메뉴 건너뛰기




Volumn 78, Issue 17, 2012, Pages 6113-6120

MhbT Is a specific transporter for 3-hydroxybenzoate uptake by gram-negative bacteria

Author keywords

[No Author keywords available]

Indexed keywords

AMINO ACID RESIDUES; CYTOPLASMIC LOOPS; CYTOPLASMIC MEMBRANE; FLUORESCENT PROTEIN; FUMARATES; FUSION PROTEINS; GENE CLUSTERS; GENTISATE; GRAM-NEGATIVE BACTERIA; HYPOTHETICAL PROTEIN; KLEBSIELLA PNEUMONIAE; METABOLIC INTERMEDIATES; MUTANT PROTEINS; PSEUDOMONAS PUTIDA; PYRUVATES; SITE DIRECTED MUTAGENESIS; SIX GENES;

EID: 84866157964     PISSN: 00992240     EISSN: 10985336     Source Type: Journal    
DOI: 10.1128/AEM.01511-12     Document Type: Article
Times cited : (24)

References (50)
  • 2
    • 0017184389 scopus 로고
    • A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
    • Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254.
    • (1976) Anal. Biochem. , vol.72 , pp. 248-254
    • Bradford, M.M.1
  • 3
    • 33947372628 scopus 로고    scopus 로고
    • Genome-wide investigation of aromatic acid transporters in Corynebacterium glutamicum
    • Chaudhry MT, et al. 2007. Genome-wide investigation of aromatic acid transporters in Corynebacterium glutamicum. Microbiology 153:857-865.
    • (2007) Microbiology , vol.153 , pp. 857-865
    • Chaudhry, M.T.1
  • 5
    • 0030983977 scopus 로고    scopus 로고
    • benK encodes a hydrophobic permease-like protein involved in benzoate degradation by Acinetobacter sp. strain ADP1
    • Collier LS, Nichols NN, Neidle EL. 1997. benK encodes a hydrophobic permease-like protein involved in benzoate degradation by Acinetobacter sp. strain ADP1. J. Bacteriol. 179:5943-5946.
    • (1997) J. Bacteriol. , vol.179 , pp. 5943-5946
    • Collier, L.S.1    Nichols, N.N.2    Neidle, E.L.3
  • 6
    • 0034612342 scopus 로고    scopus 로고
    • One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products
    • Datsenko KA, Wanner BL. 2000. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. U. S. A. 97:6640-6645.
    • (2000) Proc. Natl. Acad. Sci. U. S. A. , vol.97 , pp. 6640-6645
    • Datsenko, K.A.1    Wanner, B.L.2
  • 7
    • 0027390501 scopus 로고
    • Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes
    • de Lorenzo V, Eltis L, Kessler B, Timmis KN. 1993. Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes. Gene 123:17-24.
    • (1993) Gene , vol.123 , pp. 17-24
    • de Lorenzo, V.1    Eltis, L.2    Kessler, B.3    Timmis, K.N.4
  • 8
    • 0036175122 scopus 로고    scopus 로고
    • + symporter family of permeases are required for 4-hydroxybenzoate transport by PcaK from Pseudomonas putida
    • + symporter family of permeases are required for 4-hydroxybenzoate transport by PcaK from Pseudomonas putida. J. Bacteriol. 184:1444-1448.
    • (2002) J. Bacteriol. , vol.184 , pp. 1444-1448
    • Ditty, J.L.1    Harwood, C.S.2
  • 9
    • 0032864202 scopus 로고    scopus 로고
    • Conserved cytoplasmic loops are important for both the transport and chemotaxis functions of PcaK, a protein from Pseudomonas putida with 12 membrane-spanning regions
    • Ditty JL, Harwood CS. 1999. Conserved cytoplasmic loops are important for both the transport and chemotaxis functions of PcaK, a protein from Pseudomonas putida with 12 membrane-spanning regions. J. Bacteriol. 181:5068-5074.
