• Journal of the Korean Wood Science and Technology
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• 제42권2호
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• pp.149-156
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• 2014

Termites are wood pests that cause vast economic damage every year. They digest both cellulose and starch, but the enzymes for starch digestion have not been well characterized. We obtained complete amino acid sequence information on the KME1 ${\alpha}$-amylase from Reticulitermes speratus KMT1 through analysis of total mRNA sequences. The KME1 enzyme has two ${\alpha}$-amylase domains and is 68% identical to the ${\alpha}$-amylase from Blattellager manica, its closest relative in the GenBank database. Some unique features of its conserved region and its distant evolutionary relationship to other insect ${\alpha}$-amylases suggest that KME1 is a new type of ${\alpha}$-amylase.

• 미생물학회지
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• 제22권3호
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• pp.167-173
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• 1984

The cellabiase (${\beta}$-glucosidase) gene in a Cellulomonas sp. CS1-1 was cloned into E. coli HB101 using the vector plasmid pBR322, and the expression of the gene in E. coli studied. The chromosomal DNA of the cellulomonas was digested by seveal restriction enzymes, each of which has only one cleaving site in plasmid pBR322. The recombinant plasmid, pSB2, created with Sal I frament, was expressed for the cellobiase gene in E. coli. The recombiant plasmid was estimated to contain 6.4 Kb foreign DNA at the Sal I site of plasmid pBR322 and the inserted DNA was mapped by single and double digestion with several enzymes. E. coli HB101(pSB2) has slowly grown in a mineral liquid medium containing cellobiose as a sole carbon source. The cellobiase activity in the transformed E. coli was 132 units per liter, which is equivalent to one twenty fifth of that in doner strain Cellulomonas sp. CS1-1. The transforned cell with plasmid containing cellulase gene grow well in the LB mediuns. The synthesis of cellobiase in the strain, E. coli HB101 (pSB2), was inhibited by glucose and at high concentration of cellobiose, and induced by cellobiose at low concentration.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.738-747
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• 2018

Objective: In a previous study, analysis of Illumina sequenced metagenomic DNA data of bacteria in Vietnamese goats' rumen showed a high diversity of putative lignocellulolytic genes. In this study, taxonomy speculation of microbial community and lignocellulolytic bacteria population in the rumen was conducted to elucidate a role of bacterial structure for effective degradation of plant materials. Methods: The metagenomic data had been subjected into Basic Local Alignment Search Tool (BLASTX) algorithm and the National Center for Biotechnology Information non-redundant sequence database. Here the BLASTX hits were further processed by the Metagenome Analyzer program to statistically analyze the abundance of taxa. Results: Microbial community in the rumen is defined by dominance of Bacteroidetes compared to Firmicutes. The ratio of Firmicutes versus Bacteroidetes was 0.36:1. An abundance of Synergistetes was uniquely identified in the goat microbiome may be formed by host genotype. With regard to bacterial lignocellulose degraders, the ratio of lignocellulolytic genes affiliated with Firmicutes compared to the genes linked to Bacteroidetes was 0.11:1, in which the genes encoding putative hemicellulases, carbohydrate esterases, polysaccharide lyases originated from Bacteroidetes were 14 to 20 times higher than from Firmicutes. Firmicutes seem to possess more cellulose hydrolysis capacity showing a Firmicutes/Bacteroidetes ratio of 0.35:1. Analysis of lignocellulolytic potential degraders shows that four species belonged to Bacteroidetes phylum, while two species belonged to Firmicutes phylum harbouring at least 12 different catalytic domains for all lignocellulose pretreatment, cellulose, as well as hemicellulose saccharification. Conclusion: Based on these findings, we speculate that increasing the members of Bacteroidetes to keep a low ratio of Firmicutes versus Bacteroidetes in goat rumen has resulted most likely in an increased lignocellulose digestion.

