[논문]Cloning, Expression, and Characterization of a Thermostable GH51 <tex>${\\alpha}-\\small{L}$</tex>-Arabinofuranosidase from Paenibacillus sp. DG-22

Lee, Sun Hwa; Lee, Yong-Eok

doi:10.4014/jmb.1308.08078

[국내논문] Cloning, Expression, and Characterization of a Thermostable GH51 ${\\alpha}-\\small{L}$-Arabinofuranosidase from Paenibacillus sp. DG-22 원문보기

Journal of microbiology and biotechnology, v.24 no.2, 2014년, pp.236 - 244

Lee, Sun Hwa (Department of Biotechnology, Dongguk University) , Lee, Yong-Eok (Department of Biotechnology, Dongguk University)

Abstract ▼ AI-Helper

The gene encoding ${\alpha}-\small{L}$-arabinofuranosidase (AFase) from Paenibacillus sp. DG-22 was cloned, sequenced, and expressed in Escherichia coli. The AFase gene (abfA) comprises a 1,509 bp open reading frame encoding 502 amino acids with a molecular mass of 56,520 daltons. The deduced amino acid sequence of the gene shows that AbfA is an enzyme consisting of only a catalytic domain, and that the enzyme has significant similarity to AFases classified into the family 51 of the glycosyl hydrolases. abfA was subcloned into the pQE60 expression vector to fuse it with a six-histidine tag and the recombinant AFase (rAbfA) was purified to homogeneity. The specific activity of the recombinant enzyme was 96.7 U/mg protein. Determination of the apparent molecular mass by gel-filtration chromatography indicated that AbfA has a tetrameric structure. The optimal pH and temperature of the enzyme were 6.0 and $60^{\circ}C$, respectively. The enzyme activity was completely inhibited by 1 mM $HgCl_2$. rAbfA was active only towards p-nitrophephenyl ${\alpha}-\small{L}$-arabinofuranoside and exhibited $K_m$ and $V_{max}$ values of 3.5 mM and 306.1 U/mg, respectively. rAbfA showed a synergistic effect in combination with endoxylanase on the degradation of oat spelt xylan and wheat arabinoxylan.

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이론/모형

DNA sequencing was conducted at the Genotech DNA sequencing facility (Daejeon, Korea) by automated sequencing using the dideoxynucleotide chain termination method. The nucleotide sequence was analyzed using the National Center for Biotechnology Information (NCBI) Open Reading Frame (ORF) Finder tool.
DNA sequencing was conducted at the Genotech DNA sequencing facility (Daejeon, Korea) by automated sequencing using the dideoxynucleotide chain termination method. The nucleotide sequence was analyzed using the National Center for Biotechnology Information (NCBI) Open Reading Frame (ORF) Finder tool. The signal peptide in the deduced amino acid sequence was predicted by the SignalP 4.
The protein content was determined by the Bradford method [6] with Protein Assay reagent (Bio-Rad) using bovine serum albumin as the standard. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed on a 15% running gel [17] and resolved proteins were visualized by staining with Coomassie Brilliant Blue R-250 (Sigma).
aReducing sugar was measured using the DNS method [23].

참고문헌 (37)

Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25: 3389-3402.

상세보기
Alvira P, Negro MJ, Ballesteros M. 2011. Effect of endoxylanase and $\alpha$ -L-arabinofuranosidase supplementation on the enzymatic hydrolysis of steam exploded wheat straw. Bioresour. Technol. 102: 4552-4558.

상세보기
Arti D, Park JM, Jung TY, Song HN, Jang MU, Han NS, et al. 2012. Structural analysis of $\alpha$ -L-arabinofuranosidase from Thermotoga maritima reveals characteristics for thermostability and substrate specificity. J. Microbiol. Biotechnol. 22: 1724- 1730.

원문보기 상세보기
Bastawde KB. 1992. Xylan structure, microbial xylanases, and their mode of action. World J. Microbiol. Biotechnol. 8: 353-368.

상세보기
Biely P. 1985. Microbial xylanolytic systems. Trends Biotechnol. 3: 286-290.

상세보기
Bradford M M. 1976. Arapid and s ensitive m ethod for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-256.

상세보기
Canakei S, Belduz AO, Saha BC, Yasar A, Ayaz FA, Yayli N. 2007. Purification and characterization of a highly thermostable $\alpha$ -L-arabinofuranosidase from Geobacillus caldoxylolyticus TK4. Appl. Microbiol. Biotechnol. 75: 813-820.

상세보기
Debeche T, Cummings N, Connerton I, Debeire P, O'Donohue MJ. 2000. Genetic and biochemical characterization of a highly thermostable $\alpha$ -L-arabinofuranosidase from Thermobacillus xylanilyticus. Appl. Environ. Microbiol. 66: 1734-1736.

상세보기
Degrassi G, Vindigni A, Venturi V. 2003. Thermostable $\alpha$ -arabinofuranosidase from xylanolytic Bacillus pumilus: purification and characterization. J. Biotechnol. 101: 69-79.

상세보기
Gilead S, Shoham Y. 1995. Purification and characterization of $\alpha$ -L-arabinofuranosidase from Bacillus stearothermophilus T-6. Appl. Environ. Microbiol. 61: 170-174.

상세보기
Gouet P, Robert X, Courcelle E. 2003. ESPript/ENDscript: extracting and rendering sequence and 3D information from atomic structures of proteins. Nucleic Acids Res. 31: 3320- 3323.

상세보기
Henrissat B, Bairoch A. 1996. Updating the sequence-based classification of glycosyl hydrolases. Biochem. J. 316: 695-696.

