• Korean Journal of Breeding Science
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• v.40 no.2
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• pp.97-100
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• 2008

Lectin protein and Kunitz trypsin inhibitor (KTI) protein of mature soybean seed are a main antinutritional factor in soybean seed. The Le gene controls a lectin protein and Ti gene controls the KTI protein in soybean. Ti locus has been located on linkage group 9 in the classical linkage map of soybean. Position of Le locus on linkage map was not identified. Genetic relationship between Ti locus and Le locus could be useful in soybean breeding program for the genetic elimination of these factors. The objective of this study was to determine the independent inheritance or linkage between Ti locus and Le locus in soybean seed. Two $F_2$ populations were developed from three parents (Gaechuck#1, T102, and PI548415). The $F_1$ seeds from Gaechuck#1 (titiLeLe) ${\times}$ T102 (TiTilele) and Gaechuck#1 (titiLeLe) ${\times}$ PI548415 (TiTilele) were obtained. The lectin and KTI protein were analysed from $F_2$ seeds harvested from the $F_1$ plants to find independent assortment or linkage between Ti locus and Le locus. The segregation ratios of 3 : 1 for Le locus (129 Le_ : 44 lele) and Ti locus (132 Ti_ : 41 titi) and were observed. The segregation ratios of 9 : 3 : 3 : 1 (95 Le_Li_ : 34 Le_titi: 37 leleTi_ : 7 leletiti) between Le gene and Ti gene in $F_2$ seeds were observed. This data showed that Ti gene was inherited independently with the Le gene in soybean. These results will be helpful in breeding program for selecting the line with lacking both KTI and lectin protein in soybean.

• Korean Journal of Food Science and Technology
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• v.15 no.2
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• pp.101-107
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• 1983

The four varieties of Korean soybeans were allowed to undergo natural fermentation for seven days at ambient temperature. The average pH of the product was 3.93 and titratable acidity was 1.94%. For all varieties of soybeans the content of riboflavin increased from 98 to $309.4{\mu}g/100g$ dry-matter, relative nutritive value from 78.66 to 94.59% and available lysine from 6.56 to 7.38 mg/gN, respectively. During fermentation, the activities of protease and lipase increased, while lipoxygenase and trypsin inhibitor activity decreased markedly. The capacity of water sorption of fermented soybean flour was increased with progress of proteolysis during fermentation. The cookie and noodle prepared with 20:80 mixture of fermented soybean flour and wheat flour were in the 'like' category, but it was desirable to neutralize the sour taste produced by fermentation. Among five kinds of products prepared from the fermented soybean flour pan cake was liked most by rural consumers.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.3
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• pp.299-306
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• 2009

In nitrogen-limited conditions, rhizobia lead to formation of nitrogen-fixing nodules on the roots of leguminous plants. The process of nodulation is autoregulated by pre-existing nodules in the same root system. The altered profile of sap proteins by inoculation with B. japonicum may indicate presence of a signal responsible for autoregulation transferred through stem. The 20 kDa protein enhanced by innoculation significantly decreased in intensity from 2.5 to 7 days after inoculation (DAI). However 6 kDa protein did increase during such a transition period. Western blot analysis showed that both 20 kDa and 6 kDa were cross-reacted with the SKTI antiserum. This suggests that SKTI may be involved in soybean nodulation by specific induction and degradation in stem sap during early stage of nodulation. RNAi technique and Agrobacterium rhizogenes-mediated transformation were applied to investigate the function of SKTI in nodulation. We have found that the number of rhizobium-induced nodule was much less in SKTIi-silenced hairy roots than the non-silenced. Indeed the quantitative RT-PCR showed that the expression level of SKTI gene was reduced over 40% in the transgenic hairy roots compared to the non-transgenic. It appears that the observed early induction of SKTI and degradation into small peptide in a specific time manner may be involved in autoregulation of nodulation in soybean and the specific mechanism of such regulation remains to be investigated.

• BMB Reports
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• v.28 no.2
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• pp.138-142
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• 1995

An anticoagulant/fibrinolytic protease was purified to homogeneity from the earthworm Lumbricus rubellus. The protein was a single chain glycoprotein of 32 kDa that exhibited strong proteolytic activity on human thrombin and fibrin clots. Proteolytic degradation of these plasma proteins by the purified enzyme occurred at a neutral pH range. Among several human plasma proteins tested as possible substrates for the protease reaction, the 32 kDa enzyme specifically hydrolyzed both thrombin and fibrin polymers without affecting other proteins, such as serum albumin, immunoglobulin, and hemoglobin. Treatment of the purified enzyme at neutral pH with either phenylmethylsulfonylfluoride or soybean trypsin inhibitor resulted in a loss of catalytic activity. The enzyme hydrolyzed the chromogenic substrate H-D-Phe-L-Pipecolyl-L-Arg-p-nitroanilide with a $K_m$ value of 1.1 ${\mu}M$ at a neutral pH. These results suggest that the anticoagulant/fibrinolytic enzyme from Lumbricus rubellus is a member of the serine protease family having a trypsin-like active site, and one of the potential clevage sites for the enzyme is the carbonyl side of arginine residues in polypeptide chains.

