Abstract
Although a lot of xylanases are studied, only a few xylanases from marine microorganisms have been reported. A new xylanase gene, xynA, was cloned from marine bacterium Glaciecola mesophila KMM 241. Gene xynA contains 1,272 bp and encodes a 423-amino acid xylanase precursor. The recombinant xylanase, XynA, expressed in Escherichia coli BL21 is a monomer with a molecular mass of 43 kDa. Among the characterized xylanases, XynA shares the highest identity (46%) to the xylanase from Flavobacterium sp. strain MSY2. The optimum pH and temperature for XynA is 7.0 and 30 °C. XynA retains 23% activity and 27% catalytic efficiency at 4 °C. XynA has low thermostability, remaining 20% activity after 60-min incubation at 30 °C. Its apparent melting temperature (T m) is 44.5 °C. These results indicate that XynA is a cold-active xylanase. XynA shows a high level of salt-tolerance, with the highest activity at 0.5 M NaCl and retaining 90% activity in 2.5 M NaCl. It may be the first salt-tolerant xylanase reported. XynA is a strict endo-β-1,4-xylanase with a demand of at least four sugar moieties for effective cleavage. It efficiently hydrolyzes xylo-oligosaccharides and xylan into xylobiose and xylotriose without producing xylose, suggesting its potential in xylo-oligosaccharides production.
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References
Akila G, Chandra TS (2003) A novel cold-tolerant Clostridium strain PXYL1 isolated from a psychrophilic cattle manure digester that secretes thermolabile xylanase and cellulase. FEMS Microbiol Lett 219:63–67
Ali MK, Rudolph FB, Bennett GN (2004) Thermostable xylanase 10B from Clostridium acetobutylicum ATCC824. J Ind Microbiol Biotechnol 31:229–234
Araki T, Tani S, Maeda K, Hashikawa S, Nakagawa H, Morishita T (1999) Purification and characterization of β-1, 3-xylanase from a marine bacterium, Vibrio sp. XY-214. Biosci Biotechnol Biochem 63:2017–2019
Badal CS (2002) Production, purification and properties of xylanase from a newly isolated Fusarium proliferatum. Process Biochem 37:1279–1284
Biely P, Vrsanská M, Tenkanen M, Kluepfel D (1997) Endo-β-1, 4-xylanase families: differences in catalytic properties. J Biotechnol 57:151–166
Blanco J, Coque JJR, Velasco J, Martín JF (1997) Cloning, expression in Streptomyces lividans and biochemical characterization of a thermostable endo-β-1, 4-xylanase of Thermomonospora alba ULJB1 with cellulose-binding ability. Appl Microbiol Biotechnol 48:208–217
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
Bradner JR, Sidhu RK, Gillings M, Nevalainen KM (1999) Hemicellulase activity of antarctic microfungi. J Appl Microbiol 87:366–370
Collins T, Meuwis MA, Stals I, Claeyssens M, Feller G, Gerday C (2002) A novel family 8 xylanase: functional and physico-chemical characterization. J Biol Chem 277:35133–35139
Collins T, Gerday C, Feller G (2005) Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29:3–23
Collins T, Hoyoux A, Dutron A, Georis J, Genot B, Dauvrin T, Arnaut F, Gerday C, Feller G (2006) Use of glycoside hydrolase family 8 xylanases in baking. J Cereal Sci 43:79–84
Fialho MB, Carmona EC (2004) Purification and characterization of xylanase from Aspergillus giganteus. Folia Microbiol 49:13–18
Gilkes NR, Henrissat B, Kilburn DG, Miller RC Jr, Warren RA (1991) Domains in microbial 4-glycanases: sequence conservation, function, and enzyme families. Microbiol Rev 55:303–315
Gruppen H, Hamer RJ, Voragen AGJ (1992) Water-unextractable cell wall material from wheat flour. 2. Fractionation of alkali-extracted polymers and comparison with waterextractable arabinoxylans. J Cereal Sci 16:53–67
Gupta S, Bhushan B, Hoondal GS (2000) Isolation, purification and characterization of xylanase from Staphylococcus sp. SG-13 and its application in biobleaching of kraft pulp. J Appl Microbiol 88:325–334
Henrissat B (1991) A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 280:309–316
Hou YH, Wang TH, Long H, Zhu HY (2006) Novel cold-adaptive Penicillium strain FS010 secreting thermo-labile xylanase isolated from Yellow Sea. Acta Biochim Biophys Sin 38(2):142–149
Hu Y, Zhang G, Li A, Chen J, Ma L (2008) Cloning and enzymatic characterization of a xylanase gene from a soil-derived metagenomic library with an efficient approach. Appl Microbiol Biotechnol 80:823–830
