• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.749-754
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• 2000

PCR primers were designed based on consensus sequences of dTDP-D-glucose 4,6-dehydratase, one of the enzymes involved in the biosynthesis of deoxysugar. The PCR product (360 bp) was obtained from Thermus caldophilus GK24. Colony hybridization was carried out to the cosmid library constructed from T. caldophilus GK24 genomic DNA by the PCR product DNA fragment. We isolated a cosmid clone (pSMTC-1) that was subcloned to call pKCB series plasmid (BamHI fragments), partially sequenced and analyzed. pKCB80 (4.2 kb-BamHI DNA fragment) of them showed ORFs that was orfA, orfB, orfC and orfD. The orfABCD gene cluster is the deosysugar biosynthetic gene ; orfA (glucose-1-phosphate thymidylytransferase), orfB (dTDP-D-glucose 4,6-dehydratase), orfC (dTDP-4-keto-L-rhamnose reductase) and orfD (dTDP-4-keto-6-deoxy-D-glucose 3,5-epimerase). The gene cluster that was related in biosynthesis of dTDP-L-rhamnose was also identified by computer analysis, and we proposed that the biosynthetic pathway of deoxysugar analyzed from DNA sequencing of pKCB80 is from D-glucose-1-phosphate, dTDP-D-glucose, dTDP-4-keto-6-deoxy-D-glucose via dTDP-4-keto-L-rhamnose to dTDP-L-rhamnose.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.206-212
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• 1999

The gene orf7(oxi III) was expressed using an E. coli system in anticipation that it would encode dTDP-glucose 4,6-dehydratase which is involved in the biosynthesis of the olivose moiety of chlorothricin produced from Streptomyces antibioticus Tu99. The solubility of the expressed protein increased up to 20% under optimal induction conditions. The expressed protein was purified from the E. coli BL 21(DE3) cell lysate by a 28.5-fold purification in two chromatography steps with a 38% recovery to near homogeneity. The molecular weight and N-terminal amino acid sequence of the purified protein correlated with the predicted mass and sequence deduced from the orf7 gene. The purified protein was a homodimer with a subunit relative molecular weight of 38,000 Dalton. The expressed protein was found to exhibit dTDP-glucose 4,6-dehydratase activity and be highly specific for dTDP-glucose as a substrate. The values of K'm and V'max for dTDP-glucose were 28 $\mu$M and 295 nmol $min^{-1} (mg protein)^{-1}$, respectively. dTTP and dTDP were strong inhibitors of this enzyme.$NAD^+$, the coenzyme for dTDP-glucose 4,6-dehydratase, was tightly bound to the expressed protein.

• Biomedical Science Letters
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• v.11 no.2
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• pp.243-248
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• 2005

The present study was conducted to determine whether skin spread of Myrrha and Tending Diancibo Pu (TDP) irradiation have a remarkable effect on the cell regeneration as well as wound healing following dermal scald burn injury. Burn injury was induced on dorsal surface $(TBSA\;15\~20\%)$ by scald burn in rats. Postburn concentration of serum protein was significantly decreased compared with sham-treated, double treatment with Myrrha and TDP was significantly increased the protein concentration compared with that of burn control. The content of keratinocyte growth factor (KGF) at 48 h is higher than that of at 24 h, and double treatment with Myrrha and TDP was the most effective to increase the production of KGF in all experimental groups. Morphologically, epithelial regeneration and dermal collagen synthesis by fibroblasts were accelerated in Myrrha and TDP treated group compared with bum control at same time postburn. At 48 h after burn, all dermal connective tissues are recovered to new collagen fibers in case of Myrrha and TDP double treated group. The data suggest that double treatment with skin spread of Myrrha and TDP radiation have a remarkable effect of to accelerate cell regeneration and wound healing in case of scald burn skin.

