• 약학회지
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• 제53권2호
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• pp.69-73
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• 2009

Acute betaine treatment induces time-dependent changes in the hepatic glutathione (GSH), cysteine and S-adenosylmethionine (SAM) levels. Our previous study demonstrated that betaine administered $1{\sim}4$ hours prior to sacrifice decreased hepatic GSH levels, but these levels were increased when measured 24 hours following the treatment. The present study was aimed to determine dose-dependent effects of betaine on hepatic metabolism of sulfur amino acid in mice. Mice were sacrificed 2.5 or 24 hours after intraperitoneal treatment with betaine at different dose levels ranging from 50 to 1000 mg/kg. The concentrations of methionine and SAM were increased by a betaine dose of 100 mg/kg, and the concentrations of GSH and cysteine were decreased by a betaine dose of 200 mg/kg at 2.5 hours. These changes were augmented with increasing doses of betaine. At 24 hours following betaine treatment, increased GSH and decreased taurine levels were observed from dose levels of 400 mg/kg. Changes in hepatic activities of cystathionine beta-synthase, gammaglutamylcysteine ligase and cysteine dioxygenase were observed from dose levels of $200{\sim}400$ mg/kg of betaine administered 24 hours prior to sacrifice.

• Mycobiology
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• 제47권4호
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• pp.441-448
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• 2019

Two new SAM-dependent methyltransferase encoding genes (fvsmt1 and fvsmt2) were identified from the genome of Flammulina velutipes. In order to make a comprehensive characterization of both genes, we performed in silico analysis of both genes and used qRT-PCR to reveal their expression patterns during the development of F. velutipes. There are 4 and 6 exons with total length of 693 and 978 bp in fvsmt2 and fvsmt1, respectively. The deduced proteins, i.e., FVSMT1 and FVSMT2 contained 325 and 230 amino acids with molecular weight 36297 and 24894 Da, respectively. Both proteins contained a SAM-dependent catalytic domain with signature motifs (I, p-I, II, and III) defining the SAM fold. SAM-dependent catalytic domain is located either in the middle or at the N-terminal of FVSMT2 and FVSMT1, respectively. Alignment and phylogenic analysis showed that FVSMT1 is a homolog to a protein-arginine omega-N-methyltransferase, while FVSMT2 is of cinnamoyl CoA O-methyltransferase type and predicted subcellular locations of these proteins are mitochondria and cytoplasm, respectively. qRT-PCR showed that fvsmt1 and fvsmt2 expression was regulated in different developmental stages. The maximum expression levels of fvsmt1 and fvsmt2 were observed in stipe elongation, while no difference was found in mycelium and pileus. These results positively demonstrate that both the methyltransferase encoding genes are involved in the stipe elongation of F. velutipes.

• Biomolecules & Therapeutics
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• 제26권2호
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• pp.167-174
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• 2018

Alterations in sulfur amino acid metabolism are associated with an increased risk of a number of common late-life diseases, which raises the possibility that metabolism of sulfur amino acids may change with age. The present study was conducted to understand the age-related changes in hepatic metabolism of sulfur amino acids in 2-, 6-, 18- and 30-month-old male C57BL/6 mice. For this purpose, metabolite profiling of sulfur amino acids from methionine to taurine or glutathione (GSH) was performed. The levels of sulfur amino acids and their metabolites were not significantly different among 2-, 6- and 18-month-old mice, except for plasma GSH and hepatic homocysteine. Plasma total GSH and hepatic total homocysteine levels were significantly higher in 2-month-old mice than those in the other age groups. In contrast, 30-month-old mice exhibited increased hepatic methionine and cysteine, compared with all other groups, but decreased hepatic S-adenosylmethionine (SAM), S-adenosylhomocysteine and homocysteine, relative to 2-month-old mice. No differences in hepatic reduced GSH, GSH disulfide, or taurine were observed. The hepatic changes in homocysteine and cysteine may be attributed to upregulation of cystathionine ${\beta}-synthase$ and down-regulation of ${\gamma}-glutamylcysteine$ ligase in the aged mice. The elevation of hepatic cysteine levels may be involved in the maintenance of hepatic GSH levels. The opposite changes of methionine and SAM suggest that the regulatory role of SAM in hepatic sulfur amino acid metabolism may be impaired in 30-month-old mice.