    • (1999) J. Bacteriol. , vol.181 , pp. 5068-5074
    • Ditty, J.L.1    Harwood, C.S.2
  • 10
    • 0021531509 scopus 로고
    • The genetic complexity of nitrogen fixation. The ninth Fleming lecture
    • Dixon RA. 1984. The genetic complexity of nitrogen fixation. The ninth Fleming lecture. J. Gen. Microbiol. 130:2745-2755.
    • (1984) J. Gen. Microbiol. , vol.130 , pp. 2745-2755
    • Dixon, R.A.1
  • 11
    • 0015212112 scopus 로고
    • Transfer of nitrogen-fixation genes by conjugation in Klebsiella pneumoniae
    • Dixon RA, Postgate JR. 1971. Transfer of nitrogen-fixation genes by conjugation in Klebsiella pneumoniae. Nature 234:47-48.
    • (1971) Nature , vol.234 , pp. 47-48
    • Dixon, R.A.1    Postgate, J.R.2
  • 12
    • 27744568351 scopus 로고    scopus 로고
    • Molecular and biochemical characterization of the xlnD-encoded 3-hydroxybenzoate 6-hydroxylase involved in the degradation of 2,5-xylenol via the gentisate pathway in Pseudomonas alcaligenes NCIMB 9867
    • Gao X, Tan CL, Yeo CC, Poh CL. 2005. Molecular and biochemical characterization of the xlnD-encoded 3-hydroxybenzoate 6-hydroxylase involved in the degradation of 2,5-xylenol via the gentisate pathway in Pseudomonas alcaligenes NCIMB 9867. J. Bacteriol. 187:7696-7702.
    • (2005) J. Bacteriol. , vol.187 , pp. 7696-7702
    • Gao, X.1    Tan, C.L.2    Yeo, C.C.3    Poh, C.L.4
  • 14
    • 0029083173 scopus 로고
    • + antiport catalyzed by the Bacillus subtilis TetA(L) transporter expressed in Escherichia coli
    • + antiport catalyzed by the Bacillus subtilis TetA(L) transporter expressed in Escherichia coli. J. Bacteriol. 177:4557-4561.
    • (1995) J. Bacteriol. , vol.177 , pp. 4557-4561
    • Guffanti, A.A.1    Krulwich, T.A.2
  • 15
    • 0028172818 scopus 로고
    • Identification of the pcaRKF gene cluster from Pseudomonas putida: involvement in chemotaxis, biodegradation, and transport of 4-hydroxybenzoate
    • Harwood CS, Nichols NN, Kim MK, Ditty JL, Parales RE. 1994. Identification of the pcaRKF gene cluster from Pseudomonas putida: involvement in chemotaxis, biodegradation, and transport of 4-hydroxybenzoate.J. Bacteriol. 176:6479-6488.
    • (1994) J. Bacteriol. , vol.176 , pp. 6479-6488
    • Harwood, C.S.1    Nichols, N.N.2    Kim, M.K.3    Ditty, J.L.4    Parales, R.E.5
  • 16
    • 0037373068 scopus 로고    scopus 로고
    • Structural model for 12-helix transporters belonging to the major facilitator superfamily
    • Hirai T, Heymann JA, Maloney PC, Subramaniam S. 2003. Structural model for 12-helix transporters belonging to the major facilitator superfamily. J. Bacteriol. 185:1712-1718.
    • (2003) J. Bacteriol. , vol.185 , pp. 1712-1718
    • Hirai, T.1    Heymann, J.A.2    Maloney, P.C.3    Subramaniam, S.4
  • 17
    • 1642338888 scopus 로고    scopus 로고
    • Novel pathway of salicylate degradation by Streptomyces sp. strain WA46
    • Ishiyama D, Vujaklija D, Davies J. 2004. Novel pathway of salicylate degradation by Streptomyces sp. strain WA46. Appl. Environ. Microbiol. 70:1297-1306.
    • (2004) Appl. Environ. Microbiol. , vol.70 , pp. 1297-1306
    • Ishiyama, D.1    Vujaklija, D.2    Davies, J.3
  • 18
    • 0020046839 scopus 로고
    • Excision and integration of degradative pathway genes from TOL plasmid pWW0
    • Jeenes DJ, Williams PA. 1982. Excision and integration of degradative pathway genes from TOL plasmid pWW0. J. Bacteriol. 150:188-194.
