• 한국식품영양학회지
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• 제12권2호
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• pp.184-190
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• 1999

Soybean protein consists of two major components $\beta$-conglycinin and glycinin which together consti-tute 70% of the total seed storage protein at maturity. $\beta$-Conglycinin is trimeric glycoprotein and for-med by the assembly of various combinations of three subunits $\alpha$,$\alpha$' and $\beta$ which have molecular weig-hts of 69,000, 72,000 and 42,000, respectively. Recently $\beta$-conglycinin was identified as powerful LDL lip-oprotein receptor activation hypercholesterolemia and major allergenic proteins. To investigate these reasons we constructed an expression system of cDNA encoding $\alpha$-subunit of $\beta$-conglycinin in Escherichia coli and purified the expressed protein. The pro-$\beta$-conglycinin synthesized in Escherichia coli BL 21 (DE3)comprised approximately 15% of the total bacterial proteins and the expressed protein are formed sol-uble and trimer such as native protein in Escherichia coli cells. The highly expressed protein was purified to homogeneity by salt precipitation with 20~40 % ammonium sulfate ion-exchange chromatography with Q-sepharose and hydrophobic column chromatography with Butyltoyopearl.

• Food Science and Biotechnology
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• 제14권1호
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• pp.11-15
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• 2005

Dynamic shear moduli of isolated soy protein solutions upon heating were measured to monitor gelation. Onsets of gelation coincide with onset temperatures of denaturation in glycinin and ${\beta}$-conglycinin solutions, whereas in isolated soy proteins, onset of gelation was above denaturation temperature of ${\beta}$-conglycinin with storage modulus increasing in two steps. The first increase in storage modulus of isolated soy proteins occurred at about $78.5^{\circ}C$, while the second increase started at about $93^{\circ}C$. Gel properties of soy protein gels having different proportions of glycinin and ${\beta}$-conglycinin were measured by compression-decompression test. ${\beta}$-conglycinin was responsible for gel elasticity. Glycinin significantly increased hardness, toughness, and fracturability of gels at high heating temperature near $100^{\circ}C$. Results reveal texture of soy protein gels can be controlled by regulating ratio of glycinin to ${\beta}$-conglycinin and heating temperature.

• 한국작물학회지
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• 제47권2호
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• pp.85-94
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• 2002

Antibodies raised against the purified p-subunit of $\beta$-conglycinin were used in immunohistochemical studies to monitor the pattern of $\beta$-conglycinin mobilization in the cotyledons during soybean [Glycine max (L.) Merr.] seed germination. Western blot analysis revealed that the break down of the $\beta$-subunit of $\beta$-conglycinin commenced as early as 2 days after seed imbibition (DAI). Concurrent with the degradation of the $\beta$-subunit of $\beta$-conglycinin, accumulation of 48, 28, and 26 kD proteolytic intermediates was observed from 2 to 6 DAI. Western blot analysis also revealed that the acidic subunit of glycinin was mobilized earlier than the basic subunit. The basic glycinin subunit was subjected to proteolysis within 2 DAI resulting in the appearance of an intermediate product approximately 2 kD smaller than the native basic glycinin subunit. In contrast to the major seed storage proteins, lipoxygenase was subjected to limited proteolysis and was detected even after 8 DAI. The first sign of $\beta$-conglycinin breakdown was observed near the vascular strands and proceeded from the vascular strands towards the epidermis. Protein A-gold localization studies using thin sections of soybean cotyledons and antibodies raised against the $\beta$-subunit of $\beta$-conglycinin revealed intense labeling over protein bodies. A pronounced decrease in the protein A-gold labeling intensity over protein bodies was observed at later stages of seed germination. The protein bodies, which were converted into a large central vacuole by 8 DAI, contained very little 7S protein as evidenced by sparse protein A-gold labeling in the vacuoles.