• Bulletin of the Korean Chemical Society
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• 제28권3호
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• pp.379-385
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• 2007

Receptor-oriented pharmacophore-based in silico screening is a powerful tool for rapidly screening large number of compounds for interactions with a given protein. Inhibition of the enzyme catechol-Omethyltransferase (COMT) offers a novel possibility for treating Parkinson's disease. Bisubstrate inhibitors of COMT containing the adenine of S-adenosylmethionine (SAM) and a catechol moiety are a new class of potent and selective inhibitor. In the present study, we used receptor-oriented pharmacophore-based in silico screening to examine the interactions between the active site of human COMT and bisubstrate inhibitors. We generated 20 pharmacophore maps, of which 4 maps reproduced the docking model of hCOMT and a bisubstrate inhibitor. Only one of these four, pharmacophore map I, effectively described the common features of a series of bisubstrate inhibitors. Pharmacophore map I consisted of one hydrogen bond acceptor (to Mg2+), three hydrogen bond donors (to Glu199, Glu90, and Gln120), and one hydrophobic feature (an active site region surrounded by several aromatic and hydrophobic residues). This map represented the most essential pharmacophore for explaining interactions between hCOMT and a bisubstrate inhibitor. These results revealed a pharmacophore that should help in the development of new drugs for treating Parkinson's disease.

• Journal of Nutrition and Health
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• 제42권5호
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• pp.423-433
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• 2009

본 연구는 흰쥐를 대상으로 엽산결핍 또는 엽산결핍/비타민 $B_{12}$결핍/0.3% 호모시스틴 식이의 공급을 통해 각각 경미한 호모시스테인혈증과 중위의 호모시스테인혈증을 유도한 후, 혈장, 간 및 뇌 조직내 메티오닌 대사회로 biomarker와 엽산 농도 사이의 상관관계를 분석함으로써 영양성 호모시스테인혈증의 특성을 규명할 목적으로 실시하였다. 6 주령 Sprague-Dawley 숫컷 쥐에게 엽산이 충분한 식이 (FS), 엽산결핍식이 (FD), 또는 동일 식이에 호모시스틴을 첨가한 식이 (FSH and FDH), 엽산결핍/비타민 $B_{12}$ 결핍/호모시스틴 첨가 식이 (FDHCD)를 8주간 공급하였다. 1) FD와 FDH 식이군은 경미한 호모시스테인혈증을 (17.41 ${\pm}$ 1.94 nmol/mL) 나타냈으며, FDHCD 식이군은 중위의 호모시스테인혈증을 (44.13 ${\pm}$ 2.65 nmol/mL) 나타내어 엽산과 비타민 $B_{12}$결핍에 의한 영양성 호모시스테인혈증의 모델로 이용할 수 있었다. 2) FD, FDH, FDHCD 식이군의 간 (p < 0.001)과 뇌조직 (p < 0.01) 내 엽산 농도는 FS, FSH군 보다 유의적으로 낮았으나, FD, FDH, FDHCD 식이군의 간 및 뇌조직의 엽산농도 사이에는 유의적인 차이가 없었다. 이와 대조적으로 혈장 엽산 농도는 FDHCD 식이군 (126.5 ${\pm}$ 9.6 nmol/L)이 FD, FDH 식이군 (21.1 ${\pm}$ 1.4 nmol/L, 22.0 ${\pm}$ 2.2 nmol/L)(p < 0.001) 보다 약 6배 높았으며, 이는 비타민 $B_{12}$ 결핍에 의한"methyl-folate trap"으로 인해 엽산이 효율적으로 조직내 보유되지 못하고 혈류로 나와 소변을 통해 배설되기 때문인 것으로 보인다. 3) FD와 FDH 식이군의 간조직의 SAH 농도는 각각 대조군 보다 44% 및 50%씩 증가되었고 (p < 0.001), 간 SAM 농도는 각각 대조군 보다 72%, 71% 저하되었으며 (p < 0.001), 그 결과 두 군 모두 SAM/SAH 비율이 대조군 보다 약 80% 저하되었다 (p < 0.001). 한편 FDHCD 식 이군의 간 SAH 농도를 대조군과 비교할 때 대조군 보다 107% 증가되었고 (p < 0.001), SAM 농도는 대조군 보다 81% 저하되었으며 (p < 0.001), 그 결과 SAM/SAH 비율이 대조군 보다 약 90% 저하되어 매우 낮은 SAM/SAH 비율을 나타냈다 (p < 0.001). 뇌조직 SAM 농도는 엽산결핍, 비타민 $B_{12}$결핍 및 호모시스틴 급여에 의해 영향을 받지 않고 대조군과 유사한 수준을 보여 뇌조직내 SAM의 항상성을 나타냈으나, FDHCD 식이군의 뇌조직 SAH 농도는 대조군 보다 60% 증가되었으며 (p < 0.05), 그 결과 SAM/SAH 비율은 대조군 보다 약 28% 저하되었다 (p < 0.05). 따라서 중위의 호모시스테인혈증을 나타낸 실험군에서만 뇌조직의 SAH 농도가 증가되었다. 4) 영양결핍 (엽산 또는 비타민 $B_{12}$)에 의한 호모시스테인혈증의 특성을 조사할 목적으로 혈장, 간 및 뇌 조직내 메티오닌 대사회로 biomarker와 엽산 농도 사이의 상관관계를 조사하였다. 혈장 호모시스테인은 간 엽산과 음의 상관관계 (r = -0.641, p < 0.01)을 보였으나, 뇌 엽산 또는 혈장 엽산과는 유의적인 상관관계를 보이지 않았다. 그러나 이와 대조적으로 FDHCD 식이군을 제외시킨 나머지 네개의 실험군 자료만으로 상관관계를 분석하였을 때 혈장 호모시스테인과 뇌 엽산 (r = -0.321, p < 0.05), 혈장 엽산 (r = -0.581, p < 0.01), 간 엽산 (r = -0.684, p < 0.01) 사이에 모두 유의적인 상관관계를 나타냈다. 혈장 호모시스테인과 간조직의 SAH 및 SAM 농도 사이의 상관관계는 FDHCD군을 제외시킨 나머지 네개의 실험군 자료만으로 상관관계를 분석하였을 때 뇌조직 SAH 농도들 제외한 모든 상관계수가 전체실험군 자료로 분석한 경우보다 높았다. 따라서 엽산결핍/비타민 $B_{12}$결핍으로 인한 호모시스테인혈증 (FDHCD)에서는 혈장 엽산이 엽산결핍군 (FD, FDH)보다 높으면서 동시에 혈장 호모시스테인 농도도 높은 특성을 보였다. 결론적으로, 식이 중 엽산만 결핍된 경우와 엽산결핍과 비타민 $B_{12}$ 결핍을 동반할 경우"methyl-folate trap"으로인해 혈장 엽산과 호모시스테인 농도 패턴에 차이가 있었으며, 메티오닌 대사회로의 biomarker 사이의 상관관계와 혈장 엽산, 뇌 엽산 및 뇌 SAH와 호모시스테인 농도 사이의 상관관계가 차이가 있었다. 또한 엽산 결핍과 비타민 $B_{12}$ 결핍으로 인해 나타나는 뇌 SAH 농도의 증가는 메틸화를 저해시킴으로써 인지능력에 영향을 줄 수 있을 것으로 사료된다.