    • (1982) J. Bacteriol. , vol.150 , pp. 188-194
    • Jeenes, D.J.1    Williams, P.A.2
  • 19
    • 0030999256 scopus 로고    scopus 로고
    • Suppressor analysis of mutations in the loop 2-3 motif of lactose permease: evidence that glycine-64 is an important residue for conformational changes
    • Jessen-Marshall AE, Parker NJ, Brooker RJ. 1997. Suppressor analysis of mutations in the loop 2-3 motif of lactose permease: evidence that glycine-64 is an important residue for conformational changes. J. Bacteriol. 179:2616-2622.
    • (1997) J. Bacteriol. , vol.179 , pp. 2616-2622
    • Jessen-Marshall, A.E.1    Parker, N.J.2    Brooker, R.J.3
  • 20
    • 0025130003 scopus 로고
    • Catabolism of 3-hydroxybenzoate by the gentisate pathway in Klebsiella pneumoniae M5a1
    • Jones DC, Cooper RA. 1990. Catabolism of 3-hydroxybenzoate by the gentisate pathway in Klebsiella pneumoniae M5a1. Arch. Microbiol. 154:489-495.
    • (1990) Arch. Microbiol. , vol.154 , pp. 489-495
    • Jones, D.C.1    Cooper, R.A.2
  • 21
    • 0021783513 scopus 로고
    • The proton motive force in bacteria: a critical assessment of methods
    • Kashket ER. 1985. The proton motive force in bacteria: a critical assessment of methods. Annu. Rev. Microbiol. 39:219-242.
    • (1985) Annu. Rev. Microbiol. , vol.39 , pp. 219-242
    • Kashket, E.R.1
  • 22
    • 0023715368 scopus 로고
    • Improved broadhost-range plasmids for DNA cloning in gram-negative bacteria
    • Keen NT, Tamaki S, Kobayashi D, Trollinger D. 1988. Improved broadhost-range plasmids for DNA cloning in gram-negative bacteria. Gene 70:191-197.
    • (1988) Gene , vol.70 , pp. 191-197
    • Keen, N.T.1    Tamaki, S.2    Kobayashi, D.3    Trollinger, D.4
  • 23
    • 53849119555 scopus 로고    scopus 로고
    • Ins and outs of major facilitator superfamily antiporters
    • Law CJ, Maloney PC, Wang DN. 2008. Ins and outs of major facilitator superfamily antiporters. Annu. Rev. Microbiol. 62:289-305.
    • (2008) Annu. Rev. Microbiol. , vol.62 , pp. 289-305
    • Law, C.J.1    Maloney, P.C.2    Wang, D.N.3
  • 24
    • 0031948730 scopus 로고    scopus 로고
    • The tfdK gene product facilitates uptake of 2,4-dichlorophenoxyacetate by Ralstonia eutropha JMP134(pJP4)
    • Leveau JH, Zehnder AJ, van der Meer JR. 1998. The tfdK gene product facilitates uptake of 2,4-dichlorophenoxyacetate by Ralstonia eutropha JMP134(pJP4). J. Bacteriol. 180:2237-2243.
    • (1998) J. Bacteriol. , vol.180 , pp. 2237-2243
    • Leveau, J.H.1    Zehnder, A.J.2    van der Meer, J.R.3
  • 25
    • 70450224489 scopus 로고    scopus 로고
    • MhbR, a LysR-type regulator involved in 3-hydroxybenzoate catabolism via gentisate in Klebsiella pneumoniae M5a1
    • Lin LX, Liu H, Zhou NY. 2010. MhbR, a LysR-type regulator involved in 3-hydroxybenzoate catabolism via gentisate in Klebsiella pneumoniae M5a1. Microbiol. Res. 165:66-74.
    • (2010) Microbiol. Res. , vol.165 , pp. 66-74
    • Lin, L.X.1    Liu, H.2    Zhou, N.Y.3
  • 26
    • 13444282320 scopus 로고    scopus 로고
    • Arg169 is essential for catalytic activity of 3-hydroxybenzoate 6-hydroxylase from Klebsiella pneumoniae M5a1
    • Liu DQ, Liu H, Gao XL, Leak DJ, Zhou NY. 2005. Arg169 is essential for catalytic activity of 3-hydroxybenzoate 6-hydroxylase from Klebsiella pneumoniae M5a1. Microbiol. Res. 160:53-59.