상세보기
Hovel K, Shallom D, Niefind K, Belakhov V, Shoham G, Baasov T, et al. 2003. Crystal structure and snapshots along the reaction pathway of a family 51 $\alpha$ -L-arabinofuranosidase. EMBO J. 22: 4922-4932.

상세보기
Kaji A. 1984. L-Arabinosidases. Adv. Carbohydr. Chem. Biochem. 42: 383-394.
Knob A, Carmona EC. 2010. Purification and characterization of two extracellular xylanases from Penicillium sclerotiorum: a novel acidophilic xylanase. Appl. Biochem. Biotechnol. 162: 429-443.

상세보기
Kulkarni N, Shendye A, Rao M. 1999. Molecular and biotechnological aspects of xylanases. FEMS Microbiol. Rev. 23: 411-456.

상세보기
Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.

상세보기
Lee TH, Lee YE. 2007. Cloning, sequencing and expression of the gene encoding a thermostable $\beta$ -xylosidase from Paenibacillus sp. DG-22. J. Life Sci. 17: 1197-1203.

원문보기 상세보기
Lee TH, L im P O, L ee YE. 2007 . Cloning, characterization, and expression of xylanase A gene from Paenibacillus sp. DG-22 in Escherichia coli. J. Microbiol. Biotechnol. 17: 29-36.

원문보기 상세보기
Lee YE. 2004. Isolation and characterization of thermostable xylanase producing Paenibacillus sp. DG-22. Kor. J. Microbiol. Biotechnol. 32: 22-28.

상세보기
Lee YE, Lim PO. 2004. Purification and characterization of two thermostable xylanases from Paenibacillus sp. DG-22. J. Microbiol. Biotechnol. 14: 1014-1021.

원문보기 상세보기
Marmur J. 1961. A procedure for the isolation of deoxyribonucleic acid from microorganisms. J. Mol. Biol. 3: 208-218.

상세보기
Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugars. Anal. Chem. 31: 426-428.

상세보기
Morana A, Paris O, Maurelli L, Rossi M, Cannio R. 2007. Gene cloning and expression in Escherichia coli of a bifunctional $\beta$ -D-xylosidase/ $\alpha$ -L-arabinosidase from Sulfolobus solfataricus involved in xylan degradation. Extremophiles 11: 123-132.

상세보기
Numan MT, Bhosle NB. 2006. $\alpha$ -L-Arabinofuranosidase: the potential applications in biotechnology. J. Ind. Microbiol. Biotechnol. 33: 247-260.

상세보기
Paes G, Skov LK, O'Donohue MJ, Remond C, Kastrup JS, Gajhede M, Mirza O. 2008. The structure of the complex between a branched pentasaccharide and Thermobacillus xylanilyticus GH-51 arabinofuranosidase reveals xylan-binding determinants and induced fit. Biochemistry 47: 7441-7451.

상세보기
Pei J, Shao W. 2008. Purification and characterization of an extracellular $\alpha$ -L-arabinosidase from a novel isolate Bacillus pumilus ARA and its over-expression in Escherichia coli. Appl. Microbiol. Biotechnol. 78: 115-121.

상세보기
Petersen TN, Brunak S, Heijne G, Nielsen H. 2011. SignalP 4,0: discriminating signal peptides from transmembrane regions. Nat. Methods 8: 785-786.

상세보기
Raweesri P, Riangrungrojana P, Pinphanichakarn P. 2008. $\alpha$ - L-Arabinofuranosidase from Streptomyces sp. PC22: purification, characterization and its synergistic action with xylanolytic enzymes in the degradation of xylan and agricultural residues. Bioresour. Technol. 99: 8981-8986.

상세보기
Saha BC. 2000. $\alpha$ -L-Arabinofuranosidase: biochemistry, molecular biology and application in biotechnology. Biotechnol. Adv. 18: 403-423.

상세보기
Shi P, Chen X, Meng K, Huang H, Bai Y, Luo H, et al. 2013. Distinct actions by Paenibacillus sp. strain E18 $\alpha$ -Larabinofuranosidases and xylanase in xylan degradation. Appl. Environ. Microbiol. 79: 1990-1995.

상세보기
Sievers F, Wilm A, Dineen DG, Gibson TJ, Karplus K, Li W, et al. 2011. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol. Syst. Biol. 7: 539
Souza TACB, Santos CR, Souza AR, Oldiges DP, Ruller R, Prade RA, et al. 2011. Structure of a novel thermostable GH51 $\alpha$ -L-arabinofuranosidase from Thermotoga petrophila RKU-1. Protein Sci. 20: 1632-1637.

상세보기
Sunna A, Antranikian G. 1997. Xylanolytic enzymes from fungi and bacteria. Crit. Rev. Biotechnol. 17: 39-67.

상세보기
Taylor EJ, Smith NL, Turkenburg JP, D'Souza S, Gilbert HJ, Davies GJ. 2006. Structural insight into the ligand specificity of a thermostable family 51 arabinofuranosidase, Araf51, from Clostridium thermocellum. Biochem. J. 395: 31-37.

상세보기
Ward OP, Moo-Young M. 1989. Enzymatic degradation of cell wall and related plant polysaccharides. Crit. Rev. Biotechnol. 8: 237-274.

상세보기
Wiegel VM, Lorenz WW. 2000. Cloning, sequencing, and characterization of the bifunctional xylosidase-arabinosidase from the anaerobic thermophile Thermoanaerobacter ethanolicus. Gene 247: 137-143.

상세보기

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