• Title/Summary/Keyword: trypsin inhibitor.

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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.

Tempeh Fermentation from a Mixture of Soybean and Sorghum Grain (대두-수수 혼합곡물의 템페발효)

  • Kim, Chong-Tai;Kim, Chul-Jin;Kim, Dong-Chul;Kwon, Tae-Won
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.668-674
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    • 1990
  • Tempeh-type fermented products were prepared from soybean, sorghum or mixture of soybean and sorghum(1 : 1) with the traditional Indonesian inoculum(LARU : mixed cultures of Rhizopus oligosporus). Fermentation increased protein and fiber contents in the soybean tempeh(ST) and tempeh of soybean-sorghum mixture(SSM). Fat content was slightly higher in sorghum tempeh(SGT) and SSM than that of control. During the fermentation, pH, soluble solid and soluble nitrogen were increased, while no significant change was found in the total solids. The trypsin inhibitor activity(TIA) and phytic acid content decreased after 32 hrs fermentation. It is suggested that Rhizopus oligosporus is capable of hydrolyzing trypsin inhibitor and phytic acid of the substrate. Thiamine and niacin contents increased in all samples as compared with the unfermented control. In amino acid level, there were some decreased in total amino acids after 32 hrs fermentation in three types of tempeh. While the concentrations of lysine, valine. tyrosine and alanine in ST, SGT and SSM were increased those of serine and glutamic acid were decreased in compare to the unfermented control.

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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.

Effect of Cultivars, Cooking and Processing on the Trypsin Inhibitor Activity of Soybean

  • Felipe, Penelope;Yang, Yoon-Hyung;Lee, Jung-Hee;Sok, Dai-Eun;Kim, Hyoung-Chin;Yoon, Won-Kee;Kim, Hwan-Mook;Kim, Mee-Ree
    • Preventive Nutrition and Food Science
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    • v.10 no.1
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    • pp.6-10
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    • 2005
  • The trypsin inhibitor activity (TIA) of various soybean cultivars was evaluated by measuring the inhibition of trypsin activity using N-benzoyl-DL-arginine-p-nitro-anilide (BAPNA) as the substrate. The TIA values of eleven white shelled soybean cultivars including a glyphosate-tolerant soybean (16.58 to 17.90㎎/g) were not significantly different among cultivars. Black shelled soybeans had higher TIA values, ranging from 40.09 to 52.11㎎/g, compared to white shelled soybeans (p<0.05). When the TIA of commercially processed soybean foods were determined, no TIA was detected in soysauce, tofu and soybean paste. During conventional moist heating, the IT/sub 50/ (Time required to reach 50% inhibition of TIA) values were decreased as heating temperature and cooking pressure increased. The IT/sub 50/ values of moist heating were estimated to be 91.68, 37.71 and 19.50 min at 60, 80 and 100℃, respectively. The IT/sub 50/ value of microwave cooking was 4.75 min at medium heat, while that of the pressure cooking at 120℃ was only 2.62min. Moreover, there was a negative relationship between temperature and IT/sub 50/ values (R=0.92, p<0.01). The TIA of soybean sprouts was completely inactivated after heating at 100℃ for 5 min, although fresh soybean sprouts showed one fifth of the TIA value of white shelled soybeans. Based on our results, pressure cooking is the most effective cooking method to reduce TIA in soybeans.

Streptomyces griseus HH1, An A-factor Deficient Mutant Produces Diminished Level of Trypsin and Increased Level of Metalloproteases

  • Kim, Jung-Mee;Hong, Soon-Kwang
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.160-168
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    • 2000
  • A-factor I a microbial hormone that can positively control cell differentiation leading to spore formation and secondary metabolite formation in Streptomyces griseus. to identify a protease that is deeply involved in the morphological and physiological differentiation of Streptomyces, the proteases produced by Streptomyces griseus IFO 13350 and its A-factor deficient mutant strain, Streptomyces griseus HH1, as well as Streptomyces griseus HH1 transformed with the afsA gene were sturdied. In general Streptomyces griseus showed a higher degree of cell growth and protease activity in proportion to its ability to produce a higher amount of A-factor. In particular, the specific activity of the trypsin of Streptomyces griseus IFO 13350 was greatly enhanced more than twice compared with that of Streptomyces griseus HH1 in the later stage of growth. The specific activity of the metalloprotease of Streptomyces griseus HH1 was greatly enhanced more than twice compared with that of Streptomyces griseus IFO 13350, and this observation was reversed in the presence of thiostreptione, However, Streptomyces griseus HH1 transformed with the afsA gene showed a significantly decreased level of trypsin and metalloprotease activity compared with that of the HH1 strain. There was no significant difference between Streptomyces griseus IFO 13350 and HH1 strain in their chymotrypsin and thiol protease activity, yet the level of leu-amionpeptidase activity was 2 times higher in Streptomyces griseus HH1 than in strain IFO 13350 . Streptomyces griseus HH1 harboring afsA showed a similar level of enzyme activity , however, all the three protease activities sharply increased and the thiol protease activity was critically increased at the end of the fermentation. When a serine protease inhibitor, pefabloc SC, and metalloprotease inhibitor, EDTA, were applied to strain IFO 13350 to examine the in vivo effects of the protease inhibitors on the morpholofical differentiation, the formation of aerial meycelium and spores was delayed by two or three days.