    • (2005) Microbiol. Res. , vol.160 , pp. 53-59
    • Liu, D.Q.1    Liu, H.2    Gao, X.L.3    Leak, D.J.4    Zhou, N.Y.5
  • 27
    • 15044356850 scopus 로고    scopus 로고
    • A new isolate of Pseudomonas stutzerithat degrades 2-chloronitrobenzene
    • Liu H, Wang SJ, Zhou NY. 2005. A new isolate of Pseudomonas stutzerithat degrades 2-chloronitrobenzene. Biotechnol. Lett. 27:275-278.
    • (2005) Biotechnol. Lett. , vol.27 , pp. 275-278
    • Liu, H.1    Wang, S.J.2    Zhou, N.Y.3
  • 28
    • 79954659728 scopus 로고    scopus 로고
    • Functional characterization of a gene cluster involved in gentisate catabolism in Rhodococcus s. strain NCIMB 12038
    • Liu TT, et al. 2011. Functional characterization of a gene cluster involved in gentisate catabolism in Rhodococcus s. strain NCIMB 12038. Appl. Microbiol. Biotechnol. 90:671-678.
    • (2011) Appl. Microbiol. Biotechnol. , vol.90 , pp. 671-678
    • Liu, T.T.1
  • 30
    • 30944465384 scopus 로고    scopus 로고
    • Site-directed mutagenesis of gentisate 1,2-dioxygenases from Klebsiella pneumoniae M5a1 and Ralstonia sp. strain U2
    • Luo S, Liu DQ, Liu H, Zhou NY. 2006. Site-directed mutagenesis of gentisate 1,2-dioxygenases from Klebsiella pneumoniae M5a1 and Ralstonia sp. strain U2. Microbiol. Res. 161:138-144.
    • (2006) Microbiol. Res. , vol.161 , pp. 138-144
    • Luo, S.1    Liu, D.Q.2    Liu, H.3    Zhou, N.Y.4
  • 31
    • 0028790371 scopus 로고
    • Uptake of benzoic acid and chloro-substituted benzoic acids by Alcaligenes denitrificans BRI 3010 and BRI 6011
    • Miguez CB, Greer CW, Ingram JM, Macleod RA. 1995. Uptake of benzoic acid and chloro-substituted benzoic acids by Alcaligenes denitrificans BRI 3010 and BRI 6011. Appl. Environ. Microbiol. 61:4152-4159.
    • (1995) Appl. Environ. Microbiol. , vol.61 , pp. 4152-4159
    • Miguez, C.B.1    Greer, C.W.2    Ingram, J.M.3    Macleod, R.A.4
  • 32
    • 33144463952 scopus 로고    scopus 로고
    • Cloning and functional characterization of the styE gene, involved in styrene transport in Pseudomonas putida CA-3
    • Mooney A, O'Leary ND, Dobson AD. 2006. Cloning and functional characterization of the styE gene, involved in styrene transport in Pseudomonas putida CA-3. Appl. Environ. Microbiol. 72:1302-1309.
    • (2006) Appl. Environ. Microbiol. , vol.72 , pp. 1302-1309
    • Mooney, A.1    O'Leary, N.D.2    Dobson, A.D.3
  • 33
    • 0030818839 scopus 로고    scopus 로고
    • PcaK, a high-affinity permease for the aromatic compounds 4-hydroxybenzoate and protocatechuate from Pseudomonas putida
    • Nichols NN, Harwood CS. 1997. PcaK, a high-affinity permease for the aromatic compounds 4-hydroxybenzoate and protocatechuate from Pseudomonas putida. J. Bacteriol. 179:5056-5061.
    • (1997) J. Bacteriol. , vol.179 , pp. 5056-5061
    • Nichols, N.N.1    Harwood, C.S.2
  • 35
    • 34548233702 scopus 로고    scopus 로고
    • Molecular and biochemical characterization of 3-hydroxybenzoate 6-hydroxylase from Polaromonas naphthalenivorans CJ2
    • Park M, et al. 2007. Molecular and biochemical characterization of 3-hydroxybenzoate 6-hydroxylase from Polaromonas naphthalenivorans CJ2. Appl. Environ. Microbiol. 73:5146-5152.