• Bulletin of the Korean Chemical Society
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• 제28권6호
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• pp.941-946
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• 2007

Bacillus halodurans O-methyltransferase (BhOMT) is a S-adenosylmethionine (SAM or AdoMet) dependent methyltransferase. Three dimensional structure of the BhOMT bound to S-adenosyl-L-homocysteine (SAH or AdoHcy) has been determined by comparative homology modeling. BhOMT has 40% sequence identity with caffeoyl-CoA 3-O-methyltransferase (CCoAOMT) from alfalfa. Based on x-ray structure of CCoAOMT, three dimensional structure of BhOMT was determined using MODELLER. The substrate binding sites of these two proteins showed slight differences, but these differences were important to characterize the substrate of BhOMT. Automated docking study showed that four flavonoids, quercetin, fisetin, myricetin, and luteolin which have two hydroxyl groups simultaneously at 3'- and 4'-position in the B-ring and structural rigidity of Cring resulting from the double bond characters between C2 and C3, were well docked as ligands of BhOMT. These flavonoids form stable hydrogen bondings with K211, R170, and hydroxyl group at 3'-position in the Bring has stable electrostatic interaction with Ca2+ ion in BhOMT. This study will be helpful to understand the biochemical function of BhOMT as an O-methyltransferase for flavonoids.

• Nutritional Sciences
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• 제9권2호
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• pp.106-111
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• 2006

