• 한국육종학회지
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• 제41권4호
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• pp.603-606
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• 2009

Lipoxygenase and Kunitz trypsin inhibitor protein of mature soybean [Glycine max (L.) Merr.] seed are main anti-nutritional factors in soybean seed. A new soybean cultivar, "Gaechuck#1" with the traits of black seed coat, green cotyledon, lipoxygenase2,3 and Kunitz trypsin inhibitor protein free was developed. It was selected from the population derived the cross of "Gyeongsang#1" and C242. Plants of "Gaechuck#1" have a determinate growth habit with purple flowers, brown pubescence, black seed coat, black hilum, oval leaflet shape and brown pods at maturity. Seed protein and oil content on dry weight basis have averaged 39.1% and 16.2%, respectively. It has shown resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and soybean pod borer. "Gaechuck#1" matured on 5-10 October with a plant height of 50 cm. The 100-seed weight of "Gaechuck#1" was 23.2g. Yield of "Gaechuck#1" was averaged 2.2 ton/ha from 2005 to 2007.

• 한국육종학회지
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• 제41권4호
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• pp.612-615
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• 2009

Lipoxygenase and Kunitz trypsin inhibitor protein are the main antinutritional factor in mature soybean seed. A new soybean cultivar, "Gaechuck#2" with yellow seed coat, lipoxygenase2,3-free and Kunitz trypsin inhibitor protein-free was developed. It was selected from the population derived from the cross between "Jinpumkong2ho" and C242. Plants of "Gaechuck#2" have determinate growth habit with purple flowers, tawny pubescence, yellow seed coat, yellow hilum, oval leaflet shape and brown pods at maturity. Seed protein and oil content on a dry weight basis were 40.7% and 18.7%, respectively. It has shown a resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and soybean pod borer. Gaechuck#2 matured in 4 October with plant height of 54cm and a 100-seed weight of 24.4g. Average Yield of Gaechuck#2 was 230 - 250 kg/10a in 2005 - 2007.

• 농업생명과학연구
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• 제44권5호
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• pp.29-33
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• 2010

콩은 식물성 단백질 및 지방의 주요 공급원이고 콩 종실에는 기능성 성분이 많이 함유되어져 있어 소비가 점차 증가하고 있지만 Lipoxygenase, Kunitz Trypsin inhibitor, $7S{\alpha}^{\prime}-subunit$ 단백질과 같은 성분들이 존재하는데 이는 품질과 영양가치를 떨어뜨리고 섭취시 알러지를 일으키기도 한다. 콩에서 유전적으로 이러한 성분이 결핍되어져 있는 유전자형의 선발은 품질이 우수한 콩 육종의 기초단계이다. "개척2호"와 PI506876의 교배로부터 434개의 $F_2$종자를 얻어 $F_2$종자의 일부를 사용하여 SDS-PAGE로 각각의 종자를 분석한 결과 Lipoxygenase와 Kunitz Trypsin inhibitor 및 7S의 ${\alpha}^{\prime}-subunit$ 단백질이 모두 결핍되어져 있는 lx1lx1lx2lx2lx3lx3titicgy1cgy1 유전자형을 가진 종자를 선발하여 $F_2$식물체로 길러 성숙 후 $F_3$종자를 수확하였다. $F_3$종자로부터 Lipoxygenase, Kunitz Trypsin inhibitor, $7S{\alpha}^{\prime}-subunit$ 단백질이 모두 결핍되어져 있음을 재확인하였으며 선발된 종자는 고품질 콩 품종 육성에 유용하게 활용될 것으로 기대된다.

• Applied Biological Chemistry
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• 제32권3호
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• pp.222-231
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• 1989

대두에 존재하는 단백질 분해 효소 저해제로는 Kunitz trypsin inhibitor (SKTI), Bowman-Birk proteinase inhibitor, 그리고 그의 isoinhibitor들이 알려져 있다. SKTI는 분자량이 20K이며 181개의 아미노산으로 구성되어 있다. SKTI의 분자 구조 및 발현 특이성을 밝히기 위하여, 콩에서 순수 분리한 SKTI를 항원으로 사용하여 항체를 제조하였다. 이 항체는 항원으로 사용한 SKTI에 대하여 특이적으로 반응할 뿐만 아니라 기내에서 합성된 그의 전구체와도 특이적으로 반응하였다. 기내에서의 단백질 합성은 미성숙 대두 종자에서 mRNA를 분리한 후, wheat germ extract를 이용하여 실시하였다. 합성된 단백질 중에서 대두에 존재하는 SKTI는 검출되지 않는 반면에 면역침전법에 의해 분자량이 24K인 전구체가 존재하고 있음을 확인하였다. 이와 같이 SKTI는 mRNA로부터 전구체가 합성된 후 post-translational modification에 의해 완전한 SKTI로 변환됨을 알 수 있었다. 또한 SKTI는 대두 종자의 성숙과 함께 발현이 되는 것으로 보아 조직 및 분화 특이성 발현형태를 나타내는 것을 알 수 있었다.

• 한국작물학회지
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• 제49권4호
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• pp.342-348
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• 2004

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.

