Abstract
To efficiently produce xylobiose from xylan, high-level expression of an α-l-arabinofuranosidase gene from Thermotoga maritima was carried out in Escherichia coli. A 1.5-kb DNA fragment, coding for an α-l-arabinofuranosidase of T. maritima, was inserted into plasmid pET-20b without the pelB signal sequence leader, and produced pET-20b-araA1 with 8 nt spacing between ATG and Shine–Dalgarno sequence. A maximum activity of 12 U mg−1 was obtained from cellular extract of E. coli BL21-CodonPlus (DE3)-RIL harboring pET-20b-araA1. The over-expressed α-l-arabinofuranosidase was purified 13-fold with a 94% yield from the cellular extract of E. coli by a simple heat treatment. Production of xylooligosaccharides from corncob xylan by endoxylanase and α-l-arabinofuranosidase was examined by TLC and HPLC: xylobiose was the major product from xylan at 90 °C and its proportion in the xylan hydrolyzates increased with the reaction time. Hydrolysis with in the xylanase absence of α-l-arabinofuranosidase gave only half this yield.
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References
KB Bastawde (1992) ArticleTitleXylan structure, microbial xylanases, and their mode of action World J. Microbiol. Biotechnol. 8 353–368 Occurrence Handle1:CAS:528:DyaK3sXnsFOgsw%3D%3D
JH Choi SY Lee (2004) ArticleTitleSecretory and extracellular production of recombinant proteins using Escherichia coli Appl. Microbiol. Biotechnol. 64 625–635 Occurrence Handle10.1007/s00253-004-1559-9 Occurrence Handle1:CAS:528:DC%2BD2cXjs1eiu7Y%3D Occurrence Handle14966662
BA Degnan GT Macfarlane (1991) ArticleTitleComparison of carbohydrate substrate preferences in eight species of Bifdobacteria FEMS Microbiol. Lett. 68 151–156 Occurrence Handle1:STN:280:By2C3sjmslc%3D Occurrence Handle1778437
R Huber TA Langworthy K Helmut M Thomm CR Woese UB Sleytr KO Stetter (1986) ArticleTitleThermotoga maritima sp. nov. represents a new genus of unique extremely thermophilic eubacteria growing up 90 °C Arch. Microbiol. 144 324–333 Occurrence Handle10.1007/BF00409880 Occurrence Handle1:CAS:528:DyaL28Xktlyqsr8%3D
R Nivinskas N Malys V Klausa R Vaiškunaite E Gineikiene (1999) ArticleTitlePost-transcriptional control of bacteriophage T4 gene 25 expression: mRNA secondary structure that enhances translational initiation J. Mol. Biol. 288 291–304 Occurrence Handle10.1006/jmbi.1999.2695 Occurrence Handle1:CAS:528:DyaK1MXislWrsLk%3D Occurrence Handle10329143
M Okazaki S Fujikawa N Matsumoto (1990) ArticleTitleEffect of xylooligosaccharides on growth of Bifdobacterium J. Jpn. Soc. Nutr. Food Sci. 43 395–401 Occurrence Handle1:CAS:528:DyaK3MXit1Gnsro%3D
BW Petschow RD Talbott (1990) ArticleTitleGrowth promotion of Bifdobacterium species by whey and casein fractions from human and bovine milk J. Clin. Microbiol. 28 287–292 Occurrence Handle1:CAS:528:DyaK3cXhsFOjsL4%3D Occurrence Handle2312674
RA Reeves MD Gibbs DD Morris KR Griffiths DJ Saul PL Bergquist (2000) ArticleTitleSequencing and expression of additional xylanase genes from the hyperthermophile Thermotoga maritima FjSS3B.1 Appl. Environ. Microbiol. 66 1532–1537 Occurrence Handle10.1128/AEM.66.4.1532-1537.2000 Occurrence Handle1:CAS:528:DC%2BD3cXisVWmtrk%3D Occurrence Handle10742238
P Ruile C Winterhalter W Liebl (1997) ArticleTitleIsolation and analysis of a gene encoding α-glucuronidase, an enzyme with a novel primary structure involved in the breakdown of xylan Mol. Microbiol 23 267–279 Occurrence Handle10.1046/j.1365-2958.1997.2011568.x Occurrence Handle1:CAS:528:DyaK2sXpsVOrtQ%3D%3D Occurrence Handle9044261
W Shao J Wiegel (1992) ArticleTitlePurification and charcterization of a thermostable β-xylosidase from Thermoanaerobacter ethanolicus J. Bacteriol. 174 5848–5853 Occurrence Handle1:CAS:528:DyaK3sXjsVyluw%3D%3D Occurrence Handle1522062
RA Spanjaard K Chen JR Walker J Duin (1990) ArticleTitleFrame shift suppression at tandem AGA and AGG dodons by cloned tRNA genes: assigning a codon to argU tRNA and T4 tRNAArg Nucleic Acids Res. 18 5031–5036 Occurrence Handle1:CAS:528:DyaK3cXmtVGlu7g%3D Occurrence Handle2205835
A Sunna G Antranikian (1996) ArticleTitleGrowth and production of xylanolytic enzymes by the extreme thermophilic anaerobic bacterium Thermotoga thermarum Appl. Microbiol. Biotechnol 45 671–676 Occurrence Handle10.1007/s002530050746 Occurrence Handle1:CAS:528:DyaK28Xkt1ygtr4%3D
FA Watson G Sathe JF Kane (1995) ArticleTitleEffects of a minor isoleucyl tRNA on heterologous protein translation in Escherichia coli J. Bacterial. 177 7086–7091
YM Xue ZG Mao WL Shao (2004) ArticleTitleExpression of arabinofuranosidase gene of Thermotoga maritima in Escherichia coli purification and characterization of the recombinant Ara Acta Microbiol. Sin. 44 220–225 Occurrence Handle1:CAS:528:DC%2BD2MXitVyktr4%3D
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Revisions requested 27 October 2005; Revisions received 5 September 2005
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Xue, Y., Wu, A., Zeng, H. et al. High-level Expression of an α-l-arabinofuranosidase from Thermotoga maritima in Escherichia coli for the Production of Xylobiose from Xylan. Biotechnol Lett 28, 351–356 (2006). https://doi.org/10.1007/s10529-005-5934-0
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DOI: https://doi.org/10.1007/s10529-005-5934-0