Abstract
The novel fungus Aspergillus niveus RS2 isolated from rice straw showed relatively high xylanase production after 5 days of fermentation. Of the different xylan-containing agricultural by-products tested, rice husk was the best substrate; however, maximum xylanase production occurred when the organism was cultured on purified xylan. Yeast extract was found to be the best nitrogen source for xylanase production, followed by ammonium sulfate and peptone. The optimum pH for maximum enzyme production was 8 (18.2 U/ml); however, an appreciable level of activity was obtained at pH 7 (10.9 U/ml). Temperature and pH optima for xylanase were 50°C and 7.0, respectively; however the enzyme retained considerably high activity under high temperature (12.1 U/ml at 60°C) and high alkaline conditions (17.2 U/ml at pH 8 and 13.9 U/ml at pH 9). The enzyme was strongly inhibited by Hg2+, while Mn2+ was slight activator. The half-life of the enzyme was 48 min at 50°C. The enzyme was purified by 5.08-fold using carboxymethyl-sephadex chromatography. Zymogram analysis suggested the presence of a single candidate xylanase in the purified preparation. SDS-PAGE revealed a molecular weight of approximately 22.5 kDa. The enzyme had K m and V max values of 2.5 and 26 μmol/mg per minute, respectively.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Badhan AK, Chadha BS, Sonia KG, Saini HS, Bhat MK (2004) Functionally diverse multiple xylanases of thermophilic fungus Myceliophthora sp. IMI−387099. Enzyme Microb Technol 35:460–466
Bakir U, Yavascoglu S, Guvence F, Erroyin A (2001) An endo- β-1,4-xylanase from Rhizopus oryzae: production, partial purification and biochemical characterization. Enzyme Microb Technol 29:328–334
Bakri Y, Jacques P, Thonart P (2003) Xylanase production by Penicillium canescens 10–10c in solid state fermentation. Appl Biochem Biotechnol 108:737–748
Beg QA, Kapoor M, Mahajan G, Hoondal S (2001) Microbial xylanases and their industrial applications: a review. Appl Microbiol Biotechnol 56:326–338
Belancic A, Scarpa J, Peirano A, Diaz R, Steiner J, Eyzayuirre J (1995) Penicillium purpurogenum produces several xylanases: purification and properties of two of the enzymes. J Biotechnol 41:71–79
Carmona EC, Fialho MB, Buchgnani EB, Coelho GD, Brocheto-Braga MR, Jorge JA (2005) Production, purification and characterization of a minor form of xylanase from Aspergillus versicolor. Process Biochem 40:359–364
Cesar T, Mrsa V (1996) Purification and properties of xylanase produced by Thermomyces lanuginosus. Enzyme Microb Technol 19:289–296
Fialho MB, Carmona EC (2004) Purification and characterization of xylanases from Aspergillus giganteus. Folia Microbiol 49:13–18
Gilbert HJ, Hazlewood GP (1993) Bacterial cellulases and xylanases. J Gen Microbiol 139:187–194
Haltrich D, Nidetzky B, Kulbe K, Steiner W, Zupancic S (1996) Production of fungal xylanases. Bioresour Technol 58:137–161
Jiang Z, Deng W, Li L, Ding C, Kusakabe I, Tan S (2004) A novel, ultra-large xylanolytic complex (xylanosome) secreted by Streptomyces olivaceoviridis. Biotechnol Lett 26:431–436
Kango N, Agrawal SC, Jain PC (2003) Production of xylanase by Emericella nidulans NK-62 on low value lignocellulosic substrates. World J Microbiol Biotechnol 19:691–694
Kuhad RC, Singh A (1993) Lignocellulosic Biotechnology: current and future prospects. Crit Rev Biotechnol 13:151–172
Lin J, Ndlovu LM, Singh S, Pillay B (1999) Purification and bio-chemical characteristics of β-d-xylanase from a thermophilic fungus Thermomyces lanuginosus-SSBP. Biotechnol Appl Biochem 30:73–79
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Miller GL (1959) Use of dinitrosalicylic acid for the determination of reducing sugar. Ann Chem 31:426– 428
Purkarthofer H, Sinner M, Steiner W (1993) Cellulase-free xylanase from Thermomyces lanuginosus: optimization of production in submerged and solid state culture. Enzyme Microb Technol 15:677–682
Raper KB, Fennell DI (1965) The genus Aspergillus. Williams & Wilkins, Baltimore
Ruckmanl A, Rajendran A (2001) Production of cellulase poor xylanase by an alkali tolerant strain of Aspergillus flavus. Indian J Microbiol 41:115–118
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Shah AR, Madamwar D (2005) Xylanase production by a newly isolated Aspergillus foetidus strain and its characterization. Process Biochem 40:1763–1771
Sharma P, Bajaj BK (2005) Production and partial characterization of alkali tolerant xylanase from an alkalophilic Streptomyces sp. CD3. J Sci Industrial Res 64:688–698
Singh S, Madlala AM, Prior BA (2003) Thermomyces lanuginosus: properties of strains and their hemicellulases. FEMS Microbiol Rev 27:3–16
Srinivasan MC, Rele MV (1995) Cellulase-free xylanase from microorganisms and their applications in pulp and paper biotechnology: an overview. Indian J Microbiol 35:93–101
Subramanian S, Prema P (2000) Cellulase-free xylanases from Bacillus and other microorganisms. FEMS Microbiol Lett 183:1–7
Xiong H, Weymarn N, Leisola M, Turunen O (2004) Influence of pH on the production of xylanase by Trichoderma reesei C-30. Process Biochem 39:731–736
Acknowledgements
The authors wish to thank the Head, Department of Biotechnology, University of Jammu, Jammu, for the laboratory facilities, and Dr. Yash Pal Reader, Department of Botany, University of Jammu, Jammu, for kindly helping in the identification of the fungus.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sudan, R., Bajaj, B.K. Production and biochemical characterization of xylanase from an alkalitolerant novel species Aspergillus niveus RS2. World J Microbiol Biotechnol 23, 491–500 (2007). https://doi.org/10.1007/s11274-006-9251-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11274-006-9251-0