2012 ; 22(7):
|Author||Hongchen Zheng, Yihan Liu, Xiaoguang Liu, Jianling Wang, Ying Han, Fuping Lu|
|Affiliation||Key Laboratory of Industrial Fermentation Microbiology, Education Ministry of China, 300457 Tianjin, China,College of Biotechnology, Tianjin University of Science and Technology, 300457 Tianjin, China|
|Title||Isolation, Purification, and Characterization of a Thermostable Xylanase from a Novel Strain, Paenibacillus campinasensis G1-1|
J. Microbiol. Biotechnol.2012 ; 22(7):
|Abstract||High levels of xylanase activity (143.98 IU/ml) produced
by the newly isolated Paenibacillus campinasensis G1-1
were detected when it was cultivated in a synthetic
medium. A thermostable xylanase, designated XynG1-1,
from P. campinasensis G1-1 was purified to homogeneity
by Octyl-Sepharose hydrophobic-interaction chromatography,
Sephadex G75 gel-filter chromatography, and Q-Sepharose
ion-exchange chromatography, consecutively. By multistep
purification, the specific activity of XynG1-1 was up to
1,865.5 IU/mg with a 9.1-fold purification. The molecular
mass of purified XynG1-1 was about 41.3 kDa as estimated
by sodium dodecyl sulfate polyacrylamide gel electrophoresis
(SDS-PAGE). Sequence analysis revealed that XynG1-1
containing 377 amino acids encoded by 1,134 bp genomic
sequences of P. campinasensis G1-1 shared 96% homology
with XylX from Paenibacillus campinasensis BL11 and
77%~78% homology with xylanases from Bacillus sp. YA-
335 and Bacillus sp. 41M-1, respectively. The activity of
XynG1-1 was stimulated by Ca2+, Ba2+, DTT, and β-
mercaptoethanol, but was inhibited by Ni2+, Fe2+, Fe3+,
Zn2+, SDS, and EDTA. The purified XynG1-1 displayed
a greater affinity for birchwood xylan, with an optimal
temperature of 60oC and an optimal pH of 7.5. The fact
that XynG1-1 is cellulose-free, thermostable (stability at
high temperature of 70oC~80oC), and active over a wide
pH range (pH 5.0~9.0) suggests that the enzyme is
potentially valuable for various industrial applications,
especially for pulp bleaching pretreatment.|
|Keywords||Paenibacillus campinasensis, Thermostable Xylanase, Isolation, Purification, Characterization|
Bailey, M. J., P. Biely, and K. Poutanen. 1992. Laboratory testing of method for assay of xylanase activity. J. Biotechnol. 23: 257-270.
Beg, Q. K., M. Kapoor, L. Mahajan, and G. S. Hoondal. 2001. Microbial xylanases and their industrial applications: A review. Appl. Microbiol. Biotechnol. 56: 326-338.
Berg, B., B. V. Hofsten, and G. Pettersson. 1972. Growth and cellulose formation by Cellvibrio fulvus. J. Appl. Bacteriol. 35:201-214.
Bradford, M. M. 1976. Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein-dye binding. Anal. Biochem. 72: 248-254.
Claus, D. and R. C. W. Berkeley. 1986. Genus Bacillus Cohn 1872, 174AL, pp. 1105-1139. In P. H. A. Sneath, N. S. Mair, M. E. Sharpe, and J. G. Holt (eds.). Bergey’s Manual of Systematic Bacteriology, Vol. 2. Williams and Wilkins, Baltimore, USA.
Collins, T., C. Gerday, and G. Feller. 2005. Xylanase families and extremophilic xylanases. FEMS Microbiol. Rev. 29: 3-23.
Jia, Ouyang, Shen Wang, Yan Wang, Xin Li, Mu Chen, Qiang Yong, and Shiyuan Yu. 2011. Production of a Trichoderma reesei QM9414 xylanase in Pichia pastoris and its application in biobleaching of wheat straw pulp. World J. Microbiol. Biotechnol. 27: 751-758.
Kim, J. M., H. K. Park, D. Y. Yum, B. K. Hahm, D. H. Bai, and J. H. Yu. 1994. Nucleotide sequence of the pectate lyase gene from alkali-tolerant Bacillus sp. YA-14. Biosci. Biotechnol. Biochem. 58: 947-949.
Knob, A. and E. C. Carmona. 2009. Purification and characterization of two extracellular xylanases from Penicillium sclerotiorum: A novel acidophilic xylanase. Appl. Biochem. Biotechnol. 162:429-443.