• Journal of Life Science
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• v.22 no.4
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• pp.524-531
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• 2012

Soybean is one of the most common food materials causing food hypersensitivity reactions in Korea. In this study, we have investigated the effect of roasting and fermentation on the allergenicity of soybean. Three kinds of soybean ($Daepung$, $Daewon$, and $Taegwang$) were prepared as raw, roasted, and fermented by $Bacillus$ $subtilis$ GSK 3580, and then their proteins were extracted. The proteins were separated using SDS-PAGE, and the detection of IgE specific to soybean proteins was performed by immunoblotting using 7 sera of soybean allergy patients and non-allergic control individuals. Serum specific IgE to soybean was measured by ELISA. The SDS-PAGE of raw soybean proteins showed various-sized bands ranging from 9 to 76 kDa, which are known as major allergens. In particular, 9, 21, 34, 52, 72, and 76 kDa proteins are known as LTP, Kunits trypsin inhibitor, $Gly$ m Bd 30K, ${\beta}$-subunit, ${\alpha}$-subunit, and ${\alpha}$'-subunit of ${\beta}$-conglycinin, respectively; these are major allergens in soybean. In contrast, only peptides of less than 35 kDa were found in roasted and fermented soybeans. IgE immunoblot analysis of three roasted species of soybeans commonly detected at 38-40 kDa and 10-15 kDa. The protein bands in fermented soybean showed very weak signals or were not detected. In addition, the reactivity of most patients' sera to soybean was decreased after roasting and fermentation. With these results, it may be concluded that the allergenicity of soybeans is reduced by the roasting and fermentation processes. It is supposed that allergenic proteins in soybean were degraded by heat treatment methods and proteolytic enzymes were secreted from fermenting microorganisms.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.279-279
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• 1994

Fibrinolytic proteases, piscivorase I (PI) and piscivorase II (PII), were isolated from Agkistrodon piscivorus piscivorus (eastern cotonmouth moccasin) venom using gel filtration on Bio-Gel P100 and ion-exchange chromatography on CM-Sepharose. The molecular welghts of two proteases were approximately 23400 and 29000. Their isoelectric points 6.6 and 8.5, respectively. The partial amino acid sequences of PI were characterized by tryptic digestion. PI readily cleaves the A${\alpha}$-and B${\beta}$-chaln of fibronogen, but PII rapidly cleaves A${\alpha}$-chain and more slowly the B${\beta}$-chain, They were activated by Ca$\^$2+/, Mg$\^$2+/ and Ba$\^$2+/, but inhibited by Zn$\^$2+/, Cu$\^$2+/ and Mn$\^$2+/. Two enzymes were also inhibited by cysten, ${\beta}$-mercapto -ethanol, and by metal chelators such as EDTA and EGTA, but not by benzamidine, PMSF, soybean trypsin inhibitor and aprotinin. They did not act like thrombin, plasmin and kallikrein, using specific chromogenllc substrates. Two protease did not induce platelet aggregation. PI showed low hemorrhagic activity at dosage of 50 $\mu\textrm{g}$.

• Mycobiology
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• v.31 no.4
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• pp.209-213
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• 2003

Aspergillus flavus K-03 isolated from poultry forming soil in Korea was studied for its ability to produce extracellular proteases on basal medium containing 2%(w/v) chicken feathers. The fungus was observed to be a potent producer of such enzymes. Keratinolytic enzyme secretion was the best at 15 days of incubation period at pH 9 and temperature $40^{\circ}C$. No relationship existed between the enzyme yield and increase of biomass. Enzyme production was suppressed by exogenous sugars in descending order arabinose>maltose>mannose>fructose. But glucose did not influence the enzyme activity. The keratinolytic enzyme released by the fungus demonstrated the ability to decompose keratin substrates as chicken feather when exogenous glucose was present. The keratinolytic activity was inhibited by $HgCl_2$ and serine-protease inhibitors such as phenymethylsulfonyl fluoride(100%), chymostain(88%), crystalline soybean trypsin inhibtor(80%), antipain(45%) and aprotinin(40%), and was not by cystein-protease and aspartyl-protease inhibitors. The enzyme activity is only partially inhibited by metallo-protease inhibitor. Thus, the enzyme secreted by A. flavus K-03 belongs to the alkaline serine-type protease.

