Novel Trifunctional Xylanolytic Enzyme Axy43A from Paenibacillus curdlanolyticus Strain B-6 Exhibiting Endo-Xylanase, β-d-Xylosidase, and Arabinoxylan Arabinofuranohydrolase Activities.
Theaxy43Agene encoding the intracellular trifunctional xylanolytic enzyme fromPaenibacillus curdlanolyticusB-6 was cloned and expressed inEscherichia coliRecombinant PcAxy43A consisting of a glycoside hydrolase family 43 and a family 6 carbohydrate-binding module exhibited endo-xylanase, β-xylosidase, and arabinoxylan arabinofuranohydrolase activities. PcAxy43A hydrolyzed xylohexaose and birch wood xylan to release a series of xylooligosaccharides, indicating that PcAxy43A contained endo-xylanase activity. PcAxy43A exhibited β-xylosidase activity toward a chromogenic substrate,p-nitrophenyl-β-d-xylopyranoside, and xylobiose, while it preferred to hydrolyze long-chain xylooligosaccharides rather than xylobiose. In addition, surprisingly, PcAxy43A showed arabinoxylan arabinofuranohydrolase activity; that is, it released arabinose from both singly and doubly arabinosylated xylose, α-l-Araf-(1→2)-d-Xylpor α-l-Araf-(1→3)-d-Xylpand α-l-Araf-(1→2)-[α-l-Araf-(1→3)]-β-d-XylpMoreover, the combination of PcAxy43A andP. curdlanolyticusB-6 endo-xylanase Xyn10C greatly improved the efficiency of xylose and arabinose production from the highly substituted rye arabinoxylan, suggesting that these two enzymes function synergistically to depolymerize arabinoxylan. Therefore, PcAxy43A has the potential for the saccharification of arabinoxylan into simple sugars for many applications.IMPORTANCEIn this study, the glycoside hydrolase 43 (GH43) intracellular multifunctional endo-xylanase, β-xylosidase, and arabinoxylan arabinofuranohydrolase (AXH) fromP. curdlanolyticusB-6 were characterized. Interestingly, PcAxy43A AXH showed a new property that acted on both the C(O)-2 and C(O)-3 positions of xylose residues doubly substituted with arabinosyl, which usually obstruct the action of xylanolytic enzymes. Furthermore, the studies here show interesting properties for the processing of xylans from cereal grains, particularly rye arabinoxylan, and show a novel relationship between PcAxy43A and endo-xylanase Xyn10C from strain B-6, providing novel metabolic potential for processing arabinoxylans into xylose and arabinose.
School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Thitiporn
T
Teeravivattanakit
School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Sirilak
S
Baramee
School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Paripok
P
Phitsuwan
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Rattiya
R
Waeonukul
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Patthra
P
Pason
Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
Chakrit
C
Tachaapaikoon
Graduate School of Bioresources, Mie University, Tsu, Japan.
Kazuo
K
Sakka
School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand khanok.rat@kmutt.ac.th.
Khanok
K
Ratanakhanokchai
Enzyme Microb Technol. 2015 Apr;71:28-35
25765307
Appl Environ Microbiol. 2016 Jan 04;82(6):1686-92
26729713
Nature. 1970 Aug 15;227(5259):680-5
5432063
Appl Biochem Biotechnol. 2015 Feb;175(4):1960-70
25432346
J Biosci Bioeng. 2013 Aug;116(2):152-9
23474097
J Mol Biol. 2006 May 26;359(1):97-109
16631196
PLoS One. 2012;7(6):e38134
22761666
Gene. 2000 Apr 18;247(1-2):137-43
10773453
Eur J Biochem. 2000 Nov;267(22):6633-41
11054116
Biochemistry. 2005 Jan 11;44(1):387-97
15628881
Biosci Biotechnol Biochem. 2010;74(11):2358-60
21071839
Biotechnol Adv. 2000 Aug;18(5):403-23
14538102
J Microbiol Biotechnol. 2010 Dec;20(12):1711-6
21193828
Biosci Biotechnol Biochem. 1993 Feb;57(2):273-7
7763496
Nat Struct Biol. 2002 Sep;9(9):665-8
12198486
Appl Environ Microbiol. 2006 Apr;72(4):2483-90
16597947
Appl Environ Microbiol. 2009 Jul;75(13):4410-8
19395566
Biotechnol Adv. 2014 Mar-Apr;32(2):316-32
24239877
PLoS One. 2013 Sep 09;8(9):e73575
24039988
Microbiol Rev. 1988 Sep;52(3):305-17
3141761
Biochem Biophys Res Commun. 2010 Nov 26;402(4):644-50
20971079
Curr Opin Biotechnol. 2000 Apr;11(2):187-98
10753763
Appl Biochem Biotechnol. 2013 Aug;170(8):1950-64
23797510
J Biol Chem. 2014 Sep 12;289(37):25957-75
25070894
Nucleic Acids Res. 2014 Jan;42(Database issue):D490-5
24270786
Appl Biochem Biotechnol. 2014 Sep;174(1):81-92
25080375
Appl Microbiol Biotechnol. 2016 Feb;100(3):1501-10
26559526
Appl Microbiol Biotechnol. 2015 Nov;99(21):8943-54
25971196
J Gen Microbiol. 1989 Feb;135(Pt 2):285-92
2515244
J Ind Microbiol Biotechnol. 2003 May;30(5):279-91
12698321
Food Chem. 2014 Apr 1;148:381-7
24262572
Biochim Biophys Acta. 2007 Sep;1774(9):1192-8
17689155
Enzyme Microb Technol. 2015 May;72:1-9
25837501
J Biol Chem. 1951 Nov;193(1):265-75
14907713
Biotechnol Biofuels. 2013 Feb 20;6(1):27
23422003
Appl Biochem Biotechnol. 2007 Apr;141(1):51-76
17625266
J Ind Microbiol Biotechnol. 2012 Jan;39(1):143-52
21720773
Appl Microbiol Biotechnol. 2006 Dec;73(4):850-61
16944135
Appl Microbiol Biotechnol. 2005 Jun;67(5):641-7
15650848