• 한국작물학회지
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• 제49권5호
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• pp.419-422
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• 2004

Soybean is a major source of protein meal in the world. Kunitz trypsin inhibitor (KTI) protein is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The objective of this research was to identify RAPD markers linked to KTI protein allele using bulked segregant analysis. Cultivar Jinpumkong2 (TiTi) was crossed with C242 (titi, absence of KTI protein) and F. seeds were planted. The $F_1$. plants were grown in the greenhouse to produce $F_2$ seeds. Each $F_2$ seed from $F_1$. plants was analysed electrophoretically to determine the presence of the KTI protein band. The present and absent bulks contained twenty individuals each, which were selected on the basis of the KTI protein electrophoresis, respectively. Total 94 $F_2$ individuals were constructed and 1,000 Operon random primers were used to identify RAPD primers linked to the Ti locus. The presence of KTI protein is dominant to the lack of a KTI protein and Kunitz trypsin inhibit protein band is controlled by a single locus. Four RAPD primers (OPAC12, OPAR15, OPO12, and OPC08) were linked to the Ti locus. RAPD primer OPO12 was linked to Ti locus, controlling kunitz trypsin inhibitor protein at a distance of 16.0 cM. This results may assist in study of developing fine map including Ti locus in soybean.

• 식물병연구
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• 제19권2호
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• pp.114-117
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• 2013

To facilitate their spread, plant viruses have developed several methods for dispersal including insect and seed transmission. While insect transmission requires virus stability against insect digestion, seed-transmitted viruses have to overcome barriers to entry into embryos. Bean pod mottle virus (BPMV) is transmitted through seed at levels typically below 0.1%, but co-infection with Soybean mosaic virus (SMV) enhanced the seed transmission rate of BPMV in one experiment. In contrast, the rate of SMV seed transmission was not affected by BPMV co-infection. In a second preliminary study, the rate of SMV transmission was lower in an isoline of Williams 82 that contained a null mutation for the Kunitz trypsin inhibitor gene than in Williams 82. In this preliminary study, we observed that factors such as protease inhibitor expression and dual infection may affect the frequency of seed transmission of BPMV and SMV.

• EDISON SW 활용 경진대회 논문집
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• 제5회(2016년)
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• pp.86-89
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• 2016

The Zika virus, which is a member of the flavivirus genus, poses a serious threat to humanity because there is no vaccine or cure. Zika is suspected to cause microcephaly, and it is rapidly spreading throughout parts of Brazil. Surprisingly, there are no known protein structures for the virus which are essential for drug and vaccine development. This paper investigates the Zika virus's nonstructural proteins with template-based modeling by using GalaxyTBM/Refine/SC. GalaxyDock was used to examine the effectiveness of various known serine protease inhibitors in inhibiting the Zika viral protease. In testing five inhibitors, Kunitz soybean trypsin inhibitor showed the strongest binding affinity (-10.082 kcal/mol). This paper provides a rudimentary foundation for further drug discovery research regarding the Zika virus.

• 생명과학회지
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• 제12권2호
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• pp.151-157
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• 2002

한방재료의 하나인 택사(Alismatis Rhizoma, AR)로부터 단백성 trypsin inhibitor(TI)를 분리, 정제하여 특성을 조사하였다. 정제과정은 0-80.% 포화 황산암모늄을 이용한 염석법, DEAE-cellulose ion exchange chromatography, Sep-hadex G-150 chromatography 등을 이용하였다. 정제된 ARTI의 분자량을 gel filtration과 SDS-PAGE 한 결과 모두 약 23,000 Da으로 나타나 monomer로 되어 있는 것으로 나타났다. 온도안정성에 있어 0-6$0^{\circ}C$에서는 안정하였으나 그 이상의 온도에서는 약 35%가지 안정성이 떨어졌다. ARTI와 상품화된 soybean kunitz inhibitor의 저해능을 비교해 본 결과 ARTI 및 soybean inhibitor 각각의 농도가 0.071 $\mu$M, 1.7 $\mu$M일 때 0.025 g/$m\ell$ trypsin활성을 50% 정도 저해하는 것으로 나타났다. ARTI의 trypsin의 가수분해반응에 대한 저해형태는 비경쟁적 저해형인 것으로 나타났으며 km값은 0.81 $\mu$M이었다.

• Journal of Crop Science and Biotechnology
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• 제10권3호
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• pp.159-162
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• 2007

Soybean Kunitz trypsin inhibitor(SKTI) protein is a small, monomeric and non-glycosylated protein containing 181 amino acid residues and is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The objective of this research is to confirm SSR marker(Satt228) tightly linked to the Ti locus using several germplasm accessions with TiTi or titi genotypes for MAS in soybean breeding programs. TiTi genotypes('Jinpumkong2', 'Clark', and 'William') had allele1 and titi genotypes(PI196168, C242, W60, and PI157440) had allele2 in Satt228 marker analysis. 'Jinpumkong2', 'Clark', and 'William'(TiTi genotype) had a Kunitz trypsin inhibitor protein of 21.5 kDa size, and PI196168, C242, W60, and PI157440(titi genotype) did not have the band in protein gel electrophoresis from the mature seed. Cosegregation between the SKTI protein(21.5 kDa size) and allele of Satt228 marker was observed in seven germplasm accessions with different genetic backgrounds. Any recombination between the SKTI protein and allele of the Satt228 marker was not observed. This result indicates that Satt228 marker may effectively utilized to select the plants with the titi genotype.