• Applied Biological Chemistry
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• 제36권2호
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• pp.105-110
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• 1993

Enterobacter sp. S45로부터 inulin fructotransferase를 생산, 정제하고 효소적 특성을 조사하였다. 배양상징액의 $0.4{\sim}0.8$ 포화$(NH_4)_2SO_4$ 침전물인 조효소는 DEAE-cellulose column chromatography 및 fast protein liquid chromatography로 정제하였으며 효소의 회수율은 0.9%였고 약 148배의 정제도를 보였다. 정제된 효소는 전기영동상으로 단일 band였으며 분자량은 SDS-PAGE에 의해 42,800으로 나타났다. 이 효소의 최적 pH는 5.5, 최적 온도는 $50^{\circ}C$였으며 $Mg^{2+}$이온은 효소활성을 30% 증가시키나 $Hg^{2+}$, $Cu^{2+}$ 및 $Fe^{3+}$이온들은 활성을 강력히 저해하였다. Inulin에 대한 km 값은 1.4 mM, Vmax 값은 $0.196\;{\mu}mole/min$이었다. 본 효소는 inulin을 fructose 말단으로부터 fructose 두 분자씩 절단, 환상형인 DFA를 생성하며, 그 결과 inulin 분해산물인 GF, $GF_2$, $GF_3$, $GF_4$ 등도 검출되었다. 중합도에 따른 효소활성을 조사해 본 결과 이 효소는 중합도 $4(GF_3)$ 이상의 fructo 올리고당에만 작용하여 $GF_3$는 DFA III와 GF로, $GF_4$는 DFA III와 $GF_2$로 변환시켰다. 또한 본 효소는 sucrose, raffinose 및 melezitose를 기질로서 이용하지 못하므로 invertase 및 ${\alpha}-glucosidase$ 활성이 없는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제21권8호
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• pp.808-817
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• 2011

Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.

• 미생물학회지
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• 제54권4호
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• pp.362-368
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• 2018

메뚜기는 광합성으로 고정된 탄소의 소화에서 중요한 역할을 한다. 장내 미생물 군의 도움으로, 메뚜기는 셀룰로오스 및 헤미셀룰로오스와 같은 잎의 성분을 분해할 수 있다. 본 연구는 한국의 기도에서 수집한 메뚜기 껍질에서 추출한 셀룰로오스 분해 효모 균주를 조사하기 위해 이루어졌다. 효모 균주 중 ON2와 ON17 (두 균주)과 ON6 (한 균주)는 CMC-플레이트 분석에서 셀룰로오스 활성을 보였다. Large subunit rDNA의 D1/D2영역의 서열과 internal transcribed spacer (ITS) 영역의 분석 결과, ON2와 ON17 균주가 Papiliotrema aspenensis CBS $13867^T$와 가장 밀접하게 관련되어 있었고(D1/D2 영역의 서열 유사성은 100% ITS에서 99.4%의 서열 유사성) ON6 균주는 Saitozyma flava와 관련된(D1/D2영역에서 100%, ITS에서 99.0%) 밀접하게 관련이 있었다. 이 세 가지 효모 균주는 모두 셀룰로오스를 분해할 수 있으므로 공생하는 효모들은 탄수화물 분해를 위한 효소를 자체적으로 생산하고 당 단당체를 휘발성 지방산으로 전환시킬 수 있다. Tremellomycetes에 속하는 공생 효모 균주인 ON2, ON17 (Papilioterma 속)과 ON6 (Saitozyma속)은 한국에는 보고되지 않은 균주이다.

• Asian-Australasian Journal of Animal Sciences
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• 제28권12호
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• pp.1669-1679
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• 2015

The quest to broaden the narrow range of feed ingredients available to pig producers has prompted research on the use of low cost, unconventional feedstuffs, which are typically fibrous and abundant. Maize cobs, a by-product of a major cereal grown worldwide, have potential to be used as a pig feed ingredient. Presently, maize cobs are either dumped or burnt for fuel. The major challenge in using maize cobs in pig diets is their lignocellulosic nature (45% to 55% cellulose, 25% to 35% hemicellulose, and 20% to 30% lignin) which is resistant to pigs' digestive enzymes. The high fiber in maize cobs (930 g neutral detergent fiber/kg dry matter [DM]; 573 g acid detergent fiber/kg DM) increases rate of passage and sequestration of nutrients in the fiber reducing their digestion. However, grinding, heating and fermentation can modify the structure of the fibrous components in the maize cobs and improve their utilization. Pigs can also extract up to 25% of energy maintenance requirements from fermentation products. In addition, dietary fiber improves pig intestinal health by promoting the growth of lactic acid bacteria, which suppress proliferation of pathogenic bacteria in the intestines. This paper reviews maize cob composition and the effect on digestibility of nutrients, intestinal microflora and growth performance and proposes the use of ensiling using exogenous enzymes to enhance utilization in diets of pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제12권7호
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• pp.1075-1079
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• 1999