Ko, C. H., W. L. Chen, C. H. Tsai, W. N. Jane, C. C. Liu, and J. Tu. 2007. Paenibacillus campinasensis BL11: A wood material-utilizing bacterial strain isolated from black liquor. Bioresour. Technol. 98: 2727-2733.
Ko, C.-H., C.-H. Tsaia, J. Tu, H.-Y. Lee, L.-T. Kua, P.-A. Kuod, and Y.-K. Lai. 2010. Molecular cloning and characterization of a novel thermostable xylanase from Paenibacillus campinasensis BL11. Process Biochem. 45: 1638-1644.
Laemmli, U. K. 1970. Cleavage of structural proteins during assembly of head of bacteriophage T4. Nature 227: 680-685.
Maalej, Ines, Ines Belhaj, Najla Fourati Masmoudi, and Hafedh Belghith. 2009. Highly thermostable xylanase of the thermophilic fungus Talaromyces thermophilus: Purification and characterization. Appl. Biochem. Biotechnol. 158: 200-212.
Menon, Gopalakrishnan, Kalpana Mody, Jitendra Keshri, and Bhavanath Jha. 2010. Isolation, purification, and characterization of haloalkaline xylanase from a marine Bacillus pumilus Strain, GESF-1. Biotechnol. Bioprocess Eng. 15: 998-1005.
Morag, E., E. A. Bayer, and R. Lamed. 1990. Relationship of cellulosomal and noncellulosomal xylanases of Clostridium thermocellum to cellulose-degrading enzymes. J. Bacteriol. 172:6098-6105.
Nakamura, S., R. Nakai, K. Namba, T. Kubo, K. Wakabayashi, R. Aono, and K. Horikoshi. 1995. Structure-function relationship of the xylanase from alkaliphilic Bacillus sp. strain 41M-1. Nucleic Acids Symp. 34: 99-100.
Pason, Patthra, Akihiko Kosugi, Rattiya Waeonukul, Chakrit Tachaapaikoon, Khanok Ratanakhanokchai, Takamitsu Arai, Yoshinori Murata, Jun Nakajima, and Yutaka Mori. 2010. Purification and characterization of a multienzyme complex produced by Paenibacillus curdlanolyticus B-6. Appl. Microbiol. Biotechnol. 85: 573-580.
Polizeli, M. L., A. C. Rizzatti, R. Monti, H. F. Terenzi, J. A. Jorge, and D. S. Amorim. 2005. Xylanases from fungi: Properties and industrial applications. Appl. Microbiol. Biotechnol. 67:577-591.
Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 3nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.
Shin, K., M. Jeya, J. K. Lee, and Y. S. Kim. 2010. Purification and characterization of a thermostable xylanase from Fomitopsis pinicola. J. Microbiol. Biotechnol. 20: 1415-1423.
Soren, Dhananjay, Mohanlal Jana, Subhabrata Sengupta, and Anil K. Ghosh. 2009. Purification and characterization of a low molecular weight endo-xylanase from mushroom Termitomyces clypeatus. Appl. Biochem. Biotechnol. 162: 373-389.
Subramaniyan, S. and P. Prema. 2000. Cellulase-free xylanases from Bacillus and other microorganisms. FEMS Microbiol. Lett. 183: 1-7.
Suzuki, M. T. and S. J. Giovannoni. 1996. Bias caused by template annealing in the amplification of mixture of 16S rRNA genes by PCR. Appl. Environ. Microbiol. 62: 625-630.
Tamura, K., J. Dudley, M. Nei, and S. Kumar. 2007. MEGA4:Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596-1599.
Techapun, C., N. Poosaran, M. Watanabe, and K. Sasaki. 2003. Thermostable and alkaline-tolerant microbial cellulase-free xylanases produced from agricultural wastes and the properties required for use in pulp bleaching bioprocesses: A review. Process Biochem. 38: 1327-1340.
Wood, P. J., J. D. Erfle, and R. M. Teather. 1988. Use of complex formation between Congo red and polysaccharide in detection and assay of polysaccharide hydrolases. Meth. Enzymol. 160: 59-74.
Zhao, Y., K. Meng, H. Luo, P. Yang, P. Shi, H. Huang, Y. Bai, and B. Yao. 2011. Cloning, expression, and characterization of a new xylanase from alkalophilic Paenibacillus sp. 12-11. J. Microbiol. Biotechnol. 21: 861-868.