Purification and characterization of endoxylanase Xln-2 from Aspergillus niger B03
An extracellular multiple form of endoxylanase was isolated from the xylanolytic complex of Aspergillus niger B03. The enzyme was purified to a homogenous form using ultrafiltration, anion exchange chromatography, and gel filtration. It was a nonglycosylated protein with a molecular weight of 20,000 Da as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 21,000 Da as determined by gel filtration. The optimal pH for the enzyme action was 5.0 and the optimal temperature was 55 °C. Endoxylanase stability was significantly improved in the presence of glycerol and sorbitol. The enzyme activity was activated by Mn2+ and Co2+, and it was inhibited by Ag+, Cu2+, Fe3+, Fe2+, and Pb2+. The substrate specificity and the product profile of the enzyme suggested that it was an endoxylanase. The enzyme showed a synergism with another endoxylanase from Aspergillus niger B03 in xylan hydrolysis.
Purification and characterization of endoxylanase Xln-2 from Aspergillus niger B03
An extracellular multiple form of endoxylanase was isolated from the xylanolytic complex of Aspergillus niger B03. The enzyme was purified to a homogenous form using ultrafiltration, anion exchange chromatography, and gel filtration. It was a nonglycosylated protein with a molecular weight of 20,000 Da as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 21,000 Da as determined by gel filtration. The optimal pH for the enzyme action was 5.0 and the optimal temperature was 55 °C. Endoxylanase stability was significantly improved in the presence of glycerol and sorbitol. The enzyme activity was activated by Mn2+ and Co2+, and it was inhibited by Ag+, Cu2+, Fe3+, Fe2+, and Pb2+. The substrate specificity and the product profile of the enzyme suggested that it was an endoxylanase. The enzyme showed a synergism with another endoxylanase from Aspergillus niger B03 in xylan hydrolysis.
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- Sandrim VC, Rizzatti AC, Terenzi HF et al. Purifi cation and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching. Process Biochem 40: 1823-1828, 2005.
- Kolenová K, Vršanská M, Biely P. Purifi cation and characterization of two minor endo-β-1,4-xylanases of Schizophyllum commune. Enzyme Microbial Technol 36: 903- 910, 2005.
- Fawzi EM. Highly thermostable purifi ed xylanase from Rhizomucor miehei NRRL 3169. Ann Microbiol 60: 363-368, 2010.
- Beg QK, Kapoor K, Mahajan L et al. Microbial xylanases and their industrial applications: a review. Appl Microbiol Biotechnol 56: 326-338, 2001. 5. Collins T, Gerday C, Feller G. Xylanases, xylanase families and extremophilic xylanases. FEMS Microbiol Rev 29: 3-23, 2005.
- Li XL, Ljungdahl LG. Cloning, sequencing, and regulation of a xylanase gene from the fungus Aureobasidium pullulans Y-2311-1. Appl Environ Microbiol 60: 3160-3169, 1994.
- Th omson JA. Molecular biology of xylan degradation. FEMS Microbiol Rev 104: 65-82, 1993. 8. Biely P, Vrsanská M, Tenkanen M et al. Endo-β-1,4-xylanase families: diff erences in catalytic properties. J Biotechnol 57: 151-166, 1997.
- Dobrev G, Zhekova B, Delcheva G et al. Purifi cation and characterization of endoxylanase Xln-1 from Aspergillus niger B03. World J Microbiol Biotechnol 25: 2095-2102, 2009.
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680- 685, 1970.
- Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31: 426-428, 1959.
- Ronen R, Zauberman G, Akerman M et al. Xylanase and xylosidase activities in avocado fruit. Plant Physiol 95: 961- 964, 1991.
- Ponpium P, Ratanakhanokchai K, Kyu K. Isolation and properties of a cellulosome-type multienzyme complex of the thermophilic Bacteroides sp. strain P-1. Enzyme Microbial Technol 26: 459-465, 2000.
- Delcheva G, Pishtiyski I, Dobrev G et al. Immobilization of Aspergillus niger pectinase on polyacrylonitrile copolymer membrane. Trends Appl Sci Res 2: 419-425, 2007.
- Bailey MJ, Biely P, Poutanen K. Interlaboratory testing of method for assay of xylanase activity. J Biotechnol 23: 257-270, 1992.
- Dubois M, Gilles KA, Hamilton JK et al. Colorimetric method for determination of sugars and related substances. Anal Chem 28: 350-356, 1956.
- Lowry OH, Rosebrough NJ, Farr AL et al. Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265-275, 1951.
- Lemos JLS, Bon EPS, Santana MFE et al. Th ermal stability of xylanases produced by Aspergillus awamori. Braz J Microbiol 31: 206-211, 2000.
- Lv Z, Yang J, Yuan H. Production, purifi cation and characterization of an alkaliphilic endo-β-1,4-xylanase from a microbial community EMSD5. Enzyme Microbial Technol 43: 343-348, 2008.
- Maalej I, Belhaj I, Masmoudi NF et al. Highly thermostable xylanase of the thermophilic fungus Talaromyces thermophilus: purifi cation and characterization. Appl Biochem Biotechnol 158: 200-212, 2009.
- Martínez-Trujillo A, Pérez-Avalos O, Ponce-Noyola T. Enzymatic properties of a purifi ed xylanase from mutant PN- 120 of Cellulomonas fl avigena. Enzyme Microbial Technol 32: 401-406, 2003.