Substrate specificity of a recombinant d-lyxose isomerase from Providencia stuartii for monosaccharides

Kwon, H.J.,Yeom, S.J.,Park, C.S.,Oh, D.K. Society for Bioscience and Bioengineering, Japan ; 2010 Journal of bioscience and bioengineering Vol.110 No.1

The specific activity and catalytic efficiency (k<SUB>cat</SUB>/K<SUB>m</SUB>) of the recombinant putative protein from Providencia stuartii was the highest for d-lyxose among the aldose substrates, indicating that it is a d-lyxose isomerase. Gel filtration analysis suggested that the native enzyme is a dimer with a molecular mass of 44 kDa. The maximal activity for d-lyxose isomerization was observed at pH 7.5 and 45 <SUP>o</SUP>C in the presence of 1 mM Mn<SUP>2+</SUP>. The enzyme exhibited high isomerization activity for aldose substrates with the C2 and C3 hydroxyl groups in the left-hand configuration, such as d-lyxose, d-mannose, l-ribose, d-talose, and l-allose (listed in decreasing order of activity). The enzyme exhibited the highest activity for d-xylulose among all pentoses and hexoses. Thus, d-lyxose was produced at 288 g/l from 500 g/l d-xylulose by d-lyxose isomerase at pH 7.5 and 45 <SUP>o</SUP>C for 2 h, with a conversion yield of 58 % and a volumetric productivity of 144 g l<SUP>-1</SUP> h<SUP>-1</SUP>. The observed k<SUB>cat</SUB>/K<SUB>m</SUB> (920 mM<SUP>-1</SUP> s<SUP>-1</SUP>) of P. stuartiid-lyxose isomerase for d-xylulose is higher than any of the k<SUB>cat</SUB>/K<SUB>m</SUB> values previously reported for sugar and sugar phosphate isomerases with monosaccharide substrates. These results suggest that the enzyme will be useful as an industrial producer of d-lyxose.

Substrate specificity of a recombinant <small>D</small>-lyxose isomerase from <i>Serratia proteamaculans</i> that produces <small>D</small>-lyxose and <small>D</small>-mannose

Park, C.-S.,Yeom, S.-J.,Lim, Y.-R.,Kim, Y.-S.,Oh, D.-K. Blackwell Publishing Ltd 2010 Letters in applied microbiology Vol.51 No.3

<P>Abstract</P><P>Aims: </P><P>Characterization of substrate specificity of a <SMALL>D</SMALL>-lyxose isomerase from <I>Serratia proteamaculans</I> and application of the enzyme in the production of <SMALL>D</SMALL>-lyxose and <SMALL>D</SMALL>-mannose.</P><P>Methods and Results: </P><P>The concentrations of monosaccharides were determined using a Bio-LC system. The activity of the recombinant protein from <I>Ser. proteamaculans</I> was the highest for <SMALL>D</SMALL>-lyxose among aldoses, indicating that it is a <SMALL>D-</SMALL>lyxose isomerase. The native recombinant enzyme existed as a 54-kDa dimer, and the maximal activity for <SMALL>D-</SMALL>lyxose isomerization was observed at pH 7·5 and 40°C in the presence of 1 mmol l<SUP>−1</SUP> Mn<SUP>2+</SUP>. The <I>K</I><SUB>m</SUB> values for <SMALL>D</SMALL>-lyxose, <SMALL>D</SMALL>-mannose, <SMALL>D</SMALL>-xylulose, and <SMALL>D</SMALL>-fructose were 13·3, 32·2, 3·83, and 19·4 mmol l<SUP>−1</SUP>, respectively. In 2 ml of reaction volume at pH 7·5 and 35°C, <SMALL>D</SMALL>-lyxose was produced at 35% (w/v) from 50% (w/v) <SMALL>D</SMALL>-xylulose by the <SMALL>D-</SMALL>lyxose isomerase in 3 h, while <SMALL>D</SMALL>-mannose were produced at 10% (w/v) from 50% (w/v) <SMALL>D</SMALL>-fructose in 5 h.</P><P>Conclusions: </P><P>We identified the putative sugar isomerase from <I>Ser. proteamaculans</I> as a <SMALL>D</SMALL>-lyxose isomerase. The enzyme exhibited isomerization activity for aldose substrates with the C2 and C3 hydroxyl groups in the left-hand configuration. High production rates of <SMALL>D-</SMALL>lyxose and <SMALL>D</SMALL>-mannose by the enzyme were obtained.</P><P>Significance and Impact of the Study: </P><P>A new <SMALL>D-</SMALL>lyxose isomerase was found, and this enzyme had higher activity for <SMALL>D</SMALL>-lyxose and <SMALL>D</SMALL>-mannose than previously reported enzymes. Thus, the enzyme can be applied in industrial production of <SMALL>D</SMALL>-lyxose and <SMALL>D</SMALL>-mannose.</P>

