용인 함박산 토양에서 분리한 Paenibacillus sp. HX-1의 동정과 endo-β-1,4-xylanase 생산 증가를 위한 배지최적화
Enhanced Production of Endo-β-1,4-xylanase from Paenibacillus sp. HX-1 Newly Isolated from Soil Samples at Hambak Mountain in Yongin city, Korea.
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균주 HX-1은 토양샘플로부터 분리된 자일라네이즈 생산 미생물로서 16S rRNA 유전자 염기서열 분석과 이를 이용한 phylogenetic tree 제작을 통하여 Paenibacillus 속의 한종으로 동정되었다. 그러나 HX-1 균주가 계통발생적 연관관계가 높은 기존에 알려진 표준군주들과는 상당히 다른 생리적-생화학적 특성을 나타내는 사실로부터 HX-1이 신아종일 것으로 판단하고, Paenibacillus sp. HX-1으로 명명하였다. 균주 HX-1로부터의 자일라네이즈 생산을 증가시키는 배지 조건을 탐색하여 최적화된 TNX 배지(1% bacto tryptone, 0.7% 자일란, 1% NaCl; pH 7.0)에서 약 7.4배에 달하는 자일라네이즈 생산량의 증가가 가능하였다. 균주 HX-1이 분하는 자일라네이즈는 pH 7.0과 45oC에서 최적의 효소활성을 나타냈으며, beechwood 자일란을 기질로 하는 효소반응으로부터 xylobiose를 최종산물로 생산하는 endo-β-1,4-xylanase임을 확인하였다. 본 연구로부터 동정된 Paenibacillussp. HX-1은 다양한 산업에 응용이 가능한 새로운 자일라네이즈를 제공할 수 있는 중요한 균으로 사료된다.
Abstract
A xylanase-producing bacterium was isolated from a soil sample collected in Yongin city, Korea. The strain was aerobic and gram positive, and grew between pH 5.0 and 11.0, forming a yellow-colored colony. The strain was classified as a novel subspecies bacterium of Paenibacillus barcinonensis by 16S rRNA gene sequence similarity, phylogenetic analysis, phenotypic,and biochemical characteristics, and thus named Paenibacillus sp. HX-1. This strain produced extracellular endo-β-1,4-xylanase,and the best xylanolytic activity (205.17 unit/ml) was obtained at 96 h in an optimized TNX medium containing 1% (w/v)bacto tryptone, 1% (w/v) NaCl, and 0.7% (w/v) beechwood xylan at pH 7.0, 37oC and 200 rpm. The endo-β-1,4-xylanase produced by the strain HX-1 yielded xylobiose as the end product from beechwood xylan hydrolysis. The enzyme exhibited optimum pH and temperature at pH 7.0 and 45oC, respectively. The remarkable enhancing effect of the TNX medium on xylanase production by HX-1, in spite of its simple formula, may have great advantages for industrial applications of xylanase.
참고문헌 (28)
- Ali MK, "Characterization of thermostable Xyn10A enzyme from mesophilic Clostridium acetobutylicum ATCC 824", J. Ind. Microbiol. Biotechnol, 32, pp.12~18 (2005)
- Amaya-Delgado L, "Cloning and expression of a novel, moderately thermostable xylanase-encoding gene (Cfl xyn11A) from Cellulomonas flavigena", Bioresour. Technol, 101, pp.5539~5545 (2010)
- Beg QK, "Microbial xylanase and their industrial applications: A review", Appl. Microbiol. Biotechnol, 56, pp.326~338 (2001)
- 지원재, "Xylanase 생산균 Bacillus sp. MX47의 분리 및 동정", 한국미생물·생명공학회지, 40(4), pp.419~423 (2012), 한국미생물·생명공학회
- Chi W-J, "A novel alkaliphilic xylanase from the newly isolated mesophilic Bacillus sp. MX47: production, purification, and characterization", Appl. Biochem. Biotechnol, 168, pp.899~909 (2012)
- Chun JS, "Extaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences", Int. J. Syst. Evol. Microbiol, 57, pp.2259~2261 (2007)
- Felsenstein J., "PHYLIP (phylogeny inference package), v3.69. Distributed by the author", Department of Genome Sciences, pp.2009
- Hall TA., "BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95", 98, Nucleic Acids Symp. Ser, pp.1999, 41[DOI]
- HEO, SUNYEON, "Characterization of an Extracellular Xylanase in Paenibacillus sp. HY-8Isolated from an Herbivorous Longicorn Beetle", Journal of Microbiology and Biotechnology, 16(11), pp.1753~1759 (2006), 한국미생물·생명공학회
