Cloning，Expression and Characterization of Acetyl Xylan Esterase from Streptomyces griseus

doi:10.13560/j.cnki.biotech.bull.1985.2015.02.023

Abstract

Abstract: Primers were designed by putative axe annotated from the whole genome sequence of Streptomyces griseus. The axe gene was firstly cloned and linked to the prokaryotic expression vector, expressed recombinant protein induced with IPTG and purified with the Ni-NTA affinity chromatography. The results showed the cloned gene axe contained an open reading frame of 1 008 bp encoding 336 amino acid residues. The transformant harboring pET28a-Sgraxe was expressed with a protein molecular weight of 37 kD by SDS-PAGE analysis. The results indicated that the purified recombinant enzyme exhibited optimum activity at pH8.0 and 50℃, high thermal stability and strong alkali resistance range. Metal ions on recombinant enzyme activity was inhibited especially Zn²⁺. The analysis of recombinant enzymatic properties reveals potential application in industry.

Key words: Streptomyces griseus, acetyl xylan esterase, prokaryotic expression, purification, enzymatic properties

Liu Weina, Liang Di, Wang Xiaoyu, Yu Wangning, Hou Lingyu, Jin Yi, Xie Xiangming. Cloning，Expression and Characterization of Acetyl Xylan Esterase from Streptomyces griseus[J]. Biotechnology Bulletin, 2015, 31(2): 153-159.