Production of d-psicose from d-fructose by whole recombinant cells with high-level expression of d-psicose 3-epimerase from Agrobacterium tumefaciens

Park, C.S.,Park, C.S.,Shin, K.C.,Oh, D.K. Society for Bioscience and Bioengineering, Japan ; 2016 Journal of bioscience and bioengineering Vol.121 No.2

<P>The specific activity of recombinant Escherichia coli cells expressing the double-site variant (133L-S213C) D-psicose 3-epimerase (DPEase) from Agrobacterium tumefaciens was highest at 24 h of cultivation time in Terrific Broth (TB) medium among the media tested. The contents of crude protein and DPEase in recombinant cells at 24 h were 37.0 and 8.6% (w/w), respectively, indicating that the enzyme was highly expressed. The reaction conditions for the production of D-psicose from D-fructose by whole recombinant cells with the highest specific activity were optimal at 60 degrees C, pH 8.5, 4 g/l cells, and 700 g/l D-fructose. Under these conditions, whole recombinant cells produced 230 g/I D-psicose after 40 min, with a conversion yield of 33% (w/w), a volumetric productivity of 345 g/l/h, and a specific productivity of 86.2 g/g/h. These are the highest conversion yield and volumetric and specific productivities of D-psicose from D-fructose by cells reported thus far. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.</P>

Influence of supplemental canola or soybean oil on milk yield, fatty acid profile and postpartum weight changes in grazing dairy goats

Israel Lerma-Reyes,German D. Mendoza-Martinez,Rolado Rojo-Rubio,Mario Mejia,J.C. Garcia-Lopez,Hector A. Lee-Rangel 아세아·태평양축산학회 2018 Animal Bioscience Vol.31 No.2

Objective: This experiment was designed to evaluate the effect of supplementation with soybean or canola oil on milk production and the composition of long chain fatty acids as well as weight changes in the goats and their kids. Methods: Thirty nine mulitparous crossed Alpine×Nubian goats (initial body weight [BW] 43.5±1.7 kg) from the day of parturition were assigned to the treatments: grazing control (n = 15); grazing plus 20 mL/goat/d of supplemental soybean oil (n = 12); and grazing plus 20 mL/goat/d of supplemental canola oil (n = 12) from November 26, 2014 to March 7, 2015. The planned contrasts were: CI (control vs supplemented with oils); CII (soybean vs canola oil) to compare the treatment effects. Results: The vegetable oil supplementation reduced weight losses in lactating goats (CI: –0.060 vs 0.090 kg/d; p = 0.03) but did not improve milk production or affect kids’ growth. The content of C4, C6, C8, C10, C11, C14, and C18:1n9t in the milk was increased (p<0.05) with respect to control. However, C12, C14, C16, C18, C18:1n9c, C18:2n6c, and C18:3n3 were reduced (p<0.05) in supplemented goats. Conjugated linoleic acid (CLA) was increased (p<0.05) in goats supplemented with oils compared to the control group. Conclusion: Supplementation with 20 mL/d of soybean or canola oil did not affect milk production or kids’ performance; however, it increased CLA concentration and reduced the reduced weight losses in lactating goats.