- Ito Y, "Cloning, expression, and cell surface localization of Paenibacillus sp. strain W-61 xylanase 5, a multidomain xylanase", Appl. Environ. Microbiol, 69, pp.6969~6978 (2003)
- Kim DY, "Novel GH10 xylanase, with a fibronectin type 3 domain, from Cellulosimicrobium sp. strain HY-13, a bacterium in the gut of Eisenia fetida", Appl. Environ. Microbiol, 75, pp.7275~7279 (2009)
- Kimura M., "The neutral theory of molecular evolution" (1983), Cambridge Univesity Press
- Ko CH, "Molecular cloning and characterization of a novel thermostable xylanase from Paenibacillus campinasensis BL11", Process Biochem, 45, pp.1638~1644 (2010)
- Komagata K, "Lipid and cell-wall analysis in bacterial systematic", Methods Microbiol, 19, pp.161~207 (1987)
- Mesbah M, "Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography", Int. J. Syst. Bacteriol, 39, pp.159~167 (1989)
- Miller GL, "Use of dinitrosalicylic acid reagent for determination of reducing sugar", Anal. Chem, 31, pp.426~428 (1959)
- Bacterial identification by gas chromatography of whole cell fatty acid, "Miller L", Hewlett-Packard, 228~241, pp.Hewlett-Packard Application note (1985)
- Ratanakhanokchai K, "Purification and properties of a xylan-binding endoxylanase from alkaliphilic Bacillus sp. strain K-1", Appl. Environ. Microbiol, 65, pp.694~697 (1999)
- Waeonukul Rattiya, "Cloning, Sequencing, and Expression of the Gene Encoding a Multidomain Endo-β-1,4-Xylanase from Paenibacillus curdlanolyticus B-6, and Characterization of the Recombinant Enzyme", Journal of Microbiology and Biotechnology, 19(3), pp.277~285 (2009), 한국미생물·생명공학회
- Sánchez MM, "Paenibacillus barcinonensis sp. nov., a xylanase-producing bacterium isolated from a rice field in the Ebro River delta", Int. J. Syst. Evol. Microbiol, 55, pp.935~939 (2005)
- Saitou N, "The neighbor-joining method: a new method for reconstructing phylogenetic trees", Mol. Biol. Evol, 4, pp.406~425 (1987)
- Identification of bacteria by gas chromatography of cellular fatty acids, "Sasser M.", MIDI Inc, ~, pp.MIDI Technical Note 101 (1997)
- Thompson JD, "CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice", Nucleic Acids Res, 22, pp.4673~4680 (1994)
- Valenzuela SV, "Recombinant expression of an alkali stable GH 10 xylanase from Paenibacillus barcinonensis", J. Agric. Food Chem, 58, pp.4814~4818 (2010)
- Watanabe S, "Cloning, expression and transglycosylation reaction of Paenibacillus sp. strain W-61 xylanase1", Biosci. Biotechnol. Biochem, 72, pp.951~958 (2008)
- Zhang J, "Thermostable recombinant xylanases from Nonomuraea flexuosa and Thermoascus aurantiacus show distinct properties in the hydrolysis of xylans and pretreated wheat straw", Biotechnol. Biofuels, 4, pp.12~25 (2011)
- Zhao, Yanyu, "Cloning, Expression, and Characterization of a New Xylanase from Alkalophilic Paenibacillus sp. 12-11", Journal of Microbiology and Biotechnology, 21(8), pp.861~868 (2011), 한국미생물·생명공학회
- Zheng, Hongchen, "Isolation, Purification, and Characterization of a Thermostable Xylanase from a Novel Strain, Paenibacillus campinasensis G1-1", Journal of Microbiology and Biotechnology, 22(7), pp.930~938 (2012), 한국미생물·생명공학회