Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Cloning, expression, and characterization of a new xylanase with broad temperature adaptability from Streptomyces sp. S9

Abstract

A new xylanase gene, xynAS9, was cloned from Streptomyces sp. S9, which was isolated from Turpan Basin, China. The full-length gene consists of 1,395 bp and encodes 465 amino acids including 38 residues of a putative signal peptide. The overall amino acid sequence shares the highest identity (50.8%) with a putative endo-1,4-β-xylanase from Streptomyces avermitilis of the glycoside hydrolase family 10. The gene fragment encoding the mature xylanase was expressed in Escherichia coli BL21 (DE3). The recombinant protein was purified to electrophoretic homogeneity and subsequently characterized. The optimal pH and temperature for the recombinant enzyme were 6.5 and 60°C, respectively. The enzyme showed broad temperature adaptability, retaining more than 65% of the maximum activity when assayed at 50–80°C. The enzyme also had good thermal and pH stability. The K m values for oat spelt xylan and birchwood xylan substrates were 2.85 and 2.43 mg ml−1, with the V max values of 772.20 and 490.87 μmol min−1 mg−1, respectively. The hydrolysis products of xylan were mainly xylose and xylobiose. These favorable properties should make XynAS9 a good candidate in various industrial applications.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

References

  1. Ali MK, Fukumura M, Sakano K, Karita S, Kimura T, Sakka K, Ohmiya K (1999) Cloning, sequencing, and expression of the gene encoding the Clostridium stercorarium xylanase C in Escherichia coli. Biosci Biotechnol Biochem 63:1596–1604

  2. Ali MK, Rudolph FB, Bennett GN (2004) Thermostable xylanase10B from Clostridium acetobutylicum ATCC824. J Ind Microbiol Biotechnol 31:229–234

  3. Biely P (1985) Microbial xylanolytic systems. Trends Biotechnol 3:286–290

  4. 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

  5. Bottoli AP, Kertesz-Chaloupková K, Boulianne RP, Granado JD, Aebi M, Kües U (1999) Rapid isolation of genes from an indexed genomic library of C. cinereus in a novel pab1 + cosmid. J Microbiol Methods 35:129–141

  6. 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

  7. Cardoso OAV, Filho EXF (2003) Purification and characterization of a novel cellulase-free xylanase from Acrophialophora nainiana. FEMS Microbiol Lett 223:309–314

  8. Chantasingh D, Pootanakit K, Champreda V, Kanokratana P, Eurwilaichitr L (2006) Cloning, expression, and characterization of a xylanase 10 from Aspergillus terreus (BCC129) in Pichia pastoris. Protein Expr Purif 46:143–149

  9. Collins T, Meuwis MA, Stals I, Claeyssens M, Feller G, Gerday C (2002) A novel family 8 xylanase, functional and physicochemical characterization. J Biol Chem 277:35133–35139

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

  11. Decelle B, Tsang A, Storms RK (2004) Cloning, functional expression and characterization of three Phanerochaete chrysosporium endo-β-1,4-xylanases. Curr Genet 46:166–175

  12. Dutta T, Sengupta R, Sahoo R, Sinha Ray S, Bhattacharjee A, Ghosh S (2007) A novel cellulase free alkaliphilic xylanase from alkali tolerant Penicillium citrinum: production, purification and characterization. Lett Appl Microbiol 44:206–211

  13. Fujimoto Z, Kuno A, Kaneko S, Yoshida S, Kobayashi H, Kusakabe I, Mizuno H (2000) Crystal structure of Streptomyces olivaceoviridis E-86 β-xylanase containing xylan-binding domain. J Mol Biol 300:575–585

  14. Gessesse A (1998) Purification and properties of two thermostable alkaline xylanases from an alkaliphilic Bacillus sp. Appl Environ Microbiol 64:3533–3535

  15. Grüninger H, Fiechter A (1986) A novel, highly thermostable d-xylanase. Enzyme Microb Technol 8:309–314

  16. Heck JX, Flôres SH, Hertz PF, Ayub MA (2006a) Statistical optimization of thermo-tolerant xylanase activity from Amazon isolated Bacillus circulans on solid-state cultivation. Bioresour Technol 97:1902–1906

  17. Heck JX, de Barros Soares LH, Hertz PF, Ayub MA (2006b) Purification and properties of a xylanase produced by Bacillus circulans BL53 on solid-state cultivation. Biochem Eng J 32:179–184