• Applied Biological Chemistry
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• 제36권6호
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• pp.547-552
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• 1993

대두 종자 저장 단백질의 일종인 ${\beta}-conglycinin$ ${\alpha}'$ subunit 유전자의 upstream 지역에 결합하여 전사 조절에 관여하리라 추정되는 대두 핵의 DNA 결합 단백질을 조사하기 위하여 대두 핵 추출물과 S-100을 조제하였다. 염기서열이 AACCCA-27 bp-AACCCA인 합성 DNA를 pUC19에 클로닝한 플라스미드 pSE3를 EcoRI과 HindIII로 절단하여 절편을 분리하고 $^{32}P$ 로 표지하여 이를 gel mobility shift assay 탐침으로 이용한 결과, 대두 핵의 DNA 결합 단백질의 일종인 SEF3(soybean embryo factor 3)의 역가가 핵 추출물과 S-100에서 검출되었다. 각각 CATGCAT, AACACA 염기 서열을 가지는 DNA를 탐침으로 이용하여 SEF3 이외의 DNA 결합 단백질의 역가를 조사한 결과 대두 핵 추출물과 S-100에서 각기의 염기 서열에 결합하는 수 종의 DNA 결합 단백질이 확인되었으나 두 시료에서 공통된 양상을 보여주는 DNA 결합 단백질의 역가는 확인되지 않았다. 또한 대두 S-100의 경우에는 개화 후 32일 부근에 SEF3 역가가 검출되는 데에 비하여, 핵 추출물에는 20일 전후에 SEF3 역가가 나타나서 32일 부근에 역가가 현저히 증가하였다.

• Applied Biological Chemistry
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• 제36권6호
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• pp.553-556
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• 1993

대두 종자 저장 단백질의 일종인 ${\beta}-conglycinin$의 ${\alpha}'$subunit 유전자 upstream 영역에 결합하여 전사 조절에 관여하리라 추정되는 SEF3(soybean embryo factor 3)의 발현을 조사하기 위하여 대두 핵 추출물을 조제하였다. 두개의 AACCCA를 포함하는 SE3 DNA를 $^{32}P$로 표지한 후 gel mobility shift assay 탐침으로 이용하여 대두 발달 과정 중의 SEF3의 역가를 조사하여 본 결과, 개화 후 16일부터 32일까지의 역가는 증가하나 SE3-SEF3 결합체 이동도는 감소하였다. 핵 추출물을 alkaline phosphatase 처리하면 결합체의 이동도가 다시 증가하였으나 이 현상은 phosphate에 의해 저해되었다. 그리고 결합체의 형성은 반응 pH 6.8과8.5사이에서는 큰 영향을 받지 않았다.

• 한국자원식물학회지
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• 제10권3호
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• pp.221-226
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• 1997

Improvement of seed protein quality might be an essential issus in soybean and would give more profit directly to both farmers and users. This study was carried out to investigate the effects of reduced-S form(s) on seed storage protein components in soybean during seed filling stages. The reduced-S forms during seed fill were sodium thiosulfate, sodium sulfite, sodium sulfide, thioaceteat, $\beta$-mercaptoethanol, thiourea, thiamine-HCI, L-cysteine, L-cystine, and L-methionine. Seed storage protein concentration did not appear to be affected by any reduced-S forms. However, glycinin and $\beta$-conglycinin concentration seemed to be changed greatly by L-methionine. This resulted in the increase in the 11S/7S ratio(3.58). Among the $\beta$-conglycinin, $\beta$-subunit was not accumulated at all. $\alpha$-subunit concentration appeared to be decreased and $\alpha'$-subunit concentration was not altered in comparison with sulfate control. Also, $\beta$-conglycine concentration, especially $\beta$-subunit concentration, tended to be decreased with L-cystine treatment, resulting in an increase in the 11S/7S ratio(1.83). The glycinin concentration tended to be increased at the expense of the decrease in the $\beta$-conglycinin concentration. Therefore, it is suggested that enhancing soybean protein quality would be achieved by improving metabolic pathways of S assimilation in soybean plants during seed filling period under sulfate-sufficient condition.