C3, C4 의 Nephelometric 측정 경험

조병철 ( B C Cho ),검덕희 ( D H Kim ),김민숙 ( M S Kim ),서덕규 ( D K Seo ) 대한임상검사과학회 1990 대한임상검사과학회지(KJCLS) Vol.22 No.1

RIA has been used to measure accurately a minute quantity of Ags, such as proteins. hormones, enzymes or drugs. However, due to the presence of a radication hazard and the increment of having to pool test specimens. RIA is gradually being replaced by other immunoassay that have a comparable sensitivity but do not require radiocative meterial. Turbidimetric immunoassay, latex immunoassay and fluorescence immunoassay are such examples. We at the department of clinical pathology, Asan medical center, compared CH 50 activity with C3 , C4 quantity by nephelometric method.

소 c-KIT Receptor 유전자의 다형성에 관한 연구

장요순,김태헌,윤두학,박응우,이혜원,이학교,정일정 한국동물자원과학회 2002 한국축산학회지 Vol.44 No.6

소의 흰 반점 관련 후보유전자로 c-KIT receptor 유전자를 선정하여, c-KIT receptor 유전자내의 변이를 탐색하고 변이가 흰반점 표현형과 연관성이 있는지를 분석하였다. 한우, Angus, Brown Swiss, Charolais, Hereford, Holstein, Limousin 및 Simmental 등 8개 품종의 DNA 시료를 사용하여 c-KIT receptor 유전자의 intron 6번 영역에서 다형성을 조사하고 분석하였다. c-KIT receptor 유전자의 intron 6번 영역에서는 4개의 염기치환이 발견되어, MspⅠ, BsrBⅠ 및 NdeⅠ 제한효소를 이용하여 PCR-RFLP 분석을 실시하였다. Intron 6번을 포함하는 영역의 PCR 산물 크기는 2,440 bp 이었다. MspⅠ다형성은 PCR-RFLP 분석 결과 3개의 대립유전자가 존재하였으며, 한우품종에서는 3개의 대립유전자 모두가 발견되었고, CC 형태이 유전자형을 제외한 5개의 유전자형 (AA, AB, AC, BC 및 BB)을 확인하였다. Angus, Brown Swiss, Hereford, Holstein 및 Simmental 품종에서는 A 대립유전자만을 갖는 것으로 조사되었고, 한우는 44%만 AA 유전자형을 나타내었다. BsrBⅠ 다형성은 2개의 대립유전자로서 3개의 유전자형이 나타나는 것을 확인하였으며, Charolais 및 Hereford 품종이 다른 소 품종에 비하여 A 대립유전자의 빈도가 높게 나타났다. NdeⅠ 다형성을 분석한 결과 Brown Swiss 품종에서는 NdeⅠ에 의해 절단되는 형태인 A 대립유전자만 관찰되었으며, Holstein 품종은 92%, Simmental 품종은 72%가 절단되는 형태를 나타내어, 모색이 흰색을 띠는 소 품종에서 절단되는 형태가 많았다. 소 c-KIT receptor 유전자의 intron 6번 영역에서 확인된 4개의 염기치환은 품종에 따라 다른 빈도를 보였으나, 이들 염기치환과 흰 반점과의 연관성에 대한 증거는 발견하지 못하였다. 그러므로 소의 흰 반점과 c-KIT receptor 유전자 내의 변이와의 관련성은 다른 영역에 대한 추가적인 분석과, 이미 보고된 다른 모색관련 유전자의 다형성과의 연관성 분석 등과 같은 연구가 필요한 것으로 판단된다. We considered KIT gene as a candidate gene for the white-spotting pattern in cattle. This study was carried out to detect genetic variation of c-KIT receptor gene and to investigate association between the mutation and the white-spotting pattern in cattle. PCR-RFLP analysis within intron 6 of c-KIT receptor gene were performed with 8 cattle breeds including Hanwoo, Angus, Brown Swiss, Charolais, Hereford, Holstein, Limousin and Simmental. When PCR product of approximately 2,440 bp including intron 6 of c-KIT receptor gene was sequenced, four nucleotide substitutions were found within intron 6 of the bovine c-KIT receptor gene. In PCR-RFLP analysis, three alleles (A, B and C), two alleles (A and B) and two alleles (A and B) at each locus were identified by Msp Ⅰ, BsrBⅠ and NdeⅠ, respectively. Although frequencies of allele at each locus were different among cattle breeds, we could not get any evidence related with white or white spotting phenotypes in these mutations on intron 6 of c-KIT receptor gene. However, we can not entirely exclude the possibility that c-KIT receptor gene is responsible for white spotting phenotype in cattle. Thus, further studies need to detect other mutations in c-KIT receptor gene and to test association of those mutations and coat color phenotypes in cattle.

