The most promising application of xylanases (E.C. 3.2.1.8) is in the prebleaching of kraft pulp. The present paper reports bleaching effects of a thermoalkali stable xylanase from Cellulosimicrobium sp. MTCC 10645. The bacterium produced thermo-alkali stable xylanase in a basal medium supplemented with wheat bran (1% w/v), which was optimally active at pH 7.0 and 50°C. The xylanase was stable at temperature 50°C for 1?h and retained up to 86% of the activity. The xylanase was stable in a broad pH range of 6.0–11.0 for 1?h at 50°C. Metal ions Ca+2, Hg+2, and Pb+2 were inhibitory for xylanase retaining 72.3%, 35.07% and 36.7% relative activity at 10?mM concentration, whereas Fe+2, Cu+2, Mn+2, Na+2, Co+2, and Zn+2 were inducers at concentrations of 5?mM and 10?mM. The enzyme exhibited greater binding affinity exclusively for xylans but not for avicel, CMC, cellobiose, starch, or p-nitrophenyl xylopyranoside. Parachloromercuric benzoate and iodoacetamide were found stimulatory, while potassium permanganate, cysteine, and cystine markedly reduced the activity. The xylanase dose of 2.0?U/g dry weight pulp of 10% consistency gave optimum bleach boosting of kraft pulp at pH 8.0 and temperature 50°C for 5?h reaction time. 1. Introduction The most promising application of xylanases (E.C. 3.2.1.8) is in the prebleaching of kraft pulp. The pulp and paper industry is modifying its pulping, bleaching, and effluent treatment technologies to reduce the environmental impact of mill effluents. Prebleaching of kraft pulps with xylanases lowers chlorine charges, which reduce chloroorganic discharges [1–3]. Tremblay and Archibald reported the delignification of unbleached softwood and hardwood kraft pulps [4]. Thus reducing the Cl2 required to achieve a given degree of bleaching [3, 5–7]. The public concern on the impact of pollutants from paper and pulp industries, which use chlorine as the bleaching agent act as strong driving force in developing biotechnology aided techniques for novel bleaching that is biobleaching [8, 9]. The occurrence of cellulase contamination is posing a major threat in applying the xylanases in biobleaching. The cellulases easily result in the hydrolysis of cellulose, which should be the main recovered product in paper industry. However, the enzyme preparations from microorganisms producing higher levels of xylanases with no cellulase activity can be applied in paper industry because the loss of pulp viscosity is at minimum level [10]. Xylanases have been reported from bacteria, fungi, actinomycetes, and yeasts [11–14]. The use of abundantly
References
[1]
R. Koponer, “Enzyme systems prove their potential,” Pulp and Paper International, vol. 33, no. 11, pp. 20–25, 1991.
[2]
N. Kulkarni and M. Rao, “Application of xylanase from alkaliphilic thermophilic Bacillus sp. NCIM 59 in biobleaching of bagasse pulp,” Journal of Biotechnology, vol. 51, no. 2, pp. 167–173, 1996.
[3]
L. Viikari, A. Kantelinen, J. Sundquist, and M. Linko, “Xylanases in bleaching: from an idea to the industry,” FEMS Microbiology Reviews, vol. 13, no. 2-3, pp. 335–350, 1994.
[4]
L. Tremblay and F. Archibald, “Production of a cloned xylanase in Bacillus cereus and its performance in kraft pulp prebleaching,” Canadian Journal of Microbiology, vol. 39, no. 9, pp. 853–860, 1993.
[5]
L. Viikari, M. Ranua, A. Kantelinen, J. Sundiquist, and M. Linko, “Bleaching with enzymes,” in Biotechnology in the Pulp and Paper Industry: The Third International Conference, p. 67, Swedish Forest Products Research Laboratory, Stockholm, Sweden, 1986.
[6]
A. Gessesse and G. Mamo, “Purification and characterization of an alkaline xylanase from alkaliphilic Micrococcus sp. AR-135,” Journal of Industrial Microbiology and Biotechnology, vol. 20, no. 3-4, pp. 210–214, 1998.
[7]
M. L. Niku-Paavola, M. Ranua, A. Suurnakki, and A. Kantelinen, “Effects of lignin-modifying enzymes on pine kraft pulp,” Bioresource Technology, vol. 50, no. 1, pp. 73–77, 1994.
[8]
A. M. Madlala, S. Bissoon, S. Singh, and L. Christov, “Xylanase-induced reduction of chlorine dioxide consumption during elemental chlorine-free bleaching of different pulp types,” Biotechnology Letters, vol. 23, no. 5, pp. 345–351, 2001.
[9]
P. Bajpai, “Application of enzymes in the pulp and paper industry,” Biotechnology Progress, vol. 15, no. 2, pp. 147–157, 1999.
[10]
P. Bajpai and P. K. Bajpai, “Development of a process for the production of dissolving kraft pulp using xylanase enzyme,” Appita Journal, vol. 54, no. 4, pp. 381–384, 2001.
[11]
K. K. Y. Wong, L. U. L. Tan, and J. N. Saddler, “Multiplicity of beta-1,4-xylanase in microorganisms: functions and applications,” Microbiological Reviews, vol. 52, no. 3, pp. 305–317, 1988.
