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Purification and characterization of an endo-xylanase from Trichoderma sp., with xylobiose as the main product from xylan hydrolysis

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Abstract

Fungal endo-β-1,4-xylanases (endo-xylanases) can hydrolyze xylan into xylooligosaccharides (XOS), and have potential biotechnological applications for the exploitation of natural renewable polysaccharides. In the current study, we aimed to screen and characterize an efficient fungal endo-xylanase from 100 natural humus-rich soil samples collected in Guizhou Province, China, using extracted sugarcane bagasse xylan (SBX) as the sole carbon source. Initially, 182 fungal isolates producing xylanases were selected, among which Trichoderma sp. strain TP3-36 was identified as showing the highest xylanase activity of 295 U/mL with xylobiose (X2) as the main product when beechwood xylan was used as substrate. Subsequently, a glycoside hydrolase family 11 endo-xylanase, TXyn11A, was purified from strain TP3-36, and its optimal pH and temperature for activity against beechwood xylan were identified to be 5.0 and 55 °C, respectively. TXyn11A was stable across a broad pH range (3.0–10.0), and exhibited strict substrate specificity, including xylan from beechwood, wheat, rye, and sugarcane bagasse, with Km and Vmax values of 5 mg/mL and 1250 μmol/mg min, respectively, toward beechwood xylan. Intriguingly, the main product obtained from hydrolysis of beechwood xylan by TXyn11A was xylobiose, whereas SBX hydrolysis resulted in both X2 and xylotriose. Overall, these characteristics of the endo-xylanase TXyn11A indicate several potential industrial applications.

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Abbreviations

CMC:

Carboxymethylcellulose

DNS:

3,5-Dinitrosalicylic acid

DTT:

Dithiothreitol

GH:

Glycoside hydrolase

HPLC:

High-performance liquid chromatography

ITS:

Internal transcribed spacer

PDA:

Potato dextrose agar

SB:

Sugarcane bagasse

SBX:

SB xylan

SMART:

Simple modular architecture research tool

Tm:

Temperature

XOS:

Xylooligosaccharides

X1:

Xylose

X2:

Xylobiose

X3:

Xylotriose

X4:

Xylotetraose

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Acknowledgements

We would like to thank Mu-Qing Zhang group from State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, China, for providing raw SB materials. This work was financially supported by Grants from the Autonomous Research Project of State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources (SKLCUSA-a201902, SKLCUSA-a201923), the Guangxi BaGui Scholars Program Foundation (Grant No. 2011A001), the Guangxi Natural Science Foundation (Grant No. 2012GXNSFGA060005), Training Program for 1000 Young and Middle-Aged Key Teachers in Guangxi at 2019, and the ‘One Hundred Person’ Project of Guangxi.

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  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi Research Center for Microbial and Enzyme Engineering Technology, College of Life Science and Technology, Guangxi University, 100 Daxue Road, Nanning, 530004, Guangxi, People’s Republic of China

    Li-Hao Fu, Nan Jiang, Cheng-Xi Li, Xue-Mei Luo, Shuai Zhao & Jia-Xun Feng

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  1. Li-Hao Fu
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  2. Nan Jiang
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Fu, LH., Jiang, N., Li, CX. et al. Purification and characterization of an endo-xylanase from Trichoderma sp., with xylobiose as the main product from xylan hydrolysis. World J Microbiol Biotechnol 35, 171 (2019). https://doi.org/10.1007/s11274-019-2747-1

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