주성분 분석을 이용한 거세한우의 체형분류에 관한 연구

하동우,김현철,김병우,이문연,이종헌,신철교,도창희,이정규 한국동물자원과학회 2002 한국축산학회지 Vol.44 No.6

1997년부터 2001년까지 (사)한국종축개량협회의 전국한우능력평가대회에 출품된 642두의 거세한우의 도축 전까지 4회에 걸쳐 주요 10개 부위의 체형 측정치를 조사하여 주성분 분석을 이용하여 체형을 분류하였으며, 각 체형 측정치의 주성분 지수와 도체 형질과의 상관관계를 분석하였으며, 그 결과를 요약하면 다음과 같다. 1. 1차, 2차, 3차 및 4차 체형 측정치의 principal component(이하 P.C) 1의 분산은 각각 7.610, 8.297, 7.269 및 5.736로서 전체분산 중 76.0%, 83.0%, 72.7% 및 57.4%를 나타낼 수 있었고, P.C 1, P.C 2, P.C 3의 합은 전체분산 중 각각 86.69%, 90.49%, 84.62% 및 77.26%를 나타났다. 2. 각 차수별 체형 측정치의 P.C 1의 모든 계수가 양(+)이어서 일반적으로 좋은 체구의 크기를 나타내었으며, 체구의 크기에 가장 크게 영향을 미치는 형질은 흉심(0.328∼0.339)과 요각폭(0.325∼0.341)이였다. 또한 P.C 2, P.C 3 및 P.C 4는 각 차수별로 다양하게 나타났다. 3. 1차, 2차, 3차 및 4차 체형 측정치의 주성분 지수들과 도체형질간의 상관계수 추정에서 대부분의 분산을 차지하는 P.C 1은 도체중(0.539∼0.755), 일당증체량(0.256∼0.564), 등지방두께(0.227∼0.280) 및 배최장근단면적(0.187∼0.344)간에 정(+)의 상관을 보였으며, 육량지수(-0.246∼-0.110)와는 부(-)의 상관을 보였고, 근내지방도(0.066∼0.099)와는 대부분 유의적인 상관을 보이지 않거나 낮은 상관을 보였고, P.C 2, P.C 3 및 P.C 4는 주성분 지수들의 값에 따라 도체형질과 다양한 상관관계를 보였다. 단일 체형형질과 도체형질간의 상관 그리고 P.C 1의 주성분 지수와 도체형질과의 상관간에는 큰 차이가 없었다. Data were consisted of the ten body measurements (withers height, rump height, body length, chest depth, chest width, rump width, rump length, thurls width, hipbone width and chest girth) of 642 steers (Korean cattle), which was entered in the National Beef Quality Contest hosted by the Korea Animal Improvement Association from 1997 to 2001. A principal components analysis was used to classify the body types of the steers, and estimate the correlations between carcass traits and principal components for the body measurements of the first, second, third and fourth period, respectively. The first principal component of body measurements at the first, second, third and fourth period accounted for 76.0%, 83.0%, 72.7% and 57.4% of the total variance, respectively. The sum of first, second and third principal component at each period accounted for 86.69%, 90.49%, 84.62% and 77.26% of the total variance, respectively. At each period, all the first principal component of the body measurements were positive and it generally showed large framed body shape. The size of body was influenced mostly by chest depth(0.328∼0.339) and rump length(0.325∼0.341). The second, third and fourth principal component at the each period were various. There were positive correlations between principal components index of each period and carcass traits such as carcass weight(0.539∼0.755), average daily gain(0.256∼0.564), backfat thickness(0.227∼0.280), and eye muscle area(0.187∼0.344). The correlation with yield grade index(-0.246∼-0.110), however, was negative. The correlation with marbling score(0.066∼0.099) was low or statistically insignificant. According to principal component indexes of the second, third, and fourth components, the correlations with the carcass traits were various. There were no large differences between the correlations of the single body measurement trait with the carcass traits and the correlations of the first principal component indexes with the carcass traits.

