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Effect of Antibody Immobilization Method to Magnetic Micro Beads on its Immunobinding Characteristics (자성 미세입자에의 항체 고정화 방법이 면역결합반응에 미치는 영향)

  • Choi, Hyo Jin;Hwang, Sang Youn;Jang, Dae Ho;Cho, Hyung Min;Kang, Jung Hye;Seong, Gi Hun;Choo, Jae Bum;Lee, Eun Kyu
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.65-72
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    • 2006
  • Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Studies on the Chemical Constituents of the New Zealand Deer Velvet Antler Cervus elaphus var. scoticus-(I)

  • Lee, Nam Kyung;Shin, Hyun Jung;Kim, Wan Seok;Lee, Jong Tae;Park, Chae Kyu
    • Natural Product Sciences
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    • v.20 no.3
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    • pp.160-169
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    • 2014
  • 44 compounds and 9 minerals were isolated from and detected in the New Zealand deer velvet antler Cervus elaphus var. scoticus L$\ddot{o}$nnberg. The chemical structures of (1 - 26) were identified on the basis of the spectroscopic methods and comparisons with literature, respectively. The structures were identified as cholesterol (CS, 6), 7-keto-CS (7), $7{\beta}$-hydroxy-CS (8), and $7{\alpha}$-hydroxy-CS (9), and included 12 steroid $3{\beta}$-O-(palmitic/stearic/myristic acid esters; PM/SA/MS) [CS-$3{\beta}$-O-PM (1 - 1), CS-$3{\beta}$-O-SA (1 - 2), CS-$3{\beta}$-O-MR (1 - 3), 7-keto-CS-$3{\beta}$-O-PM (2 - 1), 7-keto-CS-$3{\beta}$-O-SA (2 - 2), 7-keto-CS-$3{\beta}$-O-MR (2 - 3), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-SA (3 -1), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-PM (3 - 2), $7{\beta}$-hydroxy-CS-$3{\beta}$-O-MR (3 - 3), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-SA (4 - 1), $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-PM (4 - 2), and $7{\alpha}$-hydroxy-CS-$3{\beta}$-O-MR (4 - 3)], dinonyl phthalate (5), 8 nucleic acids analogues [uracil (10), deoxyguanosine (11), deoxyuridine (12), uridine (13), deoxyadenosine (14), adenosine (15), inosine (16), and guanosine (17)], and the 9 free amino acids [L-phenylalanine (18), L-isoleucine (19), L-leucine (20), L-tyrosine (21), L-valine (22), L-proline (23), L-threonine (24), L-alanine (25), and L-hydroxyproline (26)]. Also, there are 8 kinds of amino acids [asparagine, serine, glutamine, glycine, histidine, arginine, methionine, and lysine], 2 sialic acids [N-acetylneuraminic acid (27), ketodeoxynonulosonic acid (28)], and 9 minerals [Na > K > Ca > Mg > Fe > Zn > B > Al > Cu] were detected from the autoaminoacid analyzer and ICP spectrometer, HPAEC-PAD/HPLC-FLD, respectively. 9 kinds of oxycholesterol-$3{\beta}$-O-fatty acid ester (2 - 1, 2 - 2, 2 - 3, 3 - 1, 3 - 2, 3 - 3, 4 - 1, 4 - 2, and 4 - 3) and 3 nucleic acids (12, 14, and 15) were isolated from the velvet antler for the first time. 6 kinds of steroids (7, 8, 9, 2 - 1, 3 - 1, and 4 - 1) were examined for their anti-proliferative effects against L1210, P388D1, K562, MEG-01, KG-1, MOLT-4, A549, HepG2, MCF-7, SK-OV-3, and SW-620 cancer cell lines. They showed anti-proliferative effects with $IC_{50}$ values of 0.06, 2.16, 2.42, > 50.0, 1.66 and $8.31{\mu}M$ against L1210, while the values were 24.05, 9.44, 5.22, 0.25. 9.48 and $49.77{\mu}M$ against P388D1, respectively. The others were inactive.

