• BMB Reports
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• v.40 no.3
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• pp.373-381
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• 2007

Seed storage proteins (SSPs) are important for seed germination and early seedling growth. We studied the accumulation and degradation profiles of four major Arabidopsis 12S SSPs using a 2-DE scheme combined with mass spectrometric methods. On the 2-DE map of 23 dpa (days post anthesis) siliques, 48 protein spots were identified as putative full-length or partial $\alpha$, $\delta$ subunits. Only 9 of them were found in 12 dpa siliques with none in younger than 8 dpa siliques, indicating that the accumulation of 12S SSPs started after the completion of cell elongation processes both in siliques and in developing seeds. The length and strength of transcription activity for each gene determined the final contents of respective SSP. At the beginning of imbibition, 68 SSP spots were identified while only 2 spots were found at the end of the 4 d germination period, with $\alpha$, subunits degraded more rapidly than the $\alpha$ subunits. The CRC $\delta$ subunit was found to degrade from its C-terminus with conserved sequence motifs. Our data provide an important basis for understanding the nutritional value of developing plant seeds and may serve as a useful platform for other species.

• Food Science and Biotechnology
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• v.17 no.1
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• pp.135-143
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• 2008

The ratio between 11S Sand 7S globulins was greater in large seed size varieties (LSSVs) of soybean than in small seed size varieties (SSSVs) or medium seed size varieties (MSSVs) but did not differ between MSSVs and SSSVs. The cysteine and methionine contents of 11S globulins were greater than those of total seed proteins and 7S globulins. The acidic monoamino monocarboxylic amino acids were the most abundant class of amino acid in soybean seed (27.2%) and soybean curd protein (29.3%). Isolated 7S and 11S fractions were analyzed by HPLC. Of the 12 peaks detected, 4 constituted 64.1% of the proteins of the SSSVs, 65.6% of the proteins of the MSSVs, and 70.5% of the proteins of the LSSVs. The 11S/7S globulin ratio was related to the yield and hardness of soybean curd. The MSSVs had the greatest yield of soybean curd, but the soybean curd hardness of the MSSVs was greater than that of the SSSVs. These results show that the 11S/7S ratio and color of soybean seeds can be used to predict the yield, hardness, and color of soybean curd.

• Proceedings of the Botanical Society of Korea Conference
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• 1987.07a
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• pp.283-307
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• 1987

Glycinin and $\beta$-conglycinin are the most abundant storage protein in soybean. These proteins are known to be synthesized predominantly during germination and cell expansion phase of seed development for short period, and synthesized not in other tissues. Genes encoding these storage proteins are useful system to study the mechanism of development stage and tissue specific gene expression in eukaryotes, especially plants, at the molecular level. The cDNA and genomic clones coding for glycinin have been isolated and regulation mechanism of the gene expression has been studied. Initially, development and tissue-specific expression of the glycinin gene is regulated at the level of transcription. Post-transcriptional processing is also responsible for delayed accumulation of the mRNA. Translational control of the storage protein gene has not been reported. Post-translational modification is another strategic point to regulate the expression of the gene. It is possible to identify positive and/or negative reguratory clements in vivo by producing transgenic plants agter gene manipulation. Elucidation of activation and repression mechanism of soybean storage protein genes will contribute to the understanding of the other plant and eukaryotic genes at molecular level.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.293-293
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• 2017

The objective of this study was to determine physicochemical changes in brown rice during ageing condition. Five varieties (Haiami, Ilpum, Daecheong, Jungwon, and Dasan1) of brown rice were stored at $35^{\circ}C$ for 8 weeks. Crude protein and lipid content, seed germination rate, fat acidity, tocol content, TOYO glossiness value, pasting properties, and composition of storage proteins were measured to evaluate its quality during storage. The isomers of tocols (tocopherol and tocotrienols) were quantified using HPLC system, and the pattern of variation in rice storage proteins was examined through electrophoresis of protein extracts. Seed germination rate decreased by 2.7 times, whereas the fatty acid value dramatically increased by 4.8 times after 8 weeks of storage. Toyo glossiness value of cooked milled rice considerably affected by storage period, and the pasting properties of milled rice were also influenced by storage. The final viscosity and breakdown value increased, but setback decreased during storage. In terms of storage protein, proportion of prolamin (14.3 kDa) and globulin (26.4 kDa) increased, whereas percentage of glutelin (34-39 kDa and 21-22 kD) decreased. Furthermore, the contents of total tocol and isomers decreased in stored brown rice.

• 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.

