• Title/Summary/Keyword: Kunitz trypsin inhibitor

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A New Soybean Cultivar "Gaechuck#1" : Black Soybean Cultivar with Lipoxygenase2,3-free, Kunitz Trypsin Inhibitor-free and Green Cotyledon

  • Chung, Jong Il
    • Korean Journal of Breeding Science
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    • v.41 no.4
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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.

A New Soybean Cultivar "Gaechuck#2": Yellow Soybean Cultivar with Lipoxygenase2,3-free and Kunitz Trypsin Inhibitor-free

  • Chung, Jong Il
    • Korean Journal of Breeding Science
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    • v.41 no.4
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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.

Selection of Lipoxygenase, Kunitz Trypsin inhibitor and 7Sα′-subunit Protein Free Soybean Strain (Lipoxygenase, Kunitz Trypsin inhibitor, 7Sα′-subunit 단백질이 결핍된 콩 계통의 선발)

  • Sung, Mi-Kyung;Kim, Kyung-Roc;Park, Jung-Soo;Han, Eun-Hui;Nam, Jin-Woo;Chung, Jong-Il
    • Journal of agriculture & life science
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    • v.44 no.5
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    • pp.29-33
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    • 2010
  • Soybean (Glycine max (L.) Merr.) seed is the main source of protein and oil for human and animal. The use of soybean protein has been expanded in the food industry due to their excellent nutritional benefits. But, antinutritional and allergenic factors are exist in the raw mature soybean. Lipoxygenase, Kunitz trypsin inhibitor (KTI) protein, and ${\alpha}^{\prime}-subunit$ of 7S globulin are main antinutritional factors in soybean seed. Breeding of a new soybean strain with lacking these components is needed. The objective of this research was to select new soybean line with lipoxygenase-free, KTI-free, and ${\alpha}^{\prime}-subunit$ free (lx1lx1lx2lx2lx3lx3titicgy1cgy1 genotype). Total 434 $F_2$seeds were obtained from the cross of cultivar, "Gaechuck#2" and PI506876. Presence and absence of lipoxygenase, KTI protein, and ${\alpha}^{\prime}-subunit$ of 7S globulin was tested by SDS electrophoresis using a partial seed of each $F_2$seed. Only one $F_2$seed with lacking these three components was selected and was planted to $F_2$plant. Absence of lipoxygenase, KTI, and ${\alpha}^{\prime}-subunit$ protein was confirmed on the $F_3$seeds harvested. Selected line with lx1lx1lx2lx2lx3lx3titicgy1cgy1 genotype might be useful for soybean breeding.

Identification of the Precursor for the Soybean Kunitz Trypsin Inhibitor (대두 Kunitz Trypsin Inhibitor 전구체의 동정)

  • Kim, Chung-Ho;Kim, Su-Il;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.32 no.3
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    • pp.222-231
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    • 1989
  • Three classes of proteinase inhibitors are known in soybean; the Kunitz trypsin inhibitor (SKTI), the Bowman-Birk proteinase inhibitor and its isoinhibitors. To study the molecular structure and expression characteristics of the SKTI, antibody was obtained by immunizing rabbit with the SKTI purified from soybean by preparative electrophoresis. Anti-SKTI antibody was not only specific for mature SKTI in soybean seed but also recognized the precursor which was synthesized in vitro. Translation in vitro was carried out in wheat germ extract with polyadenylated mRNA isolated from developing soybean seeds. One of the seed specific translation products, MW 24K, was identified to be the precursor for the SKTI by immunoprecipitation with anti-SKTI antibody. Mature SKTI of MW 20K, however, was not detected in the translates in vitro. These results suggest that the precursor polypeptide is synthesized from the mRNA and is cleaved to yield mature SKTI in soybean seed. The SKTI gene was expressed with the maturation of soybean seed in a tissue-specific and development stage-specific manner.

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A Simple and Rapid Method to Isolate Low Molecular Weight Proteinase Inhibitors from Soybean

  • Krishnan Bari B.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.4
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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.

Identification of Molecular Markers Linked to Ti Locus in Soybean

  • Kim Myung Sik;Park Min Jung;Hwang Jung Gyu;Jo Soo Ho;Ko Mi Suk;Chung Jong Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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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.

Seed Transmission Rates of Bean pod mottle virus and Soybean mosaic virus in Soybean May Be Affected by Mixed Infection or Expression of the Kunitz Trypsin Inhibitor

  • Nam, Moon;Bae, Hanhong;Hammond, John;Domier, Leslie L.;Youn, Young-Nam;Lee, Bong-Choon;Lim, Hyoun-Sub
    • Research in Plant Disease
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    • v.19 no.2
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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.

Zika Virus: Discovering Effective Protease Inhibitors via Template-Based Modeling

  • Choi, Yongseok
    • Proceeding of EDISON Challenge
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    • 2016.03a
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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.

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Germplasm Detection for titi Genotype Using SSR Marker in Soybean

  • Kim, Myung-Sik;Jeong, Woo-Hyeun;Nam, Ki-Chul;Park, Mo-Se;Lee, Kyoung-Ja;Chung, Jong-Il
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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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.

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Independent Inheritance between df2 gene and ti gene in Soybean

  • Han, Eun-Hui;Sung, Mi-Kyung;Kim, Kyung-Roc;Park, Jung-Soo;Nam, Jin-Woo;Chung, Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.1
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    • pp.14-17
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    • 2011
  • Dwarfuess and Kunitz trypsin inhibitor (KTI) protein in soybean is useful traits for basic studies. df2 and ti gene control dwarfness and the expression of Kunitz trypsin inhibitor (KTI) protein in soybean, respectively. The objective of this research was to verify genetic linkage or independent inheritance of df2 and ti loci in soybean. The $F_2$ population was made by cross combination between "Gaechuck#2" (Df2Df2titi genotype, KTI protein absence and a normal growth type) and T210 (df2df2TiTi genotype, a dwarf growth type and KTI protein present). A total of 258 $F_2$ seeds were analyzed for the segregation of KTI protein using SDS-PAGE. And so, 198 $F_2$ plants were recorded for the segregation of dwarfness. The segregation ratio of 3 : 1 for Ti locus (201 Ti_ : 57 titi) and Df2 locus (143 Df2_ : 55 df2df2) was observed. Segregation ratio of 9 : 3 : 3 : 1 (116 Ti_Df2_: 44 Ti_df2df2: 27 titiDf2_: 11 titidf2df2) between df2 gene and ti gene was observed ($x^2$=3.53, P = 0.223). These results showed that df2 gene was inherited independently with the ti gene in soybean.