References

[1]Saha B. Hemicellulose bioconversion [J]. J Ind Microbiol Biot, 2003, 30（5）：279-291.
[2]Prade RA. Xylanases：from biology to biotechnology [J]. Biotechnol Genet Eng, 1995, 13（12）：101-131.
[3]Berens S, Kaspari H, Klemme JH. Purification and characterization of two different xylanases from the thermophilic actinomycete Microtetraspora flexuosa SIIX. [J]. Anton Leeuw Int J G, 1996, 69（3）：235-241.
[4]Pérez J, Mu?oz-Dorado T, de la Rubia J, et al. Biodegradation and biological treatments of cellulose, hemicellulose and lignin：an overview [J]. Int Microbiol, 2002, 5（2）：53-63.
[5]Biely P, MacKenzie CR, Puls J, et al. Cooperativity of esterases and xylanases in the enzymatic degradation of acetyl xylan [J]. Nat Biotechol, 1986, 4（8）：731-733.
[6]Bouveng HO, Garegg PJ, Lindberg B. Position of the O-acetyl group in birch xylan [J]. Acta Chemical Scandinavica, 1960, 14：742-748.
[7]Bacon JSD, Gordon AH, Morris EJ. Acetyl groups in cell-wall preparations from higher plants [J]. Biochem J, 1975, 149：485-487.
[8]McDermid KP, Fosberg CW, Mackenzie CR. Purification and properties of an acetyl xylan esterase from Fibrobacter succinogenes S85[J]. Appl Environ Microb, 1990, 56（12）：3805-3810.
[9]Poutanen K, Sundberg M, Korte H, et al. Deacetylation of xylans by acetyl esterases of Trichoderma reesei [J]. Appl Environ Microb, 1990, 33：506-510.
[10]Cybinski DH, Layton I, Lowry JB, et al. An acetylxylan esterase and a xylanase expressed from genes cloned from the ruminal fungus Neocallimastix patriciarum act synergistically to degrade acetylated xylans [J]. Appl Microbiol Biot, 1999, 52：221-225.
[11]Bachmann SL, McCarthy AJ. Purification and cooperative activity of enzymes constituting the xylan-degrading system of Thermomonospora fusca [J]. Appl Environ Microb, 1991, 57（8）：2121-2130.
[12] Chungool W, Thongkam W, Raweesri P, et al. Production, purification, and characterization of acetyl esterase from Streptomyces sp. PC22 and its action in cooperation with xylanolytic enzymes on xylan degradation [J]. World J Microb Biot, 2008, 24：549-556.
[13]Biely P, Puls J, Schneider H. Acetyl xylan esterases in fungal cellulolytic systems [J]. FEBS Letters, 1985, 186（1）：80-84.
[14]Biely P, Cote GL, Kremnicky L, et al. Action of acetylxylan esterase from Trichoderma reesei on acetylated methyl glycosides [J]. FEBS Lett, 1997, 420（2）：121-124.
[15] Khan AW, Lamb KA, Overend RP. Comparison of natural hemicellulose and chemically acetylated xylan as substrates for the determination of acetyl-xylan esterase activity in Aspergilli [J]. Enzyme Microb Tech, 1990, 12（2）：127-131.
[16]Chávez R, Bull P, Eyzaguirre J. The xylanolytic enzyme system from the genus Penicillium [J]. J Biotechnol, 2006, 123（4）：413-433.
[17]Lee H, To RJ, Latta RK, et al. Some properties of extracellular acetylxylan esterase produced by the yeast Rhodotorula mucilaginosa [J]. Appl Environ Microb, 1987, 53：2831-2834.
[18]Li XL, Skory CD, Cotta MA, et al. Novel family of carbohydrate esterases, based on identification of the Hypocrea jecorina acetyl esterase gene [J]. Appl Environ Microb, 2008, 74（24）：7482-7489.
[19]Liu X, Ding S. Molecular characterization of a new acetyl xylan esterase（AXEII） from edible straw mushroom Volvariella volvacea with both de-O-acetylation and de-N-acetylation activity [J]. FEMS Microbiol Lett, 2009, 295：50-56.
[20] Pai CK, Wu ZY, Chen MJ, et al. Molecular cloning and characterization of a bifunctional xylanolytic enzyme from Neocallimastix patriciarum [J]. Appl Microbiol Biot, 2010, 85（5）：1451-1462.
[21]Dupont C, Daigneault N, Shareck F, et al. Purification and characterization of an acetyl xylan esterase produced by Streptomyces lividans [J]. Biochem J, 1996, 319：881-886.
[22]McDermid KP, MacKenzie CR, Forsberg CW. Esterase activities of Fibrobacter succinogenes subsp. succinogenes S85 [J]. Appl Environ Microb, 1990, 56（1）：127-132.
[23]Krastanova I, Guarnaccia C, Zahariev S, et al. Heterologous expression, purification, crystallization, X-ray analysis and phasing of the acetyl xylan esterase from Bacillus pumilus [J]. BBA-Proteins Proteom, 2005, 1748（2）：222-230.
[24]谢响明, 孙晓霞, 吴玉英, 等.绿色糖单孢菌产木聚糖酶规律及其耐碱耐热性的初步研究[J].生命科学研究, 2005, 9（1）：55-59.
[25]Shao W, Wiegel J. Purification and characterization of two acetyl xylan esterases from Thermoanaerobacterium sp. strain JW/SL-YS485 [J]. Appl Environ Microb, 1995, 61：729-733.
[26] Van Lieshout J, Faijes M, Nieto J, et al. Hydrolase and glycosynthase activity of endo-1, 3-glucanase from the thermophile Pyrococcus furiosus [J]. Archaea, 2004, 1（4）：285-292.
[27]Lineweaver H, Burk D. The determination of enzyme dissociation constants [J]. J Am Chem Soc, 1934, 56（3）：658-666.
[28]Zhang J, Siikaaho M, Tenkanen M, et al. The role of acetyl xylan esterase in the solubilization of xylan and enzymatic hydrolysis of wheat straw and giant reed [J]. Biotechnol Biofuels, 2011, 4（1）：60-68.
[29]Beg QK, Kapoor M, Mahajan L, et al. Microbial xylanases and their industrial applications：a review [J]. Appl Microbiol Biot, 2001, 56：326-338.
[30] Degrassi G, Okeke BC, Bruschi CV, et al. Purification and characterization of an acetyl xylan esterase from Bacillus pumilus [J]. Appl Environ Microb, 1998, 64（2）：789-792.
[31] Halgasova N, Kutejova E, Timko J. Purification and some characteristics of the acetylxylan esterase from Schizophyllum commune [J]. Biochem J, 1994, 298：751-755.
[32]Kormelink FJM, Lefebvre B, Strozyk F, et al. Purification and characterization of an acetyl xylan esterase from Aspergillus niger [J]. J Biotechnol, 1993, 27（3）：267-282.
[33]Levisson M, Han GW, Deller MC, et al. Functional and structural characterization of a thermostable acetyl esterase from Thermotoga maritime [J]. Proteins, 2012, 80（6）：1545-1559.
[34]Yoshida S, Mackie RI, Cann IKO. Biochemical and domain analyses of FSUAxe6B, a modular Acetyl xylan esterase, identify a unique carbohydrate binding module in Fibrobacter succinogenes S85 [J]. J Bacteriol, 2010, 192（2）：483-493.