Applied Biochemistry and Biotechnology

, Volume 189, Issue 4, pp 1304–1317 | Cite as

Cloning, Characterization, and Functional Expression of a Thermostable Type B Feruloyl Esterase from Thermophilic Thielavia Terrestris

  • Zhen Meng
  • Qin-Zheng Yang
  • Jing-zhen Wang
  • Yun-Hua HouEmail author


Feruloyl esterases (FAEs) have great potential applications in paper and breeding industry. A new thermo-stable feruloyl esterase gene, TtfaeB was identified from the thermophilic fungus Thielavia terrestris h408. Deduced protein sequence shares the identity of 67% with FAEB from Neurospora crassa. The expression vector pPIC9K-TtfaeB was successfully constructed and electro-transformed into GS115 strain of Pichia pastoris. One transformant with high feruloyl esterase yield was obtained through plate screening and named TtFAEB1. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of fermentation supernatant from transformant TtFAEB1 showed a distinct protein band appearing at the position of about 35-kDa, indicating that TtfaeB gene has been successfully expressed in P. pastoris. The recombinant TtFAEB was purified by affinity chromatography and the specific activity of purified TtFAEB was 6.06 ± 0.72 U/mg. The optimal temperature and pH for purified recombinant TtFAEB was 60 °C and 7.0, respectively. TtFAEB was thermostable, retaining 96.89 and 84.16% of the maximum activity after being treated for 1 h at 50 °C and 60 °C, respectively. Additionally, the enzyme was stable in the pH range 4.5–8.0. The homology model of TtFAEB showed that it consists of a single domain adopting a typical α/β-hydrolase fold and contains a catalytic triad formed by Ser117, Asp201, and His260. TtFAEB in association with xylanase from Trichoderma reesei could release 77.1% of FA from destarched wheat bran. The present results indicated that the recombinant TtFAEB with excellent enzymatic properties is a promising candidate for potential applications in biomass deconstruction and biorefinery.

Key words

Feruloyl esterase Thermostable Thielavia terrestris Heterologous expression 


Funding Information

This study received financial support from National Natural Science Foundation of China (Grant No. 31570118 and 31800116), Shandong Provincial Natural Science Foundation, China (Grant No. ZR2017MC039 and ZR2015CM029).

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

12010_2019_3065_MOESM1_ESM.doc (622 kb)
ESM 1 (DOC 621 kb)


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Bioengineering, Shandong Provincial Key Laboratory of Microbial EngineeringQilu University of TechnologyJinanP. R. China
  2. 2.State Key Laboratory of Bio-based Materials and Green PapermakingJinanP. R. China

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