [Seok et al.,(1997) J. Biol. Chem. 272, 26511-26521], we reported the interaction with and allosteric regulation of Esiherichia coli glycogen phosphorylase activity by the histidine-containing phosphocarrier protein HPr in vitro. Here, we show that the specific interaction between HPr and glycogen phosphorylase occurs in vivo. To address the physiological role of the HPr-glycogen phosphorylase complex, intracellular glycogen levels were measured in E. coli strains transformed with various plasmids. While glycogen accumulated during the transition between exponential and stationary growth phases in wildtype cells, it did not accumulate in cells overproducing HPr or its inactive mutant regardless of the growth stage. From these results, we conclude that HPr mediates crosstalk between sugar uptake through the phosphoenolpyruvate:sugar phosphotransferase system and glycogen breakdown. The evolutionary significance of the HPr-glycogen phosphorylase complex is suggested.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.2
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• pp.148-153
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• 2006

Effects of ${\alpha}$-chymotrypsin modification on degree of hydrolysis (DH), solubility, emulsifying capacity and thermal aggregation of laboratory-purified soy protein isolate (SPI) using a lipoxygenase-defected soybean (Jinpum-kong) and commercial soy protein isolate (Supro 500E) were compared. SPIs were hydrolyzed by ${\alpha}$-chymotrypsin at pH 7.8 and $37^{\circ}C$ for 30 min. DHs of Supro 500E and Jinpum-kong SPI were increased by ${\alpha}$-chymotrypsin modification, and DH of Supro 500E was higher than that of Jinpum-kong SPI. DH of ${\alpha}$-chymotrypsin treated Jinpum-kong SPI was similar with untreated Supro 500E and DH of treated Supro 500E was the highest. Solubility, emulsifying capacity and thermal aggregation of SPIs were increased by ${\alpha}$-chymotrypsin modification, and these changes were highly related to changes in DH. Functional properties of Supro 500E were higher than Jinpum-kong SPI in both of untreated and ${\alpha}$-chymotrypsin treated SPIs.

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.115-118
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• 2006

Cyclin B1 is known to reflect the M-phase promoting factor (MPF), a universal regulator of G2/M-phase transition, activity during the process of oocytes maturation. To investigate whether culture condition affects the maturation rate and the expression of cyclin B1 protein, bovine immature oocytes are stimulated and cultured according to the following protocols: Experiment 1: denuded oocytes (denude) only, COC only, denuded oocytes+granulosa cells (denude+GCs) and COC+GCs; Experiment 2: no-activation (control), 7% ethanol for 5 min and $10{\mu}l/ml$ ionomycin for 5 min at immediately before maturation. The maturation rates of denude and no-activation group were significantly lower in both experiments (P<0.05), respectively. Co-culture or stimulation method in bovine immature oocytes culture increases the cyclin B1 expression significantly in both experiments (P<0.05). Based on these results, culture condition affects the maturation rate and the expression of cyclin B1 protein during the first meiotic maturation in bovine immature oocytes.

• Korean Journal of Veterinary Service
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• v.30 no.2
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• pp.205-209
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• 2007

A protein chip based on surface plasmon resonance (SPR) imaging was developed for measuring classical swine fever virus (CSFV) antibody using a recombinant gp55 protein as an antigen. The diagnostic potential of SPR imaging for detecting antibodies to the CSFV gp55 protein was compared with that of a enzyme -linked immunosorbent assay (ELISA) using 70 pig sera. There was a strong positive correlation between the SPR imaging and ELISA (n=70, r=0.916, p<0.01). Therefore, the SPR imaging, which is a label-free and high-through put method, is expected to be a valuable tool in the serodiagnosis of CSFV.

• BMB Reports
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• v.30 no.3
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• pp.222-228
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• 1997

Protein phosphatase 2C (PP2C) is one of the four major serine/threonine phosphatases which is dependent on $Mg^{2+}$ for its activity. PP2C was purified from rat liver cytosol and its characteristics were investigated. The substrate employed for routine assay was $[^{32}P]casein$ phosphorylated by PKA. The purification process involved DEAE chromatography, ammonium sulfate fractionation, phenyl sepharose chromatography, sephacryl 5-200 gel filtration, and histone agarose chromatography. The SDS-PAGE of PP2C showed one major single protein band at a position corresponding to a molecular mass of 43 kd and the purification fold was 637. The enzyme showed a pH optimum of 8 and $K_M$ value was $1.9\;{\mu}M$. However, when the substrate was changed to $[^{32}P]histone$, the pH optimum was shifted to 7 and $K_M$ value was $2.3\;{\mu}M.\;Mg^{2+}$ was essential to the enzyme activity and okadaic acid did not exert any inhibitory effect on the enzyme. To examine residue in the active site of PP2C effects of some protein-modifying reagents were tested.

• Journal of the Korean Wood Science and Technology
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• v.46 no.6
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• pp.670-680
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• 2018

Acanthophysium sp. KMF001, a wood rotting fungus, produces a strong crude enzyme complex that efficiently produces simple sugars from wood. The transcriptomic analysis of Acanthophysium sp. KMF001 identified 14 genes for putative glycoside hydrolases. Among them, isotig01043 was expressed heterogeneously in Escherichia coli BL21(DE3), and the expressed protein exhibited an endo-${\beta}$-1,4-xylanase activity which showed the optimum reaction at pH 5.0 and $30^{\circ}C$. The enzyme kinetic values of $K_m$ and $V_{max}$ were 25.92 mg/ml and $0.628{\mu}mole/mg/ml$, respectively. The enzymatic characteristics of the expressed xylanase showed a typical fungal xylanase. However, the bioinformatics analysis suggested that the protein encoded by isotig01043 was a novel xylanase based on a low identity when it was compared with the closest protein in the NCBI database and a similar protein domain with GH16_fungal_Lam16A_glucanase, which had not been earlier suggested as a xylanase.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.11
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• pp.1548-1553
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• 2000

An experiment was conducted to study the nutritional quality of whole crop corn silage ensiled with cage layer manure in sheep. Treatments were designed as a $3{\times}3$ Latin square with 16-day periods. Sheep were allotted in one of three diet-treatments, which were whole crop corn silage (CS), whole crop corn+30% cage layer manure (CLM) silage (based on DM; MS) and rice straw+concentrate (SC) mixed at 8:2 ratio (on DM basis). Silage ensiled with CLM significantly increased (p<0.05) digestibilities of crude protein, NDF and ADF, TDN over the other treatments. Ruminal pH in sheep fed SC was significantly (p<0.05) higher than that of the other diets at 0.5, 1, 2, 4 and 8 h after feeding. Ruminal ammonia nitrogen concentration of the MS treatment was significantly (p<0.05) higher than that of the other treatments at 0, 1, 2 h after feeding. The MS treatment highly increased (p<0.05) feed intake, digestibility of organic matter and crude protein, nitrogen intake and retained nitrogen. The MS treatment highly increased (p<0.05) purine derivative (PD) excretion leading to higher microbial protein synthesis.