• 한국식품영양학회지
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• 제9권4호
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• pp.404-408
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• 1996

콩 단백질은 글리시닌과 $\beta$-콘글리시닌을 주요 성분으로 한다. 영양성 및 가공 특성을 개선하기 위하여 유전자공학적인 방법을 시도하였다. 즉 $\beta$-콘글리시닌의 $\beta$-서브유니트를 유전자 클로닝하고 대장균에서 발현시켰다. 발현벡타는 pET 21d이며 플라스미드를 구축하여 E. coli BL21(DE3)에 형질전환 시켰다. 발현된 단백질은 균체 전체 단백질의 20%이며 가용화 상태로 축적되었다. 축적 발현단백질은 천연의 $\beta$-콘글리시닌과 동일항 트리머로 확인되었다. 정제는 황산암모늄 20~40% 분별침전, Q-Sepharose 이온교환크로마토그라피, Butyltoyopearl 소수성 컬럼크로마토그라피로 하였다. 이것은 콩 단백질의 특성을 규명하는데 필요한 대장균 대량 발현계를 확립하고 발현 단백질의 정제방법을 확립한 결과이다.

• Applied Biological Chemistry
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• 제38권1호
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• pp.63-66
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• 1995

본 실험에서는 대두 ${\beta}-conglycinin$의 ${\alpha}'$ subunit 유전자의 발현에 관여하리라 추정되는 대두 배인자 3(SEF3) 역가를 가지는 대두 핵 추출물 조제 방법에 대하여 조사하였다. 역가 조사를 위한 DNA의 표지 반응 후에 추가로 1 mM dATP를 첨가하여 반응을 지속한 경우가 첨가하지 않은 경우보다 약 4배 높은 방사능 역가의 DNA 탐침을 얻을 수 있었다. 이러한 DNA 탐침을 이용하여 핵 추출물 조제 방법에 따른 대두 배인자 3의 역가를 gel mobility shift assay로 조사한 결과, 조제 중 glycerol 첨가가 배인자 3을 안정화시켰고, polyethylenimine을 이용한 대두 핵 추출물 조제법이 배인자 3의 비역가를 증가시키는 데 이용될 수 있음을 알 수 있었다.

• Food Science and Biotechnology
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• 제15권1호
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• pp.128-132
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• 2006

This study was undertaken to investigate the effect of peptic and chymotryptic hydrolyses of 7S globulin, the major allergen of soybean protein, on its allergenicity, as measured by enzyme linked immunosorbent assay (ELISA), and to identify the allergenic hydrolyzed fragments of 7S globulin using SDS-PAGE. When 7S globulin was hydrolyzed by pepsin, the allergenicity was reduced by over 50%. However, the allergenicity of 7S globulin reduced by peptic hydrolysis was recovered in the sera from 5 out of 10 patients following sequential chymotryptic hydrolysis. Two fragments, with molecular weights 20-25 and 13-16 kDa, among the hydrolysate of 7S globulin by sequential pepsin and chymotrypsin showed reactivity with sera from 10 soybean-allergenic patients. As a result of the theoretical hydrolyses of ${\beta}$-conglycinin, which is a major protein of 7S globulin, it is suggested that the 20-25 kDa fragments were the fragments of the ${\alpha}$-subunit of ${\beta}$'-conglycinin and that the 10-16 kDa fragments were from the ${\alpha}$'-subunit.

• 농업생명과학연구
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• 제43권5호
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• pp.39-42
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• 2009

인간과 가축의 영양을 위한 식물성 단백질의 주요 공급원은 콩이며 콩 단백질은 영양 및 기능성면에서 우수하여 소비가 점차 증가하고 있다. 그러나 콩 단백질에는 알러지를 일으키고 영양가치를 떨어뜨리는 성분도 포함되어져 있다. 7S 및 11S 글로블린은 콩 저장단백질의 대부분을 차지하며 7S는 영양가치가 떨어지고 7S의 함량을 줄어든 콩 계통 육성에 대한 관심이 높아지고 있다. 7S 성분중의 하나인 ${\alpha}^{\prime}$-subunit의 유전양상을 파악하기 위하여 진품콩2호와 PI506876의 교배로부터 98개의 F2 종자가 얻어졌다. SDS-PAGE로 각각의 종자를 분석한 결과 ${\alpha}^{\prime}$-subunit을 가진 종자가 70개였고 결핍된 종자가 28개였다. 이러한 유전양상은 단인자 유전원칙 (${\chi}^2=0.667$, P=0.414)과 일치하여 콩 종자에서 7S의 ${\alpha}^{\prime}$-subunit 단백질은 한 개의 유전자에 의 해서 좌우되었다. 이 결과는 7S 단백질 함량이 줄어든 콩 계통 선발에 유용하게 활용될 것으로 기대된다.