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Cloning, expression and characterization of a novel cold-active and halophilic xylanase from Zunongwangia profunda

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Abstract

A new xylanase gene (xynA) from the marine microorganism Zunongwangia profunda was identified to encode 374 amino acid residues. Its product (XynA) showed the highest identity (42.78 %) with a xylanase from Bacillus sp. SN5 among the characterized xylanases. XynA exhibited the highest activity at pH 6.5 and 30 °C, retaining 23 and 38 % of the optimal activity at 0 and 5 °C, respectively. XynA was not only cold active, but also halophilic, and both its activity and thermostability could be significantly increased by NaCl, showing the highest activity (180 % of the activity) at 3 M NaCl and retaining nearly 100 % activity at 5 M NaCl, compared to the absence of NaCl. In the presence of 3 M NaCl, the k cat/K m value of XynA exhibited a 3.41-fold increase for beechwood xylan compared to no added NaCl, and the residual activity of XynA increased from 23 % (no added NaCl) to 58 % after 1 h incubation at 45 °C. This may be the first report concerning a cold-adapted xylanase from a non-halophilic species that displays the highest activity at a NaCl concentration range from 3 to 5 M. The features of cold activity and salt tolerance suggest the potential application of XynA in the food industry and bioethanol production from marine seaweeds.

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References

  • Ahmed S, Riaz S, Jamil A (2009) Molecular cloning of fungal xylanases: an overview. Appl Microbiol Biotechnol 84:19–35

    Article  CAS  PubMed  Google Scholar 

  • Bai W, Xue Y, Zhou C, Ma Y (2012) Cloning, expression and characterization of a novel salt-tolerant xylanase from Bacillus sp. SN5. Biotechnol Lett 34:2093–2099

    Article  CAS  PubMed  Google Scholar 

  • Bajpai P (1997) Microbial xylanolytic enzyme system: properties and applications. Adv Appl Microbiol 43:141–194

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Butt MS, Tahir-Nadeem M, Ahmad Z, Sultan MT (2008) Xylanases and their applications in baking industry. Food Technol Biotechnol 46:22–31

    CAS  Google Scholar 

  • Collins T, Gerday C, Feller G (2005) Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29:3–23

    Article  CAS  PubMed  Google Scholar 

  • Feller G, Gerday C (1997) Review Psychrophilic enzymes: molecular basis of cold adaptation. CMLS Cell Mol Life Sci 53:830–841

    Article  CAS  Google Scholar 

  • Fukuchi S, Yoshimune K, Wakayama M, Moriguchi M, Nishikawa K (2003) Unique amino acid composition of proteins in halophilic bacteria. J Mol Biol 327:347–357

    Article  CAS  PubMed  Google Scholar 

  • Fukumura M, Sakka K, Shimada K, Ohmiya K (1995) Nucleotide sequence of the Clostridium stercorarium xynB gene encoding an extremely thermostable xylanase, and characterization of the translated product. Biosci Biotech Bioch 59:40–46

    Article  CAS  Google Scholar 

  • Guo B, Chen XL, Sun CY, Zhou BC, Zhang YZ (2009) 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

    Article  CAS  PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • Hung KS, Liu SM, Fang TY, Tzou WS, Lin FP, Sun KH, Tang SJ (2011a) Characterization of a salt-tolerant xylanase from Thermoanaerobacterium saccharolyticum NTOU1. Biotechnol Lett 33:1441–1447

    Article  CAS  PubMed  Google Scholar 

  • Hung KS, Liu SM, Tzou WS, Lin FP, Pan CL, Fang TY, Sun KH, Tang SJ (2011b) Characterization of a novel GH10 thermostable, halophilic xylanase from the marine bacterium Thermoanaerobacterium saccharolyticum NTOU1. Process Biochem 46:1257–1263

    Article  CAS  Google Scholar 

  • Juturu V, Wu JC (2012) Microbial xylanases: engineering, production and industrial applications. Biotechnol Adv 30:1219–1227

    Article  CAS  PubMed  Google Scholar 

  • Khandeparker R, Verma P, Deobagkar D (2011) A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing. New Biotechnol 28:814–821

    Article  CAS  Google Scholar 

  • Khasin A, Alchanati I, Shoham Y (1993) Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6. Appl Environ Microbiol 59:1725–1730

    CAS  PubMed Central  PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  • Lee CC, Kibblewhite-Accinelli RE, Wagschal K, Robertson GH, Wong DW (2006a) Cloning and characterization of a cold-active xylanase enzyme from an environmental DNA library. Extremophiles 10:295–300

    Article  CAS  PubMed  Google Scholar 

  • Lee CC, Smith M, Kibblewhite-Accinelli RE, Williams TG, Wagschal K, Robertson GH, Wong DW (2006b) Isolation and characterization of a cold-active xylanase enzyme from Flavobacterium sp. Curr Microbiol 52:112–116

    Article  CAS  PubMed  Google Scholar 

  • Liu Z, Zhao X, Bai F (2012) Production of xylanase by an alkaline-tolerant marine-derived Streptomyces viridochromogenes strain and improvement by ribosome engineering. Appl Microbiol Biotechnol 97:4361–4368