• Journal of Plant Biology
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• v.39 no.2
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• pp.123-126
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• 1996

The pattern of seed storage protein, vicilin, deposition and site of intracellular localization was examined in cotyledon cells of pea (Pisum sativum) seed using the immunocytochemical methods. The vicilin was confined to the cisternae fo the rough endoplasmic reticulum and dictyosome as well as protein granules newly formed in rough endoplasmic reticulum. Vacuolar protein deposites and protein bodies were also labelled by gold particles. After small protein bodies were formed in the rough endoplasmic reticulum, they were transported to large protein bodies and then fused together. Electron dense protein granule, elaborated in the dictyosome, appears to be transported from dictyosome to protein body. A few unlabelled protein granules seem to be accumulated in other type of proteins than vicilin.

• Journal of Plant Biology
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• v.37 no.3
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• pp.293-299
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• 1994

Developing rice endosperm cells display two morphologically distinct rough endoplasmic reticulum (ER) membranes, the cisternae ER (C-ER) and theprotein body ER (PB-ER), the latter delimiting the prolamine protein bodies. We (Li et al., 1993) have recently shown that the storage protein mRNAs are not randomly distributed on these ER types; the C-ER is enriched for glutelin mRNAs, whereas the PB-ER harbors predominantly prolamine transcripts. To address whether these ER types have differnet capacities to translate these mRNAs and translocate their proteins into the lumen, a microsomal fraction enriched in C-ER vesicles was prepared from devleoping rice seeds. When present in an in vitro translatin system, the microsomes were able to proteolytically remove the signal peptide and internalize both preproglutelin and preprolamine within the microsomal vesicles. Of the two species, preprolamine was more effectively translocated and processed. These results suggest that the C-ER has the capacity to recognize and bind both storage protein mRNAs during protein synthesis. Moreover, efficient translocation and processing of glutelin requires additional factors that are deficient or absent in the in vitro system.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.92-102
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• 2006

Abundant proteins often cause problems in proteome study. Glutelin family proteins (hereafter referred to glutelin) are present in rice proteome sample as over-whelming constituents with very high abundance. In order to increase the number of identified proteins in rice proteome study, we developed a newly improved method for sample preparation through the removal of glutelin. When the protein samples from rice seed were extracted by the conventional trichloroacetic acid (TCA) extraction method, glutelin accounts for about 60% of total rice seed proteins in SDS gels. Using our new water extraction method, glutelin consists of only about 10% of total proteins. After analyzing on a two-dimensional gel electrophoresis (2-DE), 937 protein spots were detected using the conventional TCA extraction method. On the other hand, 1240 proteins could be seen using the new water extraction method. The selectivity for non-glutelin and less abundant protein by the water extraction method was also confirmed by ESI-Q/TOF mass spectrometry analysis. Thus, the new water extraction method developed here can be efficiently used to study the proteome analysis of rice storage seed.

• Journal of Food Hygiene and Safety
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• v.6 no.1
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• pp.67-72
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• 1991

This study was carried out to investigate the preservative effect of grapefruit seed extract on fish meat product. he effect of GFSE has been tried on fish meat product divided into three lots; Control (no treatment), 500 ppm and 1,000 ppm of GFSE-treated samples. The results were summarized as follows; 1 During the storage of fish meat product, chagnges of crude protein contents of GFSEtreated samples were smaller than the control. 2 Texture was inclined to decrease as the storage period goes, the decrease ratio of GFSEtreated samples was smaller than the control. 3. SDS-PAGE patterns of fish meat product treated with or without GFSE showed that Mw 30,000-32,000 of major proteins were hydrolyzed and disappeared. The deterioration of fish meat product proteins occured in the storage period of 2 days in the control and 4-5 days in GFSE-treated samples. Conclusivery, the excellent preservative effects of GFSE on fish meat product were shown in the rheological, chemical test and sensory evaluation.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.13-19
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• 1999

The visco-elastic properties of gluten are major determinants of the processing properties of doughs. These visco-elastic properties are strongly influenced by the ratio of monomeric and polymeric proteins and the size distribution of the polymeric proteins, which make up the gluten fraction of the dough. Recent studies have revealed that other features, such as the number of the cysteine residues of the HMW-GS, also play an important role in determining the functional characteristics. To modify the processing properties at molecular level, the relationship between the structure of molecules and dough properties has to be understood. In order to explore the relationships between individual proteins and dough properties, we have developed procedures for incorporating bacterially expressed proteins into doughs, and measuring their functional properties in small-scale equipment. A major problem in investigating the structure/function relationships of individual seed storage proteins is to obtain sufficient amounts of pure polypeptides from the complex families of proteins expressed in the endosperm. Therefore, we have established a simplified model system in which we produce specific protein genes through bacterial expression and test their functional properties in smallscale apparatus after incorporation into base flour. An S poor protein gene has been chosen as a template gene. This template gene has been modified using standard recombinant DNA techniques in order to test the effects of varying the number and position of cysteine residues, and the size of the protein. Doughs have been mixed in small scale apparatus and characterized with respect to their polymeric composition and their functional properties, including dough mixing, extensibility and small scale bating. We conclude that dough characteristics can be manipulated in a predictable manner by altering the cysteine residues and the size of high molecular weight glutenins.