  18. Huo B, Pan HX, Feng L, Wan ZF, Shi YH (1996) Study on the thermaturic neutral proteinase of the thermophilic Bacillus XJT9503. Biotechnol 6:24–27 (in Chinese)

  19. Kimura T, Ito J, Kawano A, Makino T, Kondo H, Karita S, Sakka K, Ohmiya K (2000) Purification, characterization, and molecular cloning of acidophilic xylanase from Penicillium sp.40. Biosci Biotechnol Biochem 64:1230–1237

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

  21. Li N, Yang PL, Wang YR, Luo HY, Meng K, Wu NF, Fan YL, Yao B (2008) Cloning, expression, and characterization of protease-resistant xylanase from Streptomyces fradiae var. k11. J Microbiol Biotechnol 18:410–416

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

  23. Moreau A, Roberge M, Manin C, Shareck F, Kluepfel D, Morosoli R (1994) Identification of two acidic residues involved in the catalysis of xylanase A from Streptomyces lividans. Biochem J 302:291–295

  24. 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

  25. Ratanachomsri U, Sriprang R, Sornlek W, Buaban B, Champreda V, Tanapongpipat S, Eurwilaichitr L (2006) Thermostable xylanase from Marasmius sp.: purification and characterization. J Biochem Mol Biol 39:105–110

  26. 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

  27. Sá-Pereira P, Mesquita A, Duarte JC, Barros MRA, Costa-Ferreira M (2002) Rapid production of thermostable cellulase-free xylanase by a strain of Bacillus subtilis and its properties. Enzyme Microb Technol 30:924–933

  28. Simpson HD, Haufler UR, Daniel RM (1991) An extremely thermostable xylanase from the thermophilic eubacterium Thermotoga. Biochem J 277:413–417

  29. Subramaniyan S, Prema P (2002) Biotechnology of microbial xylanases: enzymology, molecular biology, and application. Crit Rev Biotechnol 22:33–64

  30. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

  31. Tsujibo H, Miyamoto K, Kuda T, Minami K, Sakamoto T, Hasegawa T, Inamori Y (1992) Purification, properties, and partial amino acid sequences of thermostable xylanases from Streptomyces thermoviolaceus OPC-520. Appl Environ Microbiol 58:371–375

  32. Wood PJ, Erfle JD, Teather RM (1988) Use of complex formation between Congo red and polysaccharide in detection and assay of polysaccharide hydrolases. Methods Enzymol 160:59–74

  33. Wu SJ, Liu B, Zhang XB (2006) Characterization of a recombinant thermostable xylanase from deep-sea thermophilic Geobacillus sp. MT-1 in East Pacific. Appl Microbiol Biotechnol 72:1210–1216

  34. Yang PL, Shi PJ, Wang YR, Bai YG, Meng K, Luo HY, Yuan TZ, Yao B (2007) Cloning and overexpression of a Paenibacillus β-glucanase in Pichia pastoris: purification and characterization of the recombinant enzyme. J Microbiol Biotechnol 17:58–66

  35. Yi X, Xie ZJ, Deng AH, Wang N, Erkin R (2006) Isolation and identification of two xylanase-producing extremely alkali-tolerant strains of Bacillus halodurans from Turpan in China. Acta Microbiologica Sinica 46:951–955 (in Chinese)

  36. Zhang GM, Huang J, Huang GR, Ma LX, Zhang XE (2007) Molecular cloning and heterologous expression of a new xylanase gene from Plectosphaerella cucumerina. Appl Microbiol Biotechnol 74:339–346

  37. Zhang HL, Yao B, Yuan TZ, Wang YR, Cao SS, Fan YL (2002) Purification and properties of 43 kD xylanase XYNA from Streptomyces olivaceoviridis A1. J Agric Biotechnol 10:10–14 (in Chinese)

Download references

Acknowledgments

This work was supported by National High Technology Research and Development Program of China (863 Program; no. 2007AA100601) and National Key Technology R&D Program of China (no. 2006BAD12B05-03).

Author information

Correspondence to Bin Yao.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Li, N., Meng, K., Wang, Y. et al. Cloning, expression, and characterization of a new xylanase with broad temperature adaptability from Streptomyces sp. S9. Appl Microbiol Biotechnol 80, 231 (2008). https://doi.org/10.1007/s00253-008-1533-z

Download citation

Keywords

  • Streptomyces sp. S9
  • Turpan Basin
  • Xylanase
  • Xylose