LMW-GSs represent approximately 1/3 of the total wheat gluten fraction, which have not been widely studied, even though they are important in the context of wheat end-use quality. In this study, we report on the qualitative and quantitative analysis of LMW-GS in korean wheat cultivars by 2DE in 32 cultivars of Korean wheat for the use of the basis of wheat breeding. We firstly identified spots corresponding each of Glu-3 alleles. The 2DE results for each cultivar will be used as reference map or protein marker discriminating wheat cultivars, wheat and rice, imported and Korean flour. Unexpectedly, five LMW-GS spots were found to be expressed at a common position in hexaploid wheat cultivars, and these spots might play something in glutenin biosynthesis. Total spot numbers were expressed variously between 20 and 10, and average spot number was shown 17.12. The average number of spots in Glu-A3, Glu-B3 and Glu-D3 were 3.0, 4.56 and 2.96 respectively. When the Korean wheat cultivars were compared with the Chinese spring (1.0) in the average relative expression level, Korean one’s were lower as 0.67. Especially, Gobun was the highest as 1.32 and Baekjoong was the lowest as 0.24. Also we investigated phylogenetic relationship based on frequency of HMW-GS spots and quantification value of each spot to all LMW-GS spots. As a result, the varieties of Korean wheat could be classified into five groups.

To evaluate expression level of HMW-GS protein qualitatively and quantitatively, we separated glutenin fractions and conducted two-dimensional electrophoresis (2DE) in 32 cultivars of Korean wheat for the use of as the basis of wheat breeding. The average spot number of HMW-GS in all Korean wheat cultivars was 11.78 which included 1.31, 5.53 and 4.94 to Glu-A1, Glu-B1 and Glu-D1 loci, respectively. Cultivars harboring 1, 2* subunits had many spots more than ones harboring null allele in Glu-A1 loci because there is no difference of spots between Glu-B1 and Glu-D1 loci. In total spot number of HMW-GS, the highest one was Jokyung as 18 and Dahong the lowest as 7. When the Korean wheat cultivars were compared with the Chinese spring in the average relative expression level, Korean one’s were lower as 0.44. Especially, Gobun was the highest as 1.11 and Eunpa was the lowest as 0.24. Also we investigated phylogenetic relationship based on both frequency of HMW-GS spots and quantification value of each spot to all HMW-GS spots. As a result, Korean the varieties of Korean wheat could be classified into six groups.

