Twenty-fivealkaline xylanase producing strains were isolated from Qinghai Lake side soil samples. Among these strains, QH14 produced 648.79 U/mLxylanase, and the enzymatic specific activity was 1148.56 U/mg after purification. This alkaline xylanase producing strain belongs to genus Bacillus based on16S rDNA sequencing analysis and then was designated as Bacillus sp. QH14. The alkalinexylanaseencoding gene, XynQH14, was cloned from Bacillus sp. QH14 and expressed in Escherichiacoli BL21 (DE3). The specific activity of the recombinant xylanase XynQH14 was 700.47 Umg-1 after purification by Ni-NTA affinity chromatography. The optimal tempera-ture and pH of XynQH14 were 60℃ and pH9.2, respectively. Its activity was 50% of initial activity after incubation at 55 ℃ for 1h, 80% at pH7-11 at 37 ℃ for 24 h, and 31.02% at pH11 at 50℃ after 24 h, indicating that XynQH14 isthermosta-ble and alkali-stable. These properties ofXynQH14 suggest its favorable potential applications in pulp and paper industries.
SHAN Zhi-Qiong, ZHOU Jun-Gang, ZHOU Yu-Fei, YUAN Han-Ying, LV Hong
. Isolation and characterization of an alkaline xylanasefrom a newly isolated Bacillus sp. QH14[J]. Hereditas(Beijing), 2012
, 34(3)
: 356
-365
.
DOI: 10.3724/SP.J.1005.2012.00356
[1] Gupta S, Bhushan B, Hoondal GS. Isolation, purification and characterization of xylanase from Staphylococcus sp. SG-13 and its application in biobleaching of kraft pulp. J Appl Microbiol, 2000, 88(2): 325-334.
[2] Ziaie-Shirkolaee Y, Talebizadeh A, Soltanali S. Comparative study on application of T. lanuginosus SSBP xylanase and commercial xylanase on biobleaching of non wood pulps. Bioresource Technol, 2008, 99(16): 7433-7437.
[3] Hew LI, Ravindran V, Mollah Y, Bryden WL. Influence of exogenous xylanase supplementation on apparent metabolisable energy and amino acid digestibility in wheat for broiler chickens. Anim Feed Sci Tech, 1998, 75(2): 83-92.
[4] Almirall M, Francesch M, Perez-Vendrell AM, Brufau J, Esteve-Garcia E. The differences in intestinal viscosity produced by barley and beta-glucanase alter digesta en-zyme activities and ileal nutrient digestibilities more in broiler chicks than in cocks. J Nutr, 1995, 125(4): 947-955.
[5] Choct M, Annison G. Anti-nutritive effect of wheat pen-tosans in broiler chickens: roles of viscosity and gut mi-croflora. Br Poult Sci, 1992, 33(4): 821-834.
[6] Inborr J, Puhakka J, Bakker JGM, van der Meulen J. β-glucanase and xylanase activities in stomach and ileum of growing pigs fed wheat bran based diets with and without enzyme treatment. Arch Tierernahr, 1999, 52(3): 263-274.
[7] Romanowska I, Polak J, Bielecki S. Isolation and properties of Aspergillus niger IBT-90 xylanase for bakery. Appl Microbiol Biotechnol, 2006, 69(6): 665-671.
[8] Autio K, Härkönen H, Parkkonen T, Frigård T, Poutanen K, Siikaaho M, Åman P. Effects of purified endo-β- xy-lanase and endo-β-glucanase on the structural and baking characteristics of rye doughs. Food Sci Technol, 1996, 29(1-2): 18-27.
[9] Camacho NA, Aguilar OG. Production, purification, and characterization of a low-molecular-mass xylanase from Aspergillus sp. and its application in baking. Appl Biochem Biotech, 2003, 104(3): 159-171.
[10] Bailey MJ, Biely P, Poutanen K. Interlaboratory testing of methods for assay of xylanase activity. J Biotechnol, 1992, 23(3): 257-270.
[11] Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem, 1959, 31(3): 426-428.
[12] Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ri-bosomal DNA amplification for phylogenetic study. J Bacteriol, 1991, 173(2): 697-703.
[13] von Wintzingerode F, Göbel UB, Stackebrandt E. Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis. FEMS Microbiol Rev, 1997, 21(3): 213-229.
[14] Ksiazek TG, Erdman D, Goldsmith CS, Zaki SR, Peret T, Emery S, Tong SX, Urbani C, Comer JA, Lim W, Rollin PE, Dowell SF, Ling AE, Humphrey CD, Shieh WJ, Guarner J, Paddock CD, Rota P, Fields B, DeRisi J, Yang JY, Cox N, Hughes JM, LeDuc JW, Bellini WJ, Anderson LJ, SARS Working Group. A novel coronavirus associated with severe acute respiratory syndrome. N Engl J Med, 2003, 348(20): 1953-1966.
[15] 李业. 谷氨酰胺合成酶的表达、纯化、酶学性质及腺苷化现象的研究
[学位论文]. 北京: 清华大学, 2009.
[16] Liu D, Schmid RD, Rusnak M. Functional expression of Candida antarctica lipase B in the Escherichia coli cytoplasm-a screening system for a frequently used biocatalyst. Appl Microbiol Biotechnol, 2006, 72(5): 1024-1032.
[17] Wang J, Zhang WW, Liu JN, Cao YL, Bai XT, Gong YS, Cen PL, Yang MM. An alkali-tolerant xylanase produced by the newly isolated alkaliphilic Bacillus pumilus from paper mill effluent. Mol Biol Rep, 2010, 37(7): 3297-3302.
[18] Turunen O, Vuorio M, Fenel F, Leisola M. Engineering of multiple arginines into the Ser/Thr surface of Trichoderma reesei endo-1, 4-β-xylanase II increases the thermotolerance and shifts the pH