• Proceedings of the Korean Society for Emotion and Sensibility Conference
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• 2009.05a
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• pp.67-70
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• 2009

본 논문에서는 생리신호를 이용한 감성인식의 정확도를 높이기 위한 개인화 방안에 대해 연구하였다. 이전연구 결과인 TDP(Time Dependent Parameters)를 이용한 감성인식방법은 자율신경계 반응을 4 단계로 세분화하여 감성인식의 정확도를 높일 수 있었다. 하지만 평균 정확도는 향상된 반면 개인별로 정확도의 개인차가 발생하였다. 본 연구는 개인차를 줄이기 위해서, 개인의 반응에 따라 감성인식을 위한 룰베이스가 변화하는 적응적 TDP 알고리즘을 개발하였다. 시각자극을 이용한 감성유발 실험결과를 분석하여 감성인식 개인차가 감소하였는지 확인하였다. 실험은 4 명을 대상으로 하였으며 한 명당 24번의 시각 자극을 제시하여 96개의 데이터가 수집되었다. 데이터는 자율신경계 반응과 주관적 감성을 측정하였나 TDP 를 이용한 분석과 적응적 TDP 분석방법으로 감성을 인식한 결과를 비교한 결과 평균 정확도는 증가하지 않았지만 전반적인 정확도 수준은 상승하는 것을 확인하였다. 그러므로 적응적 TDP를 이용할 경우 개인차를 줄일 수 있다는 것을 확인하였다.

• Journal of Applied Biological Chemistry
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• v.43 no.3
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• pp.136-140
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• 2000

A novel 6-deoxyhexose biosynthetic gene cluster different from the one for the biosynthesis of streptomycin was isolated from Streptomyces griseus using specifically designed PCR primers to compare the sequence of known dTDP-glucose synthase genes. We cloned a 5.8-kb DNA from Streptomyces griseus ATCC10137, which contained the 4-ketoreductase homologue (grsB), dTDP-glucose synthase (grsD), and dTDP-glucose 4, 6-dehydratase (grsE) genes. Escherichia coli cultures containing plasmid of the PCR product which encoded the grsE region under the controUed T7 promoter were able to catalyze the formation of dTDP-4-keto-6-deoxy-D-glucose from TDP-glucose. The enzyme showed high substrate specificity, being specific to only dTDP-glucose that is known to be incorporated into secondary metabolites such as antibiotics.

• Journal of Microbiology and Biotechnology
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• v.12 no.2
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• pp.217-221
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• 2002

dTDP-D-glucose 4,6-dehydratase (TDPDH) catalyzes the conversion of dTDP-D-glucose to dTDP-4-keto-6-deoxy-D-glucose, and requires $NAD^＋$ as a coenzyme for its catalytic activity. The dTDP-D-glucose 4,6-dehydratase from Streptomyces antibioticus $Tu{\ddot}99$ tightly binds $NAD^＋$ [19]. In order to determine the role of lysine-148 in the $NAD^＋$ binding, the lysine of the dTDP-D-glucose 4,6-dehydratase from Streptomyces antibioticus $Tu{\ddot}99$ was mutated to various amino acids by site-directed mutagenesis. The catalytic activity of the four mutated enzymes of TDPDH did not recover after addition of $NAD^＋$ . However, the activity of K159A, the mutated enzyme of UDP-D-glucose 4-epimerase (UDPE), recovered after the addition of $NAD^＋$ [15]. Although dTDP-glucose 4,6-dehydratase, and UDP-galactose (glucose) 4-epimerase are members of the short-chain dehydrogenase/reductase SDR family and the lysine-148 of TDPDH was highly conserved as in UDPE (Lys-159), the function of the lysine-148 of TDPDH was different from that of UDPE. The mutated enzymes showed that the lysine-148 of the dTDP-D-glucose 4,6-dehydratase played no role in the $NAD^＋$ binding. Accordingly, it is suggested that the lysine-148 of the dTDP-D-glucose 4,6-dehydratase is involved in the folding of TDPDH.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.576-583
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• 2004

GERI-155 is a macrolide antibiotic containing two deoxyhexose molecules and shows antimicrobial activities against Gram-positive bacteria. Deoxysugar biosynthetic gene cluster of GERI-155 from Streptomyces sp. GERI-l55 genome was cloned. Four orfs were identified and a putative orf presumed to be the dTDP g]ucose-4,6-dehydratase gene was designated as gerE. GerE was expressed in E. coli by using a recombinant expression vector pHJ1. The expressed protein was purified from E. coli cell lysate by using ammonium sulfate fractionation, and DEAE-sepharose CL-6B and hydroxylapatite column chromatography. The molecular mass of the expressed protein correlated with the predicted mass that was deduced from the cloned gene sequence data. The recombinant protein was a homodimer with a subunit relative molecular weight of 39,000 Dalton. It was found to have dTDP-glucose 4,6-dehydratase activity and also found to be highly specific for dTDP-glucose as a substrate. The values of $K_{m} and V_{max}$ for dTDP-g]ucose were $32\mu$M and 335 nmol $min^{-1}$ (mg protein)^{-1}$, respectively. dTTP and dTDP were strong inhibitors of the protein. $NAD^+$, the coenzyme for dTDP-glucose 4,6-dehydratase, was tightly bound to the expressed protein.