Effect of Feeding Cyanidin 3-glucoside (C3G) High Black Rice Bran on Nutrient Digestibility, Blood Measurements, Growth Performance and Pork Quality of Pigs

Kil, D.Y.,Ryu, S.N.,Piao, L.G.,Kong, C.S.,Han, S.J.,Kim, Y.Y Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.12

Two experiments were conducted to investigate the effect of feeding cyanidin 3-glucoside (C3G) high black rice bran on nutrient digestibility, blood measurements, growth performance and pork quality of pigs. In Exp. I, a total of fifteen pigs (19.91${\pm}$1.80 kg, average initial body weight) were used in assay of nutrient digestibility and blood measurements. All pigs were allotted to 5 treatments with 3 replicates according to a completely randomized design (CRD) in an individual metabolic crate. Treatments included 1) CON: basal diet, 2) BRB-2: basal+brown rice bran 2%, 3) BRB-4: basal+brown rice bran 4%, 4) CRB-2: basal+C3G high black rice bran 2% and 5) CRB-4: basal+C3G high black rice bran 4%. The digestibility of dry matter (DM), crude protein (CP), crude fat (CF), crude ash (CA) and crude fiber (CF) was not affected by dietary treatments. Serum triglyceride (TG) and high density lipoprotein (HDL) cholesterol concentrations were not affected by addition of C3G high black rice bran. However, at the end of experiment, pigs fed rice bran showed decreased tendency in total cholesterol concentration. Especially pigs fed C3G high black rice bran showed significantly lower total cholesterol concentration compared to pigs fed brown rice bran (p<0.03). There was numerically lower total cholesterol concentration with increasing levels of black rice bran in the diet. In terms of serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT), there were no significant differences among treatments, even though pigs fed CRB-4 showed the lowest GOT concentration compared to other pigs. In Exp. II, sixteen finishing pigs (average initial body weight 89.96${\pm}$0.35 kg) were divided into 4 treatments to investigate the effect of feeding C3G high black rice bran on growth performance and pork quality. There were no significant differences in average daily gain (ADG), average daily feed intake (ADFI) and feed conversion ratio (FCR) among the treatments. Pigs fed C3G high black rice bran showed numerical decrease in ADG and increase in FCR while not effecting feed intake. There was no significant difference in live weight, carcass weight, carcass rate, backfat thickness and carcass grade. However, pigs fed C3G high black rice bran tended to show lower backfat thickness than pigs fed basal diet. Pigs fed C3G high black rice bran showed a tendency of decreased TBA value than pigs fed basal diet, although there was no overall significant difference among treatments. In conclusion, nutrient digestibility, blood measurements, growth performance and pork quality were not significantly affected by feeding C3G high black rice bran to pigs. However, C3G high black rice bran might have an effect on lowering serum total cholesterol and decrease the TBA value in pork compared to control group and these effects might be due to high concentration of antioxidative compounds in C3G high black rice bran.