Chronic alcohol consumption is associated with perturbation of hepatic metabolism of sulphur-containing amino acid. The goal of present study was to evaluate the influence of dietary supplementation of methionine or folate to chronically ethanol-fed mts on the metabolism of sulfur-containing amino acids and one-carbon metabolism. Sprague-Dawley male mts were fed Lieber-Decarli liquid diet with 0% ethanol (control), 36% ethanol (E), 36% ethanol combined with methionine supplement (EM) or folate supplement (EF) for 8 weeks. Hepatic S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), plasma folate and homocysteine (Hcy), urinary excretion of folate and formiminoglutamate were investigated after feeding experimental diets. Growth was retarded by 36% ethanol consupmtion (E, EM and EF) (p<0.01). Liver total fat (p<0.05) and plasma ALT (P<0.01) were increased by methionine supplementation (EM), implicating fatty liver and liver injury. Liver folate was increased slightly by folate supplementation (EF) (p=0.077). Urinary folate loss was increased 2.3 fold by ethanol consumption (E) and 17.2 fold by folate supplementation (EF), while decreased by methionine supplementation (EM) (p<0.000l). Plasma Hcy was increased 1.9 fold by methionine supplementation (EM) in ethanol-fed mts (p<0.05), which was related with decreased methionine synthase activity (p<0.05). Hepatic SAM/SAH ratio was depressed by methionine supplementation in ethanol-fed mts (EM) (p<0.05). Urinary formininoglutamate (Figlu) excretion after histidine loading was increased by ethanol ingestion and reduced by methionine supplementation (p<0.00l). Based on these data, methionine supplementation appears to accelerate histidine oxidation. In conclusion, dietary supplementation of methionine to ethanol-fed mts exacerbates alcoholic liver injury possibly by complicating sulphur-containing amino acid metabolism, as while it may have beneficial effects on folate and histidine metabolism.

• Biomolecules & Therapeutics
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• 제26권4호
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• pp.399-408
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• 2018

In this study, we examined the molecular and functional characterization of choline uptake in the human esophageal cancer cells. In addition, we examined the influence of various drugs on the transport of [$^3H$]choline, and explored the possible correlation between the inhibition of choline uptake and apoptotic cell death. We found that both choline transporter-like protein 1 (CTL1) and CTL2 mRNAs and proteins were highly expressed in esophageal cancer cell lines (KYSE series). CTL1 and CTL2 were located in the plasma membrane and mitochondria, respectively. Choline uptake was saturable and mediated by a single transport system, which is both $Na^+$-independent and pH-dependent. Choline uptake and cell viability were inhibited by various cationic drugs. Furthermore, a correlation analysis of the potencies of 47 drugs for the inhibition of choline uptake and cell viability showed a strong correlation. Choline uptake inhibitors and choline deficiency each inhibited cell viability and increased caspase-3/7 activity. We conclude that extracellular choline is mainly transported via a CTL1. The functional inhibition of CTL1 by cationic drugs could promote apoptotic cell death. Furthermore, CTL2 may be involved in choline uptake in mitochondria, which is the rate-limiting step in S-adenosylmethionine (SAM) synthesis and DNA methylation. Identification of this CTL1- and CTL2-mediated choline transport system provides a potential new target for esophageal cancer therapy.

• Bulletin of the Korean Chemical Society
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• 제33권1호
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• pp.48-54
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• 2012

The $[Fe_4S_4]^{2+}$ clusters with 2-, 3-, and 4-pyridinemethanethiolate (S2-Pic, S3-Pic, and S4-Pic, respectively) terminal ligands have been synthesized from the ligand substitution reaction of the $(^nBu_4N)_2[Fe_4S_4Cl_4]$ (I) cluster. The new $(^nBu_4N)_2[Fe_4S_4(SR)_4]$ (R = 2-Pic; II, 3-Pic; III, 4-Pic; IV) clusters were characterized by FTIR and UV-Vis spectroscopy. Cluster II was crystallized in the monoclinic space group C2/c with a = 24.530 (5) $\AA$, b = 24.636(4) $\AA$, c = 21.762(4) $\AA$, ${\beta}=103.253(3)^{\circ}$, and Z = 8. The X-ray structure of II showed two unique 2:2 site-differentiated $[Fe_4S_4]^{2+}$ clusters due to the bidentate-mode coordination by 2-pyridinemethanethiolate ligands. Cluster III was crystallized in the same monoclinic space group C2/c with a = 26.0740(18) $\AA$, b = 23.3195(16) $\AA$, c = 22.3720(15) $\AA$, ${\beta}=100.467(2)^{\circ}$, and Z = 8. The 3-pyridinemethanethiolate ligand of III was coordinated to the $[Fe_4S_4]^{2+}$ core as a terminal mode. Cluster IV with 4-pyridinemethanethiolate ligands was found to have a similar structure to the cluster III. Fully reversible $[Fe_4S_4]^{2+}/[Fe_4S_4]^+$ redox waves were observed from all three clusters by cyclic voltammetry measurement. The electrochemical potentials for the $[Fe_4S_4]^{2+}/[Fe_4S_4]^+$ transition decreased in the order of II, III and IV, and the reduction potential changes by the ligands were explained based on the structural differences among the complexes. The complex III was reacted with sulfonium salt of $[PhMeSCH_2-p-C_6H_4CN](BF_4)$ in MeCN to test possible radical-involving reaction as a functional model of the [$Fe_4S_4$]-SAM (S-adenosylmethionine) cofactor. However, the isolated reaction products of 3-pyridinemethanethiolate-p-cyanobenzylsulfide and thioanisole suggested that the reaction followed an ionic mechanism and the products formed from the terminal ligand attack to the sulfonium.