국내 밀 품종 조경, 금강 그리고 중국 밀 품종인 Chinese spring의 genomic DNA를 주형으로 LMW-GS 특이 프라이머세트를 이용하여 3개의 새로운 LMW-GS i 타입 유전자를분리하였고 이들의 분리된 유전자는 각 각 조경 II-2, CSIII-5 그리고 금강 6-12로 명명하였다. 이들의 유추 아미노산을 분석한 결과 20개의 시그널 펩타이드, 이소루신으로 시작하는 N-말단 부분 그리고 글루타민이 많은 반복도메인 그리고 C-말단 부분으로 구성되어 있으며 조경 II-2와 CS III-5는 전형적인 LMW-GS i-type 유전자처럼 C-말단에 8개의 시스테인 잔기가 있었다. 금강 6-12는 특이하게도 하나 더 많은 9개의 시스테인 잔기가 존재하였는데 이 여분의 시스테인 잔기는7번째 시스테인 잔기의 11잔기 앞에 존재하며 TAT(타이로신)이 TGT(시스테인)로 바뀐 결과이다. LMW-i 타입 글루테닌 유전자들 간의 SNP와 InDel을 확인하기 위해서 본 연구에서 클로닝 된 조경 II-2, CS III-5 그리고 이전에 본 그룹에서 확인된 조경 HQ619933와 기존 문헌에 나와 있는 6배 체 밀 유래의 10개의 LMW-GS i 타입 유전자들과 다중염기서열 분석을 실시하였고, 이들 사이에서 15개의 SNP와 1개의 insertion이 확인되었다. 밀 품종 조경의 Glu-A3 단백질을 동정하기 위해 글루테닌을 추출 이차원전기영동을 하고 Glu-A3c 위치의 스팟을 절취하여 in-gel digestion한 후 LC-ESI MS/MS 분석을 수행한 결과 조경의 i 타입 LMW-GS 유전자 좌는 Glu-A3c로 확인되었다. LMW-i 타입 글루테닌 유전자들의 연관 관계를 분석하기 위해 본 연구 그룹에서 클로닝 한 조경 II-2, CS III-5, 금강 6-12 그리고 조경 HQ6199333와 Genebank DB의 35개의 LMW-i 타입 글루테닌 유전자의 유추 아미노산 서열을 이용하여 Phylogenic tree를 완성하였다. 이들 39개의 계통도 분석 결과 이배체 밀과 4배체 밀의 LMi 타입 글루테닌이 육배체 밀의 LMW-i 타입 글루테닌과 크게 나눠지는 것을 확인하였으며, 육배체 밀의 LMW-i 타입 글루테닌들은 Glu-A3a부터 GluA-3g까지 7개 subgroup으로 나눠지는 것을 확인하였다. 금강 6-12는 GluA-3a와 GluA-3c 사이에 존재하였고 조경 II-2와 CS III-5는 GluA-3d와 일본 연질 밀인 농림 61의 AB062878과 같은 subgroup에 존재하였고 조경 HQ6199333은 Glu-A3c subgroup에 위치하였다. LMW-i 타입 글루테닌 유전자들의 유추 아미노산 다중서열분석결과 반복 도메인은 length polymorphism은 179～149개 정도의 long 타입과 91, 51, 10, 2개의 short 타입으로 나눠지고 이것은 long 타입과 short 타입 LMW-i 타입 글루테닌 유전자를 구분 할 수 있는 마커의 근거가 된다.

최근에는 콜라겐 펩타이드가 항상화효과를 통해 피부의 재생을 촉진시키는 효과를 한다는 연구가 보고되고있다. 본 구는 저분자 경구 콜라겐 펩타이드가 1550 nm fractional photothermolysis 레이저 치료 후 피부재생에 미치는 영향을 알아보고자 한다. 본 실험에 참여한 환자는 총 10명으로 실험군과 대조군 각 5명씩 무작위배정되었다. 대조군과 실험군 모두 동일하게 fractional photothermolysis 레이저를 시술받았다. 실험군에서는 경구 콜라겐 펩타이드를 하루에 1,000 mg씩 총 8주간 복용하였다. 치료 전후 모든 환자에서 피부의 탄성도, 경표피수분손실도, 홍반지수를 측정하였다. 피부탄성도는 실험군에서 대조군에 비해 통계적으로 유의하게 피부의 탄력성이 증가하였음을 확인하였다(p < 0.05). 경표피수분손실도는 두군간에 유의한 차이가 없었고, 홍반지수는 실험군에서 더 빨리 감소하는 결과를 보였다. 환자의 주관적 만족도평가에서 시술 4주 후 환자의 만족도는 실험군에서 평균 2.0점, 대조군에서 1.2점으로 나타났다. 저분자 콜라겐 펩타이드는 1550 nm fractional photothermolysis 레이저 치료 후 피부회복 촉진에 도움이 된다.

In order to improve rice dough functionality, we cloned 4 kinds of high-molecular-weight glutenin subunit (HMW-GS) genes from bread wheat, ‘Jokyeong’. Among them, we first examined Dx5 gene to generate marker-free transgenic rice for advanced quality processing of bread and noodles. The GluB1 promoter was inserted into binary vector for seed specific expression of the Dx5 gene. Two expression cassettes comprised of separate DNA fragments containing only the high-molecular-weight glutein subunit (HMW-GS) protein (Dx5) and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Dx5 or HPTII was infected to rice calli at 3: 1 ratio of Dx5 and HPTII, respectively. Then among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted both with Dx5 and HPTII gene to rice genome. We reconfirmed integration of the Dx5 and HPTII genes into the rice genome by Southern blot analysis. Wheat Dx5 transcriptsin rice seeds was examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only Dx5 gene were successfully screened at T1 generation. This result also provides that co-infection system with two expression cassettes could be efficient strategy to generate marker-free transgenic rice plants.