Physiological Interactions Between the Herbicide Pretilachlor and the Safener Fenclorim on Rice (제초제(除草劑) Pretilachlor와 해독제(害毒劑) Fenclorim의 수도(水稻)에 대한 생리적(生理的) 상호작용(相互作用))

  • Han, S.S.;Hatzios, K.K.
    • Korean Journal of Weed Science
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    • v.10 no.4
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    • pp.328-337
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    • 1990
  • The incividual and combined effects of the chloroacetanilide herbicide pretilachlor and of the safener fenclorim on the growth and selected physiological processes of rice (Oryza sativa L., var 'Lemont')were evaluated under greenhouse and laboratory conditions. Fenclorim applied at rates ranging from 50 to 300 g a.i./ha antagonized the injurious effects caused by 150 to 900 g a.i./ha of pretilachlor on 15-day old wet-sown rice grown under greenhouse conditions. When used rates of 150 g/ha or higher, fenclorim reversed completely the effects of all doses of pretilachlor on rice. When the two compounds were given simultaneously, fenclorim enhanced the uptake of $^{14}C$pretilachlor into rice leaf mesophyll protoplasts measured for 1 hr, indicating that competition for uptake at the protoplast level is not involved in the protective action of this safener. The safener-induced stimulation of pretilachlor uptake was particularly evident when fenclorim was used at concentrations of 10, 20 and $40{\mu}M$. Following 4 hr of incubation, individual treatments with pretilachlor inhibited the in vitro incorporation of radiolabeled precursors into proteins, DNA, and lipids of rice leaf protoplasts only when used at the high concentration of $100{\mu}M$M. Individual treatments with high concentrations (10 or $100{\mu}M$) of the safener fenclorim inhibited the incorporation of radiolabeled precursors into proteins and lipids of rice protoplasts, but had no DNA synthesis. The combined effects of pretilachlor and fenclorim on the incorporation of radiolabeled precursors into these macromolecules of isolated rice mesophyll protoplasts appeared to be additive or slightly synergistic rather than antagonistic. Fenclorim at $1{\mu}M$ antagonized the effects of pretilachlor on total lipids of rice leaf protoplasts. In addition, individual and combined treat-menu with pretilachlor and fenclorim influenced the incoroporation of$^{14}C$acetate into polar lipids, triglycerides and steryl esters of rice leaf protoplas causing a redistribution of carbon in these lipid fractions. However, these effects were not large enough to explain the herbicidal activity of pretilachlor or to account for the protective action of the safener fenclorim. Overall, the uesults of the present study idnicate that the safener fenclorim does not seem to protect rice against pretilachlor injury by antagonizing its effects on protein, DNA, or lipid syntheses.

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Comparison of Volatile Components in Organs of Ocimum basilicum L. cultivated in Korea (국내 재배 바질의 품종간 부위별 휘발성 성분 비교)

  • Ahn, Dai-Jin;Lee, Jae-Gon;Kim, Mi-Ju;Lee, Jong-Chul
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.130-138
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    • 2001
  • This study was conducted to compare volatile components from the ten kinds of basils cultivated in Korea. The ten kinds of basils were separated flower, leaf, and stem part from whole plants, respectively. All sample separated were extracted by simultaneous steam distillation-extraction method(SDE) and were analyzed by gas chromatography(GC) and mass selective detector(MSD). Total 42 components were identified in essential oils including 11 alcohols, 6 carbonyls, 20 hydrocarbons and 5 esters components. The major components were linalool, methyl chavicol, eugenol, trans-methyl cinnamate, ${\beta}-cubebene$ and 1,8-cineole. The content of linalool was high significantly in the flower$(31.8{\sim}53.0%)$, the leaf and stem showed $21.8{\sim}35.8%$ and $3.5{\sim}22.4%$, respectively. Especially, the content of methyl chavicol was high relatively in the leaf$(0.4{\sim}32.9%)$, the flower and stem showed $0.2{\sim}24.1%$ and $0{\sim}2.2%)$, respectively. Articock, figz, glove, and greek basils were rich in eugenol$(18.8{\sim}48.7%)$ and poor in methyl chavicol$(0{\sim}5.4%)$ when compared with others kinds of basils. The composition of the components identified showed quite difference between kinds of basils, and the number of components identified in stem was much less than that in flower and leaf