• Journal of Information Processing Systems
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• v.1 no.1 s.1
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• pp.70-74
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• 2005

Sensor networks have emerged as an interesting and important research area in the last few years. These networks require that time be synchronized more precisely than in traditional Internet applications. In this paper, we compared and analyzed the performance of the RBS and TDP mechanisms in the view of the number of generated messages and the synchronization accuracy. The reason that we chose be RBS ad the TDP mechanism to be compared is because the RES is an innovative method to achieve the high accurate synchronization. And TDP is a new method taking over the NTP method which has been used widely in the Internet. We simulated the performance of two methods assuming the IEEE 802.11 CSMA/CA MAC. As for the number of nodes in the sensor networks, two situations of 25 (for the small size network) and 100 (for the large size network) nodes are used. In the aspect of the number of messages generated for the synchronization, TDP is far better than RBS. But, the synchronization accuracy of RBS is far higher than that of TDP. We cm conclude that in a small size sensor networks requiring very high accuracy, such as an application of very high speed objects tracking in a confined space, the RBS is more proper than TDP even though the RBS may generate more traffic than TDP. But, in a wide range sensor networks with a large number of nodes, TDP is more realistic though the accuracy is somewhat worse than RBS because RBS may make so many synchronization messages, and then consume more energies at each node. So, two mechanisms may be used selectively according to the required environments, without saying that the one method is always better than the other.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.8
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• pp.1236-1242
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• 2007

Sixty healthy growing pigs ($Duroc{\times}Landrace{\times}Yorkshire$ with an average BW of 21.4 kg) were used to determine the true digestible phosphorus (TDP) requirement of growing pigs on the basis of growth performance and serum biochemical indices. Pigs were assigned randomly to one of five dietary treatments (12 pigs/diet), representing five levels of TDP (0.16%, 0.20%, 0.23%, 0.26% and 0.39%). There were three replications per treatment, with four pigs (2 barrows and 2 gilts) in each replication (2 pigs/pen) A randomized-block design was used, with pen as the experimental unit. Experimental diets were formulated to provide the 5 TDP levels with a total calcium (Ca) to TDP ratio of 2:1, and offered to pigs at 5% BW for 28 d. The total Ca contents of the five diets were 0.33, 0.38, 0.45, 0.51 and 0.79%, respectively. During the 28-d experimental period, the ADG of pigs was affected by dietary TDP levels as described by Equation 1: y = $-809,532x^4+788,079x^3-276,250x^2+42,114x-1$,759; ($R^2$ = 0.99; p<0.01; y = ADG, g/d; x = dietary TDP, %). The feed:gain ratio for pigs was affected by dietary TDP levels as described by Equation 2: y = $3,651.1x^4-3,480.4x^3+1,183.8x^2-172.5x+10.9$ ($R^2$ = 0.99; p<0.01; y = feed:gain ratio; x = dietary TDP, %). Total P concentrations in serum were affected by dietary TDP levels as described by Equation 3: y = $-3,311.7x^4+3,342.7x^3-1,224.6x^2+195.6x-8.7$ ($R^2$ = 0.99; p<0.01; y = total serum P concentration and x = dietary TDP, %). The highest ADG (782 g/d), the lowest feed:gain ratio (1.07), and the highest total serum P concentration (3.1 mmol/L) were obtained when dietary TDP level was 0.34%. Collectively, these results indicate that the optimal TDP requirement of growing pigs is 0.34% of the diet (e.g., 5.1 g/day for a 30-kg pig that consumed 1.5 kg feed daily) at a total Ca to TDP ratio of 2:1.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.83-86
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• 2002

Thermus caldophilus로부터 TDP-glucose 4,6-degydratase (TDPDH) 유전자를 분리하고 염기서열을 결정하고, 결정된 염기서열에 해당하는 유전자를 발현벡터에 삽입하고, 발현벡터를 대장균에 형질전환시켜 배양함으로써 TDP-glucose 4,6-dehydratase를 대량 생산하였다. 본 연구의 TDP-glucose 4,6-dehydratase는 주반응으로 TDP-4-keto-6-glucose를 합성하고, 여러 가지 당을 디옥시당으로 합성하는 효과가 있다.