title: A thermo-halo-tolerant and proteinase-resistant endoxylanase from Bacillus sp. HJ14.
abstract: A glycosyl hydrolase family 10 endoxylanase from Bacillus sp. HJ14 was grouped in a separated cluster with another six Bacillus endoxylanases which have not been characterized. These Bacillus endoxylanases showed less than 52% amino acid sequence identity with other endoxylanases and far distance with endoxylanases from most microorganisms. Signal peptide was not detected in the endoxylanase. The endoxylanase was expressed in Escherichia coli BL21 (DE3), and the purified recombinant enzyme (rXynAHJ14) was characterized. rXynAHJ14 was apparent optimal at 62.5 °C and pH 6.5 and retained more than 55% of the maximum activity when assayed at 40-75 °C, 23% at 20 °C, 16% at 85 °C, and even 8% at 0 °C. Half-lives of the enzyme were more than 60 min, approximately 25 and 4 min at 70, 75, and 80 °C, respectively. The enzyme exhibited more than 62% xylanase activity and stability at the concentration of 3-30% (w/v) NaCl. No xylanase activity was lost after incubation of the purified rXynAHJ14 with trypsin and proteinase K at 37 °C for 60 min. Different components of oligosaccharides were detected in the time-course hydrolysis of beechwood xylan by the enzyme. During the simulated intestinal digestion phase in vitro, 11.5-19.0, 15.3-19.0, 21.9-27.7, and 28.2-31.2 μmol/mL reducing sugar were released by the purified rXynAHJ14 from soybean meal, wheat bran, beechwood xylan, and rapeseed meal, respectively. The endoxylanase might be an alternative for potential applications in the processing of sea food and saline food and in aquaculture as agastric fish feed additive.
authors.affiliation: Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Yunnan Normal University, Kunming, 650500, People's Republic of China, junpeizhou@126.com.
authors.forename: Junpei
authors.initials: J
authors.lastname: Zhou
authors.forename: Qian
authors.initials: Q
authors.lastname: Wu
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Minghe
authors.initials: M
authors.lastname: Mo
authors.forename: Xianghua
authors.initials: X
authors.lastname: Tang
authors.forename: Junjun
authors.initials: J
authors.lastname: Li
authors.forename: Bo
authors.initials: B
authors.lastname: Xu
authors.forename: Junmei
authors.initials: J
authors.lastname: Ding
authors.forename: Qian
authors.initials: Q
authors.lastname: Lu
authors.forename: Zunxi
authors.initials: Z
authors.lastname: Huang
mesh_terms: D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019286:Endopeptidase K / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D059748:Proteolysis; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000378:metabolism; D013696:Temperature; D014357:Trypsin / Q000378:metabolism; D014990:Xylans / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014990:Xylans; D012965:Sodium Chloride; D043364:Endo-1,4-beta Xylanases; D014357:Trypsin; D019286:Endopeptidase K
references.citation: Extremophiles. 2001 Apr;5(2):73-83
references.pmid: 11354458
references.citation: Biotechnol Biofuels. 2011 May 18;4:12
references.pmid: 21592333
references.citation: Appl Microbiol Biotechnol. 2009 Oct;84(6):1107-15
references.pmid: 19506861
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Bioresour Technol. 2013 Jan;128:145-55
references.pmid: 23196234
references.citation: Appl Biochem Biotechnol. 2007 Apr;141(1):1-14
references.pmid: 17625262
references.citation: Br Poult Sci. 1992 Sep;33(4):821-34
references.pmid: 1393677
references.citation: FEMS Microbiol Rev. 2005 Jan;29(1):3-23
references.pmid: 15652973
references.citation: Bioresour Technol. 2013 Feb;130:161-7
references.pmid: 23306124
references.citation: J Biol Chem. 2010 Jan 22;285(4):2721-33
references.pmid: 19940147
references.citation: N Biotechnol. 2011 Oct;28(6):814-21
references.pmid: 21890005
references.citation: J Microbiol Biotechnol. 2008 Mar;18(3):410-6
references.pmid: 18388456
references.citation: Extremophiles. 2009 Sep;13(5):849-57
references.pmid: 19655217
references.citation: J Food Sci. 2012 May;77(5):C506-11
references.pmid: 22510018
references.citation: Plant J. 2001 Apr;26(2):191-204
references.pmid: 11389760
references.citation: Enzyme Microb Technol. 2010 May 5;46(6):506-12
references.pmid: 25919627
references.citation: J Ind Microbiol Biotechnol. 2012 Jul;39(7):965-75
references.pmid: 22430498
references.citation: J Anim Physiol Anim Nutr (Berl). 2012 Jun;96(3):335-64
references.pmid: 21692871
references.citation: Genome Res. 2008 May;18(5):821-9
references.pmid: 18349386
references.citation: J Microbiol Biotechnol. 2012 Apr;22(4):501-9
references.pmid: 22534297
references.citation: Genome Res. 2004 Jun;14(6):1188-90
references.pmid: 15173120
references.citation: Nucleic Acids Res. 2008 Jan;36(Database issue):D281-8
references.pmid: 18039703
references.citation: Enzyme Microb Technol. 2012 Sep 10;51(4):193-9
references.pmid: 22883553
references.citation: Biotechnol Lett. 2012 Nov;34(11):2093-9
references.pmid: 22864505
references.citation: Biotechnol Biofuels. 2013 Feb 18;6(1):26
references.pmid: 23418789
references.citation: Mol Biol Evol. 2007 Aug;24(8):1596-9
references.pmid: 17488738
references.citation: J Mol Biol. 1990 Oct 5;215(3):403-10
references.pmid: 2231712
references.citation: Appl Microbiol Biotechnol. 2008 Aug;80(2):231-40
references.pmid: 18521591