Humphry DR, George A, Black GW, Cummings SP (2001) Flavobacterium frigidarium sp. nov., an aerobic, psychrophilic, xylanolytic and laminarinolytic bacterium from Antarctica. Int J Syst Evol Microbiol 51:1235–1243
Inglis GD, Popp AP, Selinger LB, Kawchuk LM, Gaudet DA, McAllister TA (2000) Production of cellulases and xylanases by low-temperature basidiomycetes. Can J Microbiol 46:860–865
Irena R, Jacek P, Stanislaw B (2006) Isolation and properties of Aspergillus niger IBT-90 xylanase for bakery. Appl Microbiol Biotechnol 69:665–671
Kormelink FJM, Voragen AG (1993) Degradation of different [(glucurono) arabino] xylans by a combination of purified xylan-degrading enzymes. Appl Microbiol Biotechnol 38:688–695
Krisana A, Rutchadaporn S, Jarupan G, Lily E, Sutipa T, Kanyawim K (2005) Endo-1, 4-β-xylanase B from Aspergillus cf. niger BCC14405 isolated in Thailand: purification, characterization and gene isolation. J Biochem Mol Biol 38:17–23
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lee CC, Smith M, Kibblewhite-Accinelli RE, Williams TG, Wagschal K, Robertson GH, Wong DW (2006a) Isolation and characterization of a cold-active xylanase enzyme from Flavobacterium sp. Curr Microbiol 52:112–116
Lee CC, Kibblewhite-Accinelli RE, Wagschal K, Robertson GH, Wong DW (2006b) Cloning and characterization of a cold-active xylanase enzyme from an environmental DNA library. Extremophiles 10:295–300
Li N, Meng K, Wang Y, Shi P, Luo H, Bai Y, Yang P, Yao B (2008) Cloning, expression, and characterization of a new xylanase with broad temperature adaptability from Streptomyces sp. S9. Appl Microbiol Biotechnol 80:231–240
Liu R, Qu Y, Jiang Y, Gao P (1999) Purification and characterization of alkaline xylanases from Pseudomonas G6–2. Wei Sheng Wu Xue Bao 39:132–136
Miller GL, Blum R, Glennon WE, Burton AL (1960) Measurement of carboxymethyl cellulase activity. Anal Biochem 2:127–132
Morosoli R, Bertrand JL, Mondou F, Shareck F, Kluepfel D (1986) Purification and properties of a xylanase from Streptomyces lividans. Biochem J 239:587–592
Petrescu I, Lamotte-Brasseur J, Chessa JP, Ntarima P, Claeyssens M, Devreese B, Marino G, Gerday C (2000) Xylanase from the psychrophilic yeast Cryptococcus adeliae. Extremophiles 4:137–144
Romanenko LA, Zhukova NV, Rohde M, Lysenko AM, Mikhailov VV, Stackebrandt E (2003) Glaciecola mesophila sp. nov., a novel marine agar-digesting bacterium. Int J Syst Evol Microbiol 53:647–651
Ruiz-Arribas A, Fernández-Abalos JM, Sánchez P, Garda AL, Santamariá RI (1995) Overproduction, purification, and biochemical characterization of a xylanase (Xys1) from Streptomyces halstedii JM8. Appl Environ Microbiol 61:2414–2419
Saito H, Miura K (1963) Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochim Biophys Acta 72:619–629
Turkiewiz M, Kalinowska H, Zielinska M, Bielecki S (2000) Purification and characterisation of two endo-1, 4-xylanases from Antarctic krill, Euphasia superba Dana. Comp Biol Physiol Part B 127:325–335
Wu S, Liu B, Zhang X (2006) Characterization of a recombinant thermostable xylanase from deep-sea thermophilic Geobacillus sp. MT-1 in East Pacific. Appl Microbiol Biotechnol 72:1210–1216
Yamaura I, Koga T, Matsumoto T, Kato T (1997) Purification and some properties of endo-1, 4-β-D-xylanase from a fresh-water mollusc, Pomacea insularus (de Ordigny). Biosci Biotechnol Biochem 61:615–620
Yuan KP, Vrijmoed LL, Feng MG (2005) Survey of coastal mangrove fungi for xylanase production and optimized culture and assay conditions. Wei Sheng Wu Xue Bao 45(1):91–96
Zhang G, Huang J, Huang G, Ma L, Zhang X (2007) Molecular cloning and heterologous expression of a new xylanase gene from Plectosphaerella cucumerina. Appl Microbiol Biotechnol 74:339–346
Acknowledgment
The work was supported by Hi-Tech Research and Development program of China (2007AA091504, 2007AA091903), COMRA Program (DYXM-115-02-2-6), and Specialized Research Fund for the Doctoral Program of Higher Education (20060422053).
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Guo, B., Chen, XL., Sun, CY. et al. Gene cloning, expression and characterization of a new cold-active and salt-tolerant endo-β-1,4-xylanase from marine Glaciecola mesophila KMM 241. Appl Microbiol Biotechnol 84, 1107–1115 (2009). https://doi.org/10.1007/s00253-009-2056-y
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DOI: https://doi.org/10.1007/s00253-009-2056-y