Gluten is the main functional component of wheat, and is the main source of the viscoelastic properties in a dough. One of the gluten group is glutenin, which is composed of high molecular weight (HMW) and low molecular weight (LMW) subunits. The HMW glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the grain. They are encoded by the Glu-1 loci located on the long arms of homeologous group one chromosomes, with each locus comprising two genes encoding x- and y-type subunits. The presence of certain HMW subunits is positively correlated with good bread-making quality. The highly conserved N- and C- terminal contaning cystein residues which form interand intra-chain disulphide bonds. This inter chain disulphide bonds stabilize the glutenin polymers. In contrast, the repetitive domains that comprise the central part of the HMW-GS are responsible for the elastic properties due to extensive arrays of interchain hydrogen bonds. In this review, we discuss HMW-GS, HMW-GS structure and gluten elasticity, relationship between HMW-GS and bread wheat quality and genetic engineering of the HMW-GS.

포도당 당량에 의한 탈수율과 복원율의 변화에서는 포도당 당량에 차이에 따라 탈수율은 달라졌으며 당량이 커질수록 탈수율과 복원율이 증가하는 경향을 나타내었다. 분자량에 의한 탈수율과 복원율에 변화에서는 분자량이 다른 탈수제 차이에 따라 탈수율과 복원율이 달라졌으며 분자량이 커질수록 포도당 당량과 같이 뚜렷하게 탈수율이 증가하고 복원율이 높게 나타났다. 수분함량은 동결건조한 시료가 가장 낮았으며 당량과 분자량에 따른 차이는 나타나지 않았다. 색도에서

미성숙 종자로부터 추출된 전체 RNA를 이용하여 합성한 cDNA와 LMW-GS 특이 프라이머세트를 이용하여 43개의 LMW-GS 유전자를 분리하였다. 각각의 유추 아미노산은 상동성이 높은 20개의 시그널 펩타이드, N-말단 영역, 반복서열영역 그리고 C-말단 영역을 가지며 C-말단 영역에 분자내 혹은 분자간 이황화 결합을 형성하는 전형적인 8개의 시스테인을 가지고 있었다. 이들 시스테인의 위치는 첫번째, 일곱번째를 제외하고는 보존되어 있었다. Ikeda

고지혈증 흰쥐에 있어서, 숭어(Mugil cephalus) 비늘에서 추출 제조된 저분자 콜라겐 펩타이드 추출물의 급여가 혈청 지질개선 효과 및 지질대사 이상 등에 관여하는 지질성분 및 효소의 활성 변동을 검토하기 위하여 Sprague Dawley계 흰쥐 수컷을 사용하여 본 실험을 행하였다. 기본식이만을 급여한 대조군인 BG군을 비롯한 고지혈증 유발군(CW군), 고지혈증 유발에 콜라겐 펩타이드를 섭취시킨 군(CCP군)을 5주간 실험사육하였다. 혈청

도토리 가루를 알칼리로 침전시키면서 정제하여 도토리 전분을 얻었다. 다시 이 starch를 부탄올 개량법으로 분획하여 도토리 amylose와 amylopectin을 얻었고 Corn starch와 corn amylose, corn amylopectin은 시판되는 시약을 사용하였다. 이 시료들의 분자량 분포도를 관찰하기 위하여 GCP측정을 하였다. GCP측정은 DMSO와 DMF의 용매사용과 표준물질로 pullulan을 사용하여 검정하고 각 시료들의