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Study on the Analytical Method and Monitoring of the Oxidized Polyethylene Wax in Foods (식품 중 oxidized polyethylene wax 분석법 연구 및 함유량 실태 조사)

  • Choi, Seung-Hyun;Kim, Jae-Min;Choi, Sun-il;Jung, Tae-Dong;Cho, Bong-Yeon;Lee, Jin-Ha;Lee, Gunyoung;Lim, Ho Soo;Yun, Sang Soon;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.32 no.4
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    • pp.284-289
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    • 2017
  • Oxidized polyethylene wax (OPEW) is, one of the food additives, used as a coating agent in citrus fruits and nuts. OPEW is authorized to quantum satis in EU, USA, and is acceptable less than 250 mg/kg in Australia and New Zealand. But OPEW is unauthorized as a food additive in Korea. This study was to establish the analytical method of OPEW and demonstrate the effective application of various food samples. We first conducted to compare the various analytical method including acid value (AV), high temperature gel permeation chromatography (HT-GPC), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS), gas chromatography flame ionization detector (GC-FID) and fourier transform infrared spectroscopy (FT-IR). This result indicated that FT-IR spectrum of OPEW treated food sample displayed absorption bands for carbonyl group (C=O, $1714cm^{-1}$), ester group (C-O, $1463cm^{-1}$), aliphatic group (C-H, $2916cm^{-1}$). Furthermore, IR spectrum of OPEW treated food sample showed similar tendency with IR spectrum of OPEW standard. Therefore, it is confirmed that analytical method using FT-IR can be detected on analysis of OPEW in food. As a result of monitoring of 111 samples using established analytical method, OPEW was not detected in the food samples.

Quality Characteristics of the Mashes of Takju Prepared Using Different Yeasts (효모종류를 달리한 탁주 술덧의 품질특성)

  • Lee, Heung-Sook;Park, Chang-Sook;Choi, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.56-62
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    • 2010
  • Quality characteristics such as alcohol, acid, and sugar contents of takju brewing mashes prepared using several yeasts were investigated during 12-days of fermentation. Among the yeasts examined, S. cerevisiae led to the highest level of ethanol (10.2-13.4%) and total minor alcohols (0.729-0.831 mg/mL). Regardless of the yeasts used, the acidity showed drastic changes (pH from 3.4-4.2 down to 2.4-2.7) during the first 2 days, and displayed negligible changes from day 4. The total acid contents rapidly increased to the first 4 days of fermentation and leveled off from 4 to the end of fermentation period. The common organic acid components were lactic, succinic and acetic acids. The total and reducing sugar contents varied depending on the yeasts used, with mashes prepared using S. coreanus and S. rouxii producing the maximum total sugar contents (5.43-5.5%) at the end of fermentation. The reducing sugar showed its maximum (7.53-14.89%) at day 2, after which it decreased to its minimum levels (3.04-4.52%). The common free sugar components were glucose and fructose, while S. ellipsoideus led to a higher free sugar level (0.35-5.29%).

Amount and Chemical Characteristics of the Epicuticular Waxes on Leaves at Active Tillering and Heading Stages of Rice Varieties (벼 품종들의 분얼성기 및 출수기의 엽표면Wax의 양 및 화학적 조성)

  • Yong Woong, Kwon;Bong Jin, Chung
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.2
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    • pp.185-197
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    • 1992
  • Differences in the amount and chemical characteristics of the epicuticular waxes on rice leaves were studied for the active tillering and heading stages of rice varieties differing widely in gross leaf-surface property and genetics. The amount of waxes on surfaces of rice leaf-blades was determined by extraction with chloroform and chemical composition of the waxes was characterized by thin layer chromatography, gas liquid chromatography and infrared spectrophotometry. The amount of waxes varied by variety and significantly with growth stage. The amount at the heading stage was 1.7 to 3.6 mg/g fresh weight of leaves, which was two to three times as much as that at the tillering stage of 0.8 to 1.8 mg/g fresh weight. The waxes consisted of seven chemical classes, namely diols, fatty acids, fatty alcohols, fatty aldehydes, fatty esters, saturated and unsaturated hydrocarbons. Diols and unsaturated hydrocarbons were identified as new chemical classes of the rice epicuticular waxes. The polar constituents such as dials, fatty acids and fatty alcohols and the non-polars such as fatty aldehydes, fatty esters, and saturated and unsaturated hydrocarbons were identified at the heading stage, but at the tillering stage only the non-polar compounds were identified. In the carbon numbers (C) of the chemical classes, diols were composed entirely of C30 and acids were mainly of C30 and C31. In alcohols, primary alcohols were composed of C13 and C32, and the secondary alcohols were of C14, C16 and / or C30 regardless of the rice varieties. The acid portion of fatty esters, mainly composed of C22 and C23, showed low cabon numbers compared with the aldehydes. The alcohol portion of them showed a wide distribution in carbon numbers from C13 to C26 depending on the rice varieties. Hydrocarbons had odd carbon numbers, consisting mainly of C29 and C31.