    • (2007) Appl. Environ. Microbiol. , vol.73 , pp. 5146-5152
    • Park, M.1
  • 36
    • 0029682161 scopus 로고    scopus 로고
    • In vitro recombination and mutagenesis by overlap extension PCR
    • Pogulis RJ, Vallejo AN, Pease LR. 1996. In vitro recombination and mutagenesis by overlap extension PCR. Methods Mol. Biol. 57:167-176.
    • (1996) Methods Mol. Biol. , vol.57 , pp. 167-176
    • Pogulis, R.J.1    Vallejo, A.N.2    Pease, L.R.3
  • 37
    • 1842611517 scopus 로고    scopus 로고
    • Experimentally based topology models for E. coli inner membrane proteins
    • Rapp M, et al. 2004. Experimentally based topology models for E. coli inner membrane proteins. Protein Sci. 13:937-945.
    • (2004) Protein Sci. , vol.13 , pp. 937-945
    • Rapp, M.1
  • 38
    • 0029801452 scopus 로고    scopus 로고
    • In vitro formation of a catabolic plasmid carrying Klebsiella pneumoniae DNA that allows growth of Escherichia coli K-12 on 3-hydroxybenzoate
    • Robson ND, Parrott S, Cooper RA. 1996. In vitro formation of a catabolic plasmid carrying Klebsiella pneumoniae DNA that allows growth of Escherichia coli K-12 on 3-hydroxybenzoate. Microbiology 142:2115-2120.
    • (1996) Microbiology , vol.142 , pp. 2115-2120
    • Robson, N.D.1    Parrott, S.2    Cooper, R.A.3
  • 40
    • 22144441744 scopus 로고    scopus 로고
    • Functional identification of novel genes involved in the glutathione-independent gentisate pathway in Corynebacterium glutamicum
    • Shen XH, Jiang CY, Huang Y, Liu ZP, Liu SJ. 2005. Functional identification of novel genes involved in the glutathione-independent gentisate pathway in Corynebacterium glutamicum. Appl. Environ. Microbiol. 71:3442-3452.
    • (2005) Appl. Environ. Microbiol. , vol.71 , pp. 3442-3452
    • Shen, X.H.1    Jiang, C.Y.2    Huang, Y.3    Liu, Z.P.4    Liu, S.J.5
  • 41
    • 0037199152 scopus 로고    scopus 로고
    • Metabolic engineering of Pseudomonas putida for the utilization of parathion as a carbon and energy source
    • Walker AW, Keasling JD. 2002. Metabolic engineering of Pseudomonas putida for the utilization of parathion as a carbon and energy source. Biotechnol. Bioeng. 78:715-721.
    • (2002) Biotechnol. Bioeng. , vol.78 , pp. 715-721
    • Walker, A.W.1    Keasling, J.D.2
  • 42
    • 0023663328 scopus 로고
    • Pseudomonas cepacia 3-hydroxybenzoate 6-hydroxylase: induction, purification, and characterization
    • Wang LH, Hamzah RY, Yu YM, Tu SC. 1987. Pseudomonas cepacia 3-hydroxybenzoate 6-hydroxylase: induction, purification, and characterization. Biochemistry 26:1099-1104.
    • (1987) Biochemistry , vol.26 , pp. 1099-1104
    • Wang, L.H.1    Hamzah, R.Y.2    Yu, Y.M.3    Tu, S.C.4
  • 43
    • 0016197816 scopus 로고
    • Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid
    • Williams PA, Murray K. 1974. Metabolism of benzoate and the methylbenzoates by Pseudomonas putida (arvilla) mt-2: evidence for the existence of a TOL plasmid. J. Bacteriol. 120:416-423.