Shewanella marina sp. nov., isolated from seawater

Park, S. C.,Baik, K. S.,Kim, M. S.,Kim, D.,Seong, C. N. Microbiology Society 2009 International journal of systematic and evolutiona Vol.59 No.8

<P>A motile, rod-shaped, pale-brown-pigmented bacterium, designated strain C4T, was isolated from seawater collected from the South Sea (Republic of Korea). Cells were Gram-negative, facultatively anaerobic, and catalase- and oxidase-positive. The major fatty acids were summed feature 3 (C16:1omega7c and/or iso-C15:0 2-OH; 19.4%), C16:0 (16.3%), C17:1omega8c (9.5%) and iso-C15:0 (7.7%). The DNA G+C content was 40.8 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain C4T formed a lineage within the genus Shewanella (92.7-96.1% sequence similarity to representative strains of the genus Shewanella) and was part of a distinct branch with the clade comprising Shewanella haliotis DW01T and Shewanella algae ATCC 51192T. Phenotypic characteristics enabled strain C4T to be distinguished from S. haliotis and S. algae. On the basis of the data presented in this study, strain C4T represents a novel species, for which the name Shewanella marina sp. nov. is proposed. The type strain is C4T (=KCTC 22185T=JCM 15074T).</P>

Development of additive [¹¹C]CO₂ target system in the KOTRON-13 cyclotron and its application for [¹¹C]radiopharmaceutical production

Moon, B.S.,Lee, H.J.,Lee, W.K.,Hur, M.G.,Yang, S.D.,Lee, B.C.,Kim, S.E. North-Holland Physics Pub 2015 Nuclear instruments & methods in physics research. Vol.356 No.-

The KOTRON-13 cyclotron, which was developed in South Korea for the production of medical radioisotopes, has the structural limitation of only one beam-output port, restricting the production of the carbon-11 isotope. In the present study, we investigate the design of a switchable target system and develop an effective carbon-11 target in the KOTRON-13 cyclotron, for combination with the fluorine-18 target. The target system was designed by introducing a sliding-type element between the fluorine-18 and carbon-11 targets, a tailor-made C-11 target and its cooling system. For the efficient production of [<SUP>11</SUP>C]CO<SUB>2</SUB>, the desirable target shape and internal volume were determined by a Stopping and Range of Ions in Matter (SRIM) simulation program, and the target grid was modified to resist the cavity pressure during beam irradiation. We evaluated the [<SUP>11</SUP>C]CO<SUB>2</SUB> production while varying the material and thickness of the target foil, oxygen content of the nitrogen gas, and target loading pressure. Using sliding-type equipment including an additional gate valve and a high vacuum in a beam line, the bi-directional conversion between the fluorine-18 and carbon-11 targets was efficient regarding the accurate beam irradiation on both targets. The optimal [<SUP>11</SUP>C]CO<SUB>2</SUB> production for 30min irradiation at 60μA (86.6+/-1.7GBq in the target at EOB) was observed at a thickness of 19μm with HAVAR® material as a target foil and a target loading pressure of 24bar with nitrogen plus 300ppb of oxygen gas. Additionally, the coolant cavity system in the target grid and target chamber is useful to remove the heat transferred to the target body by the internal convection of water and thereby ensure the stability of the [<SUP>11</SUP>C]CO<SUB>2</SUB> production under a high beam current. In the application of C-11 labeled radiopharmaceuticals such as [<SUP>11</SUP>C]PIB, [<SUP>11</SUP>C]DASB, [<SUP>11</SUP>C]PBR28, [<SUP>11</SUP>C]Methionine and [<SUP>11</SUP>C]Clozapine, the radiochemical yields were shown to be 25-38% (decay corrected) with over 166GBq/μmol of specific activity. Consequently, the additive carbon-11 target system was successfully developed in only one output port of the KOTRON-13 cyclotron and exhibited the stable production of C-11 labeled radiopharmaceuticals.