• Title/Summary/Keyword: Kunitz trypsin inhibitor protein

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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 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.

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.

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.

The Development of a New Soybean Strain Without Kunitz Trypsin Inhibitor, Lectin, and 7S α' Subunit Protein (쿠니츠트립인히비터, 렉틴 및 7S α' 서버유닛 3가지 단백질이 없는 콩 계통의 개발)

  • Chae, Won Gi;Choi, Sang Woo;Kang, Gyung Young;Chung, Jong Il
    • Journal of Life Science
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    • v.30 no.7
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    • pp.592-597
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    • 2020
  • Soybean [Glycine max (L.) Merr.] seeds contain an average of 40% protein on a dry weight basis, but they also contain antinutritional elements such as lectin, Kunitz trypsin inhibitor (KTI), and 7S α'- subunit protein. The objective of this research was to develop a new soybean genotype with triple recessive alleles for these elements. Three parents (Gaechuck#2, PI506876, and Le-16) were used to develop the genetic population, and the presence of lectin and KTI protein was detected using Western blot while 7S α' subunit protein was detected using SDS-PAGE. One F3 plant strain with proper agronomical traits such as type, height, seed quality, and 100-seed weight was selected. The genotype of the developed strain is titilelecgy1cgy1, that is KTI, lectin, and 7S α' subunit protein free. The new strain has a purple flower, determinate growth habit, and light yellow pods at maturity. The seed has a buffer hilum and is yellow in color. The new strain's height was 58 cm compared to the Daewonkong cultivar at 46 cm, and its 100-seed weight was 27.1 g, smaller than the Daewonkong at 29.0 g. This is the first new soybean strain with the titilelecgy1cgy1 genotype, and it can be used to improve yellow soybean cultivars of high quality and function.

Selection of rs2rs2titi Soybean Genotype with Yellow Seed Coat (rs2rs2titi 유전자형을 가진 노란 콩 계통 선발)

  • Choi, Sang Woo;Park, Jun Hyun;Chung, Jong Il
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1285-1289
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    • 2018
  • Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrates, isoflavones, and other nutrients for humans and animals. But, antinutritional factors in the raw mature soybean are exist. Kunitz trypsin inhibitor (KTI) protein and stachyose are main antinutritional factors in soybean seed. The genetic removal of the antinutritional factors will improve the nutritional value of soybean seed. The objective of this research was to breed a new yellow soybean strains (rs2rs2titi genotype) with the traits of lacking of KTI protein and low content of stachyose. Breeding population was developed from the cross of "Jinyangkong" and 15G1 parents. Presence or absence of KTI protein was detected based on Western Blot technique. Content of stachyose in mature seed was detected by HPLC. Total four new strains (603-1, 603-2, 625, and 694) with KTI protein free and low content of stachyose were selected. Four strains (603-1, 603-2, 625, and 694) have yellow seed coat and hilum. Plant height of 603-1 strain was 65 cm and 100-seed weight was 29.2 g. Plant height of 603-2 strain was 66 cm and 100-seed weight was 26.2 g. Plant height of 625 strain was 64 cm and 100-seed weight was 27.1 g. Content of stachyose for four new strains was 3.0~3.50 g/kg. Four strains selected in this research will be used to improve new yellow soybean cultivar with KTI protein free, and low content of stachyose.

Physicochemical Characteristics and Antioxidant Activity of Kanjang made from Soybean Cultivars Lacking Lipoxygenase and Kunitz Trypsin Inhibitor Protein (Lipoxygenase와 Kunitz Trypsin Inhibitor 단백질 결핍콩으로 제조한 간장의 이화학적 특성 및 항산화 활성)

  • Hwang, Cho-Rong;Lee, Soo-Jung;Kang, Jae-Ran;Kwon, Min-Hye;Kwon, Hyo-Jin;Chung, Jong-Il;Sung, Nak-Ju
    • Journal of agriculture & life science
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    • v.46 no.5
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    • pp.111-125
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    • 2012
  • In order to evaluate suitability by processing for non-GM soybean cultivars such as Gaechuck#2, Jinyangkong and CJ#1 lacking lipoxygenase (LOX) and kunitz trypsin inhibitor (KTI) protein, physicochemical characteristics and antioxidant activity of Kanjang made from above soybean were compared to Kanjang made from a conventional cultivar (Taekwangkong). Proximate compositions of soybeans were similar for the 4 kinds cultivars. Total phenol and flavonoid contents were significantly higher in cultivars lacking LOX and KTI protein than the Taekwangkong. In Kanjang, contents of total and reducing sugar were higher in Taekwangkong Kanjang than Kanjang from made cultivars lacking LOX and KTI protein. Contents of total and amino type nitrogen were the highest in the Jinyangkong Kanjang. Mineral contents were higher in the Jinyangkong and CJ#1 Kanjangs, amino acid contents were higher in the Kanjang made from 3 cultivars lacking LOX and KTI protein than the Taekwangkong. Taste of the Jinyangkong Kanjang with higher sweety and savory was also found to be superior to that of others in overall acceptability evaluation. Total phenol and flavonoid contents in Kanjang were significantly higher in the Kanjang made from cultivars lacking LOX and KTI protein than the Taekwangkong. Radical scavenging activity of Kanjang was increased in the total phenol contents dependent on. Reducing power by ferric-reducing antioxidant potential (FRAP) was significantly higher the Kanjang made from Gaechuck#2 and CJ#1 than the Taekwangkong. $Fe^{2+}$ chelating activity was higher in Taekwangkong Kanjang than the other cultivars, but its activity was similar to Jinyangkong Kanjang. Therefore, higher nutritional composition, total phenol and flavonoids contents and antioxidant activity in the Kanjang made from soybean cultivars lacking LOX and KTI protein might be provide better benefit for manufacture of another their products.