• BMB Reports
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• v.35 no.2
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• pp.165-171
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• 2002

A serine collagenolytic protease was purified from the internal organs of filefish Novoden modestrus, by ammonium sulfate, ion-exchange chromatography on a DEAE-Sephadex A-50, ion-exchange rechromatography on a DEAE-Sephadex A-50, and gel filtration on a Sephadex G-150 column. The molecular mass of the filefish serine collagenase was estimated to be 27.0 kDa by gel filtration and SDS-PAGE. The purified collagenase was optimally active at pH 7.0-8.0 and $55^{\circ}C$. The purified enzyme was rich in Ala, Ser, Leu, and Ile, but poor in Trp, Pro, Tyr, and Met. In addition, the purified collagenolytic enzyme was strongly inhibited by N-P-toluenesulfonyl-L-lysine chloromethyl ketone (TLCK), diisopropylfluorophosphate (DFP), and soybean trypsin inhibitor.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1598-1604
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• 2001

Eight $12.4{\pm}0.6kg$ initial body weight crossbred barrows were used to determine the effect of soybean galactooligosaccharides on nutrient and energy digestibility, and digesta transit time. Four dietary treatments were utilized in this trial. Treatment one was a corn-soybean meal based diet (SBM) containing raffinose and stachyose at the levels of 0.16% and 0.75%, respectively. Treatment two (control) was a corn-HP300 (soybean concentrate protein) diet. In treatments three and four, 1.1% and 2.2% commercial stachyose was added to the control diet to provide total dietary stachyose at the levels of 1% and 2%, respectively. The soybean galactooligosaccharides (raffinose + stachyose) level in treatment one was slightly lower compared to that in treatment three. Three collection periods were run with two pigs for each treatment/period. There was a 4 d adjustment period followed by a 3 d collection period. The results showed that the nitrogen retention (86.79%) of pigs fed treatment two diet was higher than that of pigs fed treatment one by 5.2% (p<0.05). The nitrogen retention of treatment three was intermediate 83.09%. The apparent fecal digestibility of all amino acids in treatment two was numerically highest, followed by treatments three and four. However, there were no significant difference among groups (p>0.05). The dry matter (DM), organic matter (OM), crude protein (CP), and crude fiber (CF) digestibility numerically decreased as the soybean galactooligosaccharides level increased, but were not significantly different (p>0.05). Chromium content in feces (from the inclusion of 0.3% chromic oxide in the diets) differed among treatments (p<0.05) at 15 h, 18 h, and 21 h after eating. This showed that the digesta transit time was differed significantly among treatments. Treatment four was the shortest, followed by treatment three, SBM and control. The results demonstrated that in the absence of antinutritional factors and soybean antigen protein, inclusion of 1% and 2% stachyose in corn-HP300 diet has no significant effect on the digestibility of DM, OM, CP, CF and amino acids. When the soybean galactooligosaccharide level in diet one and diet three were adjusted to be almost the same, antinutritional factors such as trypsin inhibitor and soybean antigen protein could decrease the nutrient digestibility and nitrogen retention rate of diet. High levels of soybean galactooligosaccharides shortened the digesta transit time in the intestinal tract. This trial suggested that the total level of soybean galactooligosaccharides (stachyose+raffinose) in the weanling piglet diet is better not to exceed 1% when common soybean meal is used as main protein source.

• Journal of the Korean Society of Food Science and Nutrition
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• v.11 no.4
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• pp.7-12
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• 1982

Growth of soybean sprouts(Glycine Max L.) and amounts of some chemical components were measured when they were exposed to blue light (120lux, 3hrs/day) during their growth. Hypocotyl length of irradiated soybean sprouts exceeded slightly that of control (dark) soybean sprouts, but the tfresh weight of whole sprouts as well as each part of the sprout showed no difference between the two groups. Chlorophyll content of cotyledon under blue light increased significantly with the lapse of days (3.57 and $8.45\;{\mu}g/100g$ fresh weight on the 3rd and 7th day). Bluelight irradiated sprouts contained more vitamin C than control sprouts (21.7% and 30.8% higher for the cotyledon and hypocotyl). Total amount of protein was not affected. Hypocotyl protein content was 8% of that in original soybean. Blue light did not affect the activity of trypsin inhibitor of sprouts. Similar activity of the inhibitor was observed in the cotyledon whereas hypocotyl showed activity corresponding to 23.7% of original bean. Polyacrylamide gel electrophoretogram for the protein showed 10, 9, and 11 bands in the original bean. 5th day cotyledon and hypocotyl respectively. Especially, band 3 of low Rm value was major protein component for the hypocotyl. Band 5 and 11 could be seen only in the protein of hypocotyl from bluelight irradiated sprouts, whereas no effect of blue light on the electrophoretogram was observed for the cotyledon.