• 대한약리학회지
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• 제32권3호
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• pp.323-334
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• 1996

웅성 모래쥐의 양경동맥을 7분간 폐쇄하여(OBC7) 뇌의 허혈을 유발하여, 해마의 microdialysate내 polyamine 함량과 조직내 polyamine 생합성효소(ornithine decarboxylase: ODC와 S-adenosylmethionine decarboxylase: SAM-DC)의 활성도를 분석하고, 해마의 cresyl violet(CV) 염색과 glial fibrillary acidic protein(GFAP) 면역염색소견들을 관찰하여, 허혈성 해마의 신경손상과 polyamine대사의 연관성을 검토하였다. 1) OBC부하 후, 해마의 dialysate내 polyamine 변동에서, putrescine(PT)은 현저히 증가되었으나, spermidine과 spermine은 다소 감소되는 경향을 보였고, 이에 해마조직내 ODC활성의 현저한 상승과 SAM-DC활성도의 유의한 저학 동반되었다. 2) Difluoromethylornithine(DFMO)는 OBC에 의한 PT증가와 ODC활성도 상승을 유의하게 억제하였으나, methylglyoxal bis(guanylhydrazone)(MGBG)는 각각 다소 억제하는 경향을 보였다. 3) OBC부하 7일후에 관찰한 조직소견에서, 해마의 CA1 부위의 유의한 신경손상이 유도되었으나 CA3와 dentate gyrus 부위에는 미약한 손상만 보였으며, GFAP 양성반응도 CA1 부위에서만 유의한 증가를 보였다. 이같은 소견들은 DFMO에 의하여 크게 영향을 받지 않았으나 MGBG에 의하여 유의하게 억제되었다. 이상의 성적들은 해마 polyamine의 과도한 허혈성 증가가 허혈성 신경손상에 관여할 수 있으나, 한편으로 polyamine대사의 과도한 억제도 허혈성 신경손상을 악화시킬 수 있으며, 허혈성뇌손상에 대한 MGBG의 보호작용은 polyamine 대사보다는 다른 작용에 매개되는 것으로 사료된다. 가장 많았고 습진과 소양증이 각각 19.5%였다. 30대 환자군에서는 습진이 30.3%였고 소양증이 19.1%였다. 40대에서는 소양증이 28.8%였고 지루성 피부염이 16.0%였다. 50대에서는 습진이 26.7%였고 60세 이상의 그룹에서는 습진이 50.0%, 바이러스 질환과 소양증이 각각 15.0%였다. 이들 결과는 통계적으로 유의한 차이를 보였다($X^2\;=\;74.995^{**}$, p = .001, df : 40). 결론 : 상대적으로 50세 이상 연령충의 대상 환자 수가 많지 않은 원인은 여리 가지 분석이 가능할 것으로 생각된다. 연령에 따라 피부질환이 다른 양상을 보임을 고려할 때 노인층의 신체적, 경제적, 사회적 특성을 이해하고 이에 따른 효과적이고 사회-경제학적 부담을 최소할 수 있는 한방 치료 영역의 확대 및 개발이 필요할 것으로 사려된다.주식 및 후식의 선호도에 남녀 간에 차이를 보이고 있으므로 단체급식을 위한 식단 작성 시 이러한 차이를 고려할 필요가 있으며 대학생 및 성인을 대상으로 한 영양 교육을 실시할 때 이러한 차이를 고려한 신생활 교정프로그램을 작성할 필요가 있다고 사료된다.$은 대조군에 비해 발아에 의하여 세포독성 효과를 증가되었지만, MCF-7와 Caco-2에 대한 항암효과는 없음을 알 수 있었다.것으로 사료된다.높게 인식할수록 재방문의도 및 추천의도가 커지는 것을 알 수 있었다. 대학교 급식소 운영주체에 대한 소비자 인지도 조사결과 향후 대학교 급식소를 운영하는 위탁급식 전문업체의 경우 그들의 브랜드를 알리기 위한 홍보전략이 절실히 필요함을 알 수 있었으며, 최근고객감소로 인하여 다양한 급식운영 마케팅전략을 수립하고 있는 단체급식 운영자들은 재방문 및 추천의도의 선행요건이 급식서비스 품질에 대한 소비자의 인식임을 명심하여 가장 기본이 되는 급식서비스 품질수준을 향상시키기 위하여 노력하여야 할 것이다.