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the 60~% isopropanol extract of soybean(Glycine max [L.] Merr.) seed revealed two abundant proteins with molecular masses of 19 and 10 kDa. Amino acid analysis revealed that the isopropanol-extractable protein fraction was rich in cysteine. Two-dimensional gel electro-phoretic analysis indicated that the 19kDa and 10kDa proteins had pI of 4.2 and 4.0 respectively. Peptide mass fingerprints of trypsin digests of the two proteins obtained using matrix-assisted, laser desorption/ionization-time of flight (MALDI-TOF) mass spectroscopy revealed the 19kDa protein was Kunitz trypsin inhibitor and the 10kDa protein was Bowman-Birk proteinase inhibitor. When resolved under non-denaturing conditions, the isopropanol-extracted proteins inhibited trypsin and chymotrypsin activity. Results presented in this study demonstrate that isopropanol extraction of soybean seed could be used as a simple and rapid method to obtain a protein fraction enriched in Kunitz trypsin and Bowman-Birk proteinase inhibitors. Since proteinase inhibitors are rich in sulfur amino acids and are putative anticarcinogens, this rapid and inexpensive isolation procedure could facilitate efforts in nutrition and cancer research.

For the remediation of the contaminated soil with heavy metals, Cd, Cr, Cu, and Pb, the reaction parameters were optimized. Tartaric acid (TA) and oxalic acid(OA) as a washing agent were evaluated as a function of concentration, reaction time, mixing ratio of washing agent and recovery of metals. The optimum washing conditions of TA and OA were in the ratio of 1 : 20 between soil and acid solution during 2 hr reaction under unbuffered pH solutions. At the optimized reaction conditions, the removal efficiencies were compared with that of 0.1 M HC1 and ethylenediamine tetraacetic acid(EDTA). TA showed higher efficiency on the removal of Pb than that of EDTA, which established for the remediation of contaminated soil with Pb and Cd metals. The recovery of metal ions from washing solution was achieved by adding calcium hydroxide and sodium sulfide by forming the precipitation of metal hydroxide and metal sulfide. Optimum amounts of sodium sulfide and calcium hydroxide were Cd = 25g/ℓ, Cu = 5∼10g/ℓ and Pb = 5∼10g/ℓ for the washing solution of OA and 2∼5g/ℓ for the washing solution of TA, respectively. The amounts of Na2S and Ca(OH)2 for the tartaric acid was less than that of oxalic acid.

High molecular weight glutenin (HMW-Glu) subunit compositions of 73 Korean wheat cultivars and experimental lines were evaluated by using one dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This method is suitable for obtaining a good resolution of 1Dx2 and 1Ax2* without adverse effects on separation of other HMW-Glu subunits. Korean wheats examined in this study could be divided into 15 different groups on the basis of HMW-Glu subunit compositions. From the wheat lines tested, it was identified that there were three alleles at the Glu-Al, five at the Glu-Bl and three at the Glu-D1 loci. The null allele of the Glu-Al was occurred in high frequency (79.4%), while low frequencies for 1Ax1 (12.3%) and 1Ax2* (8.2%) were found. High frequency (75.3%) of the subunit pairs of 1Bx7+1By8 at the Glu-Bl loci compared with other subunits was found. The frequencies of subunits 1Dx2. 2+1Dy12 and 1Dx2+1Dy12 from the Glu-D1 loci were 54. 8% and 37.0%, respectively. However, a few Korean wheat lines (8.2%) carried 1Dx5 + 1Dy10 subunit pair which are responsible for good breadmaking quality. The information of HMW-Glu subunit compositions provide a useful tool to characterize wheat lines, and can be directly used in selection of breeding lines of different end-use properties.

The Dongbok lake water before and after alum coagulation and activated carbon adsorption were analyzed in terms of organic contents, molecular weight distributuin (MWD), and UV-absorbance. Dissolved organic compounds in the Dongbok lake were fractionated into three molecular size classes by gel permeation chromatography. The fractionation was reasonably successful in isolating compounds with MW>10,000 (Fraction Ⅰ) and those with 3,000MW (Fraction Ⅲ. The bulk of the dissolved carbon was present in compounds of molecular weight in the range of 3,000∼10,000. Alum coagulation preferentially treated molecules of high molecular weight, which has molecules larger than 10,000. The dissolved organic carbon (DOC) removal after activated carbon adsorption was high in the Fraction Ⅱ, Ⅲ. The A_260/DOC ratio after alum and activated carbon treatment was high in the Fraction Ⅰ, Ⅱ. This results suggest that the organics remaining after each treatment has a trihalomethane formation potential