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Effect of Cultivars and Cooking Methods on the Trypsin Inhibitor Activities of Potatoes

  • Kim, Mi-Yeon;Son, Chan-Wok;Shim, Hyun-Jung;Lee, Jeung-Hee;Lee, Kun-Jong;Sok, Dai-Eun;Kim, Hyoung-Chin;Yoon, Won-Kee;Kim, Hwan-Mook;Kim, Mee-Ree
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.161-165
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    • 2008
  • The trypsin inhibitor activities (TIA) of various potato cultivars were evaluated by measuring the inhibition of trypsin inhibitor activity using N-benzoyl-DL-arginine-p-nitroanilide (BAPNA) as substrate. The TIA values of 5 potato cultivars (1.99 to 2.88 mg/g) were significantly different among cultivars (p<0.05). When the TIA values of commercially processed potatoes were determined, no TIA was detected. During cooking, the $IT_{50}$ (time required to reach 50% inhibition of TIA) values were decreased as heating temperature and time increased. The ITso of moist heating was estimated to be 0.34 min at $100^{\circ}C$, whereas for deep-fat frying the $IT_{50}$ was 0.13 min at $180^{\circ}C$ and 5.28 min for oven baking at $100^{\circ}C$. The $IT_{50}$ value of microwave cooking was 0.194 min at medium heat, and which was similar to that of pressure cooking at $120^{\circ}C$ (0.185 min). Moreover, there was a negative relationship between temperature (${\geq}80^{\circ}C$) and $IT_{50}$ values ($R^2=0.99$, p<0.01). The TIA of potato was completely inactivated by moist heating at $100^{\circ}C$ within 5 min, whereas the pressure cooking at $120^{\circ}C$ and deep-fat frying at $180^{\circ}C$ within 60 and 30 sec, respectively. Based on our results, deep-fat frying is the most effective cooking method to reduce TIA in potatoes.

Effects of Heat Treatment on Soybeans With and Without the Gene Expression for the Kunitz Trypsin Inhibitor: Chick Growth Assays

  • Burnham, L.L.;Kim, I.H.;Hancock, J.D.;Lewis, A.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.12
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    • pp.1750-1757
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    • 2000
  • A total of 864 broiler chicks were used at Kansas State University and the University of Nebraska to determine the effects of heat treatment of two soybean genotypes on the growth performance. The soybeans were Williams 82 variety with (+K) and without (-K) gene expression for the Kunitz trypsin inhibitor. Heat treatment (autoclaving at $121^{\circ}C$ and $1.1kg/cm^2$) was applied for 0, 3, 6, 12, 18, and 24 min, resulting in a $2{\times}6$ factorial arrangement of treatments. Station and station treatment effects occurred, indicating that response in nutritional value of the soybean genotypes to heat treatment varied from year to year and location to location. However, the interactions were in magnitude of response rather than direction of response, with greater reductions in trypsin inhibitor concentrations for the soybeans heat processed at the Nebraska location. Pooled data indicated that -K supported greater (p<0.001) ADG, ADFI and gain/feed than the +K genotype. As the length of heat treatment increased, the ADG, ADFI, and the gain/feed ratio increased for chicks fed both soybean genotypes (p<0.0001). However, heating the -K soybeans resulted in a greater response in ADG, ADFI, and gain/feed than heating the +K soybeans (genotype heat treatment interaction, p<0.001). Pancreatic weights (mg pancreas/g of BW) of chicks fed -K soybeans were reduced compared to those from chicks fed +K (p<0.001). Increasing heat treatment decreased pancreas weights in chicks fed both soybean genotypes (p<0.001). Chicks fed heated soybeans in the Nebraska experiment had lower pancreatic weights than chicks fed heated soybeans in the Kansas experiment (station heat treatment interaction, p<0.0001). Chick growth performance was improved and pancreatic weights decreased by feeding raw -K soybeans versus raw +K soybeans, and by increasing heat treatment of both soybean genotypes. However, the response to heat treatment was not independent of genotype. Both +K and -K soybeans heated for 24 min supported similar ADG, ADFI, gain/feed, and pancreas weights, although chicks fed raw +K soybeans had lower growth performance than chicks fed -K soybeans. In conclusion, raw -K soybeans supported greater growth performance in broiler chicks than raw +K soybeans, although this advantage was lost when both soybean genotypes were heated for 24 min. Heat treatment of +K soybeans supported similar growth performance to heated -K soybeans, even though +K soybeans supported lower rates and efficiencies of gain than -K soybeans when fed raw.

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.

A Stack of Recessive Alleles of Kunitz Trypsin Inhibitor, Lectin, and Stachyose in Soybean (콩에서 쿠니츠트립인히비터, 렉틴 및 스타키오스에 대한 열성 유전자의 집적)

  • Choi, Sang Woo;Chae, Won Gi;Kang, Gyung Young;Chung, Jong Il
    • Journal of Life Science
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    • v.29 no.7
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    • pp.774-778
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    • 2019
  • Soybean [Glycine max (L.) Merr.] is one of the major food sources of protein, oil, carbohydrates, isoflavones, and other nutrients for both humans and animals. However, soybean seeds contain antinutritional factors, such as lectin protein, Kunitz Trypsin Inhibitor (KTI) protein, and stachyose. The objective of this research was to stack recessive alleles for development a triple recessive genotype, titilelers2rs2, with low KTI protein, lectin protein, and stachyose contents. Three parents (Gaechuck#2, PI200508, and 14G20) were used to develop the breeding population. The presence or absence of the lectin and KTI proteins was detected by western blotting. The stachyose content in mature seeds was determined by HPLC. Agronomic traits, such as plant type, plant height, maturity date, lodging, seed quality, and 100-seed weight, were evaluated for the four $F_3$ plant strains. One $F_4$ plant strain with the desired agronomical traits was selected. One new strain with the triple recessive titilelers2rs2 genotype was developed. The plant height of the new strain was 51 cm and the 100-seed weight was 31.0 g. The new strain had a yellow seed coat and yellow hilum. The stachyose content of the new strain was 3.8 g/kg. One strain developed in this research will be used to produce improved yellow soybean cultivars that are free of lectin and KTI proteins and low in stachyose content.

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.