    Article  PubMed  Google Scholar 

  • Menon G, Mody K, Keshri J, Jha B (2010) Isolation, purification, and characterization of haloalkaline xylanase from a marine Bacillus pumilus strain, GESF-1. Biotechnol Bioproc Eng 15:998–1005

    Article  CAS  Google Scholar 

  • Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  • Nystuen A (2000) Lasergene 99. Biotech Softw Internet Rep: Computer Softw J Sci 1:192–197

    Article  Google Scholar 

  • Oren A (2002) Diversity of halophilic microorganisms: environments, phylogeny, physiology, and applications. J Ind Microbiol Biotechnol 28:56–63

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Qin QL, Zhao DL, Wang J, Chen XL, Dang HY, Li TG, Zhang YZ, Gao PJ (2007) Wangia profunda gen. nov., sp. nov., a novel marine bacterium of the family Flavobacteriaceae isolated from southern Okinawa Trough deep-sea sediment. FEMS Microbiol Lett 271:53–58

    Article  CAS  PubMed  Google Scholar 

  • Qin QL, Zhang XY, Wang XM, Liu GM, Chen XL, Xie BB, Dang HY, Zhou BC, Yu J, Zhang YZ (2010) The complete genome of Zunongwangia profunda SM-A87 reveals its adaptation to the deep-sea environment and ecological role in sedimentary organic nitrogen degradation. BMC Genom 11:247

    Article  Google Scholar 

  • Russell NJ (2000) Toward a molecular understanding of cold activity of enzymes from psychrophiles. Extremophiles 4:83–90

    Article  CAS  PubMed  Google Scholar 

  • Setati ME (2010) Diversity and industrial potential of hydrolase-producing halophilic/halotolerant eubacteria. Afr J Biotechnol 9:1555–1560

    CAS  Google Scholar 

  • Sheridan PP, Panasik N, Coombs JM, Brenchley JE (2000) Approaches for deciphering the structural basis of low temperature enzyme activity. BBA Protein Struct Mol 1543:417–433

    Article  CAS  Google Scholar 

  • Sunna A, Bergquist PL (2003) A gene encoding a novel extremely thermostable 1,4-β-xylanase isolated directly from an environmental DNA sample. Extremophiles 7:63–70

    CAS  PubMed  Google Scholar 

  • Trincone A (2011) Marine biocatalysts: enzymatic features and applications. Mar Drugs 9:478–499

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Van Den Burg B (2003) Extremophiles as a source for novel enzymes. Curr Opin Microbiol 6:213–218

    Article  PubMed  Google Scholar 

  • Wang G, Luo H, Wang Y, Huang H, Shi P, Yang P, Meng K, Bai Y, Yao B (2011) A novel cold-active xylanase gene from the environmental DNA of goat rumen contents: direct cloning, expression and enzyme characterization. Bioresour Technol 102:3330–3336

    Article  CAS  PubMed  Google Scholar 

  • Wang SY, Hu W, Lin XY, Wu ZH, Li YZ (2012) A novel cold-active xylanase from the cellulolytic myxobacterium Sorangium cellulosum So9733-1: gene cloning, expression, and enzymatic characterization. Appl Microbiol Biotechnol 93:1503–1512

    Article  CAS  PubMed  Google Scholar 

  • Ward OP, Moo-Young M, Venkat K (1989) Enzymatic degradation of cell wall and related plant polysaccharides. Crit Rev Biotechnol 8:237–274

    Article  CAS  PubMed  Google Scholar 

  • Winterhalter C, Liebl W (1995) Two Extremely Thermostable Xylanases of the Hyperthermophilic Bacterium Thermotoga maritima MSB8. Appl Environ Microbiol 61:1810–1815

    CAS  PubMed Central  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Zhang F, Hu SN, Chen JJ, Lin LB, Wei YL, Tang SK, Xu LH, Li WJ (2012) Purification and partial characterisation of a thermostable xylanase from salt-tolerant Thermobifida halotolerans YIM 90462 T. Process Biochem 47:225–228

    Article  CAS  Google Scholar 

  • Zhou J, Gao Y, Dong Y, Tang X, Li J, Xu B, Mu Y, Wu Q, Huang Z (2012) A novel xylanase with tolerance to ethanol, salt, protease, SDS, heat, and alkali from actinomycete Lechevalieria sp. HJ3. J Ind Microbiol Biotechnol 39:965–975

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by grants from the National Natural Science Foundation of China (No.J1103510).

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Authors and Affiliations

  1. State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China

    Xiaoshuang Liu, Zongqing Huang, Xiangnan Zhang & Ziduo Liu

  2. Key Laboratory of Marine Biogenetic Resources, The Third Institute of Oceanography, State of Oceanic Administration, Xiamen, 361005, People’s Republic of China

    Zongze Shao

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  1. Xiaoshuang Liu
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  2. Zongqing Huang
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  3. Xiangnan Zhang
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  4. Zongze Shao
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  5. Ziduo Liu
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Correspondence to Ziduo Liu.

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Communicated by F. Robb.

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Liu, X., Huang, Z., Zhang, X. et al. Cloning, expression and characterization of a novel cold-active and halophilic xylanase from Zunongwangia profunda . Extremophiles 18, 441–450 (2014). https://doi.org/10.1007/s00792-014-0629-x

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