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The effect of air quality and humidity on aged characteristics of beeswax-treated paper during artificial aging (인공열화 시 공기질 및 습도가 밀랍지의 열화에 미치는 영향)

  • Yang, Eun Jeong;Choi, Kyoung Hwa;Kang, Yeong Seok;Cho, Jung hye;Jeong, Hye Young
    • 보존과학연구
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    • s.33
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    • pp.45-55
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    • 2012
  • A beeswax-treated paper has no air permeability but has the water repellency compared with a general Hanji. Because of these properties, the differences of the aging factors and mechanisms between the outer partition of beeswax-treated paper that is affected by the surrounding conservation environment and the inner partition of it that is not affected are bigger than general books. In this research, we analyzed and compared the aging characteristics through the accelerated aging of the beeswax-treated paper by some air and humidity conditions. The results of the physical and optical analysis after the artificial aging, it was shown that the oxygen accelerates the aging of the beeswax-treated paper and the condition with the humidity 50% RH is more stable than the condition with the humidity 0% RH. The results of the CG/MS analysis that was conducted to figure out the decomposition charateristics of the beeswax according to the air quality and the humidity, a low molecular weight compound that the number of carbon is C9-C20 including a fatty acid such as a palmitic acid was increased as the aging was progressed. However, under the same environment, a compound that the number of carbon is C21-C36 including a hydrocarbon and a aliphatic alcohol and a high molecular weight compound that the number of carbon is more than C34 including a wax ester were decreased. A rate of change according to the air quality and the humidity was similar to the beeswax-treated papers.

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Quality characteristics of fermented soybean products produced by lactic acid bacteria isolated from traditional soybean paste (전통 장류 유래 유산균을 이용한 콩 발효물의 품질특성)

  • Lee, Sun Young;Seo, Bo Young;Eom, Jeong Seon;Choi, Hye Sun
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.187-195
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    • 2017
  • This study evaluated quality characteristics of soybean fermented by selected lactic acid bacteria, which were the enzyme strains with high antimicrobial activities isolated from traditionally prepared soybean paste. We determined total aerobic and lactic acid bacteria counts, protease and amylase activities, reducing sugar and amino-type nitrogen contents, and the amounts of amino acids, organic acids, and aroma-compounds. The total aerobic bacteria counts in soybean fermented with strain I13 ($7.75{\times}10^9CFU/mL$) were the highest among all the strains analyzed. Lactic acid bacteria numbers were $2.85{\times}10^9$ to $4.35{\times}10^9CFU/mL$ in soybean fermented with isolates. Amylase and protease activities of the JSB22 sample were the highest among all sample. Reducing sugar and amino-type nitrogen contents of soybean fermented with JSB22 (1.23%, 94.52 mg%) were highest. Total amino acid content of the samples was 15.88-17.62%, and glutamic acid, aspartic acid, leucine, lysine, and arginine were the major amino acids. Lactic acid (0.82-3.65 g/100 g), oxalic acid (22.74-63.57 mg/100 g), and fumaric acid (2.88-6.33 mg/100 g) were predominant organic acids. A total of 39 volatile aroma-compounds were identified, including 2 esters, 5 ketones, 7 alcohols, 14 hydrocarbons, 2 heterocyclic compounds, 4 acids, and 5 miscellaneous compounds. These results represent useful information for the development of a starter (single or complex) and will be used for production of functional fermented soybean foods.