    • (1974) J. Bacteriol. , vol.120 , pp. 416-423
    • Williams, P.A.1    Murray, K.2
  • 44
    • 34548552746 scopus 로고    scopus 로고
    • Molecular characterization of a novel ortho-nitrophenol catabolic gene cluster in Alcaligenes sp. strain NyZ215
    • Xiao Y, Zhang JJ, Liu H, Zhou NY. 2007. Molecular characterization of a novel ortho-nitrophenol catabolic gene cluster in Alcaligenes sp. strain NyZ215. J. Bacteriol. 189:6587-6593.
    • (2007) J. Bacteriol. , vol.189 , pp. 6587-6593
    • Xiao, Y.1    Zhang, J.J.2    Liu, H.3    Zhou, N.Y.4
  • 45
    • 33745135388 scopus 로고    scopus 로고
    • Heterologous expression and localization of gentisate transporter Ncg12922 from Corynebacterium glutamicum ATCC 13032
    • Xu Y, Yan DZ, Zhou NY. 2006. Heterologous expression and localization of gentisate transporter Ncg12922 from Corynebacterium glutamicum ATCC 13032. Biochem. Biophys. Res. Commun. 346:555-561.
    • (2006) Biochem. Biophys. Res. Commun. , vol.346 , pp. 555-561
    • Xu, Y.1    Yan, D.Z.2    Zhou, N.Y.3
  • 46
    • 78650387382 scopus 로고    scopus 로고
    • Purification and characterization of the ncgl2923-encoded 3-hydroxybenzoate 6-hydroxylase from Corynebacterium glutamicum
    • Yang YF, Zhang JJ, Wang SH, Zhou NY. 2010. Purification and characterization of the ncgl2923-encoded 3-hydroxybenzoate 6-hydroxylase from Corynebacterium glutamicum. J. Basic Microbiol. 50:599-604.
    • (2010) J. Basic Microbiol. , vol.50 , pp. 599-604
    • Yang, Y.F.1    Zhang, J.J.2    Wang, S.H.3    Zhou, N.Y.4
  • 47
    • 77955552309 scopus 로고    scopus 로고
    • Characterization of catabolic meta-nitrophenol nitroreductase from Cupriavidus necator JMP134
    • Yin Y, et al. 2010. Characterization of catabolic meta-nitrophenol nitroreductase from Cupriavidus necator JMP134. Appl. Microbiol. Biotechnol. 87:2077-2085.
    • (2010) Appl. Microbiol. Biotechnol. , vol.87 , pp. 2077-2085
    • Yin, Y.1
  • 48
    • 52649129267 scopus 로고    scopus 로고
    • Identification and characterization of the dicarboxylate uptake system DccT in Corynebacterium glutamicum
    • Youn JW, Jolkver E, Kramer R, Marin K, Wendisch VF. 2008. Identification and characterization of the dicarboxylate uptake system DccT in Corynebacterium glutamicum. J. Bacteriol. 190:6458-6466.
    • (2008) J. Bacteriol. , vol.190 , pp. 6458-6466
    • Youn, J.W.1    Jolkver, E.2    Kramer, R.3    Marin, K.4    Wendisch, V.F.5
  • 49
    • 33748299619 scopus 로고    scopus 로고
    • Discovery and investigation of a new, second triose phosphate isomerase in Klebsiella pneumoniae
    • Zheng P, Sun J, van den Heuvel J, Zeng AP. 2006. Discovery and investigation of a new, second triose phosphate isomerase in Klebsiella pneumoniae. J. Biotechnol. 125:462-473.
    • (2006) J. Biotechnol. , vol.125 , pp. 462-473
    • Zheng, P.1    Sun, J.2    van den Heuvel, J.3    Zeng, A.P.4
  • 50
    • 0035167509 scopus 로고    scopus 로고
    • nag genes of Ralstonia (formerly Pseudomonas) sp. strain U2 encoding enzymes for gentisate catabolism
    • Zhou NY, Fuenmayor SL, Williams PA. 2001. nag genes of Ralstonia (formerly Pseudomonas) sp. strain U2 encoding enzymes for gentisate catabolism. J. Bacteriol. 183:700-708.
    • (2001) J. Bacteriol. , vol.183 , pp. 700-708
    • Zhou, N.Y.1    Fuenmayor, S.L.2    Williams, P.A.3


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.