[12]
R. C. Kuhad and A. Singh, “Lignocellulose biotechnology: current and future prospects,” Critical Reviews in Biotechnology, vol. 13, no. 2, pp. 151–172, 1993.
[13]
R. C. Kuhad, A. Singh, and K. E. L. Eriksson, “Microorganisms and enzymes involved in the degradation of plant fiber cell walls,” Advances in Biochemical Engineering/Biotechnology, vol. 57, pp. 45–125, 1997.
[14]
Q. K. Beg, M. Kapoor, L. Mahajan, and G. S. Hoondal, “Microbial xylanases and their industrial applications: a review,” Applied Microbiology and Biotechnology, vol. 56, no. 3-4, pp. 326–338, 2001.
[15]
R. D. S. Khandeparkar and N. B. Bhosle, “Isolation, purification and characterization of the xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation,” Enzyme and Microbial Technology, vol. 39, no. 4, pp. 732–742, 2006.
[16]
G. L. Miller, “Use of dinitrosalicylic acid reagent for determination of reducing sugar,” Analytical Chemistry, vol. 31, no. 3, pp. 426–428, 1959.
[17]
O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall, “Protein measurement with the Folin phenol reagent,” The Journal of biological chemistry, vol. 193, no. 1, pp. 265–275, 1951.
[18]
Anon, TAPPI Test Methods, TAPPI, Atlanta, Ga, USA, 1996.
[19]
N. Gupta, V. S. Reddy, S. Maiti, and A. Ghosh, “Cloning, expression, and sequence analysis of the gene encoding the alkali-stable, thermostable endoxylanase from alkalophilic, mesophilic Bacillus sp. strain NG-27,” Applied and Environmental Microbiology, vol. 66, no. 6, pp. 2631–2635, 2000.
[20]
U. Gupta and R. Kar, “Optimization and scale up of cellulase free endo xylanase production by solid state fermentation on corn cob and by immobilized cells of a thermotolerant bacterial isolate,” Jordan Journal of Biological Sciences, vol. 1, pp. 129–134, 2008.
[21]
M. K. Kang, P. J. Maeng, and Y. H. Rhee, “Purification and characterization of two xylanases from alkalophilic Cephalosporium sp. strain RYM-202,” Applied and Environmental Microbiology, vol. 62, no. 9, pp. 3480–3482, 1996.
[22]
I. M. Mathrani and B. K. Ahring, “Thermophilic and alkalophilic xylanases from several Dictyoglomus isolates,” Applied Microbiology and Biotechnology, vol. 38, no. 1, pp. 23–27, 1992.
[23]
H. K. Sreenath and R. Joseph, “Purification and properties of extracellular xylan hydrolases of Streptomyces exfoliatus,” Folia Microbiologica, vol. 27, no. 2, pp. 107–115, 1982.
[24]
A. Gessesse, “Purification and properties of two thermostable alkaline xylanases from an alkaliphilic Bacillus sp.,” Applied and Environmental Microbiology, vol. 64, no. 9, pp. 3533–3535, 1998.
[25]
M. Ujiie, C. Roy, and M. Yaguchi, “Low-molecular-weight xylanase from Trichoderma viride,” Applied and Environmental Microbiology, vol. 57, no. 6, pp. 1860–1862, 1991.
[26]
T. Kanda, Y. Amano, and K. Nisizawa, “Purification and properties of two endo-1,4-β-xylanases from Irpex lacteus (Polyporus tulipiferae),” Journal of Biochemistry, vol. 98, no. 6, pp. 1545–1554, 1985.
[27]
T. Anthony, K. C. Raj, A. Rajendran, and P. Gunasekaran, “High molecular weight cellulase-free xylanase from alkali-tolerant Aspergillus fumigatus AR1,” Enzyme and Microbial Technology, vol. 32, no. 6, pp. 647–654, 2003.
[28]
S. P. George, A. Ahmad, and M. B. Rao, “A novel thermostable xylanase from Thermomonospora sp.: influence of additives on thermostability,” Bioresource Technology, vol. 78, no. 3, pp. 221–224, 2001.
[29]
X. T. Li, Z. Q. Jiang, L. T. Li et al., “Characterization of a cellulase-free, neutral xylanase from Thermomyces lanuginosus CBS 288.54 and its biobleaching effect on wheat straw pulp,” Bioresource Technology, vol. 96, no. 12, pp. 1370–1379, 2005.
[30]
G. Elegir, M. Sykes, and T. W. Jeffries, “Differential and synergistic action of Streptomyces endoxylanases in prebleaching of kraft pulps,” Enzyme and Microbial Technology, vol. 17, no. 10, pp. 954–959, 1995.
[31]
A. Suurnakki, A. Kantelinen, J. Buchet, and L. Vikari, “Enzyme aided bleaching of industrial softwood kraft pulps,” TAPPI Journal, vol. 77, pp. 111–116, 1994.
[32]
J. Angayarkanni, M. Palaniswamy, B. V. Pradeep, and K. Swaminathan, “Biochemical substitution of fungal xylanases for prebleaching of hardwood kraft pulp,” African Journal of Biotechnology, vol. 5, no. 10, pp. 921–929, 2006.