Adult German cross $(German{\times}British{\times}Russian)$ angora rabbits (one year age), 32 in number were divided randomly into four groups $(T_1-T_4)$ with equal sex ratio and fed diets containing $T_1$ groundnut cake (GNC); $T_3$, soyaflakes (SF); $T_4$, sunflower cake (SFC) and $T_2$, a mixture of all the three cakes along with green forage as roughage for a period of 9 months. Nine per cent protein was added from each protein source. Fibre level was maintained by adjusting the level of rice phak in the diets. The diets were iso-nitrogenous and contained similar level of fibre. DMI through roughage was not affected due to source of protein in the diet, however, DMI through concentrate was higher $(p{\leq}0.05)$ with SFC diet, which resulted in higher total feed intake in the group $(T_4)$. Body weights increased up to second shearing, thereafter it decreased due to summer depression. Diet containing soyaflakes sustained higher wool yield whereas, it was lowest $(p{\leq}0.05)$ on SFC diet. Wool attributes (staple length, medullation, fibre diameter) were not affected due to source of protein in the diet. Digestibility of fibre and its fractions (ADF, cellulose, hemicellulose) decreased $(p{\leq}0.05)$ with incorporation of SFC in the diets. Balance of calcium was lowest whereas, that of nitrogen was highest with SFC diet $(T_4)$. Biological value of N and net protein utilization was better when different protein sources were mixed together $(T_2)$. Protein quality of soyaflakes proved better for wool production followed by groundnut cake and mixture of three protein sources. Sunflower cake alone or in combination decreased wool production which may be checked by supplementation of amino acids and energy.

• BMB Reports
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• 제44권1호
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• pp.52-57
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• 2011

Termites play an important role in the degradation of dead plant materials and have acquired endogenous and symbiotic cellulose digestion capabilities. The feruloyl esterase enzyme (FAE) gene amplified from the metagenomic DNA of Coptotermes formosanus gut was cloned in the TA cloning vector and subcloned into a pET32a expression vector. The Ft3-7 gene has 84% sequence identity with Clostridium saccharolyticum and shows amino acid sequence identity with predicted xylanase/chitin deacetylase and endo-1,4-beta-xylanase. The sequence analysis reveals that probably Ft3-7 could be a new gene and that its molecular mass was 18.5 kDa. The activity of the recombinant enzyme (Ft3-7) produced in Escherichia coli (E.coli) was 21.4 U with substrate ethyl ferulate and its specific activity was 24.6 U/mg protein. The optimum pH and temperature for enzyme activity were 7.0 and $37^{\circ}C$, respectively. The substrate utilization preferences and sequence similarity of the Ft3-7 place it in the type-D sub-class of FAE.

• 대한간호학회지
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• 제25권3호
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• pp.457-471
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• 1995

This research was done to promote improvement of practical application of nursing diagnoses and to improve the quality of nursing. The subjects of this research were 156 second year students of C junior nursing college who were giving adult patient care. The nursing diagnoses of 312 reports were analyzed using NANDA. In these case reports only nursing diagnoses were considered, of which there were a total of 982. In the data analysis the 9H of the nursing students' nursing diagnoses matched with 105 NANDA nursing diagnoses, Of these, the most frequent diagnoses were pain(165, 17.48%), anxiety(101, 10.70%), alteration in nutrition(83, 8.79%) , sleep disturbance (67, 7.10%), in activity intolerance (67, 7.10%), ineffective breathing pattern(51,5.40%). The etiology for the students' nursing diagnoses were compared with NANDA's nursing diagnoses by frequency. The most frequent etiology for the nursing diagnoses of pain was a biological etiology(50, 31%), for anxiety, situation crisis(58, 57.43%), for alteration in nutrition, indigesion(23, 27.71%), for sleep disturbance, external etiology(25, 37.32%), for activity intolerance, immobile position(22, 32.84%), for ineffective breathing pattern, pain(35, 68.63%), and for ,impaired physical mobility, pain(31, 65.96%). The most frequent etiology for constipation was inadquate digestion of water and cellulose (16, 34.78%), for fluid volume felicity, loss of body fluid (21, 52.50%), for impaired skin integrity, external etilogy(16, 43.24%), for impaired physical mobility, pain(22, 62.86%) , for knowledge deficits, cognition disturbance(9, 27.27％), for ineffective air way clearance, secretion obstruction(14, 48.27%) , for impaired gas exchange, loss of transport ability of blood oxygen(9, 37.50%) , and for powerlessness, therapy environment (5, 22.73%). The number of nursing diagnoses by pattern was exchange(16), moving(6), feeling(4), choosing(4), relating(3), communication(1), perceiving(1), knowing(1), valuing(1).