Highly Efficient Extraction of Ferulic Acid from Cereal Brans by a New Type A Feruloyl Esterase from Eupenicillium parvum in Combination with Dilute Phosphoric Acid Pretreatment
Feruloyl esterase (FAE) is a critical enzyme in bio-extraction of ferulic acid (FA) from plant cell wall. A new FAE (EpFAE1) encoding gene was isolated from Eupenicillium parvum and heterologously expressed in Pichia pastoris cells. Based on phylogenetic tree analysis, the protein EpFAE1 belongs to type A of the seventh FAE subfamily. Using methyl ferulate as substrate, the optimum temperature and pH for the catalytic activity of EpFAE1 were 50 °C and 5.5, respectively. The enzyme exhibited high stability at 50 °C, in a wide pH range (3.0–11.0), or in the presence of 2 M of NaCl. Together with the endo-xylanase EpXYN1, EpFAE1 released 72.32% and 4.00% of the alkali-extractable FA from de-starched wheat bran (DSWB) or de-starched corn bran (DSCB), respectively. Meanwhile, the substrates were pretreated with 1.75% (for DSWB) or 1.0% (for DSCB) of phosphoric acid (PA) at 90 °C for 12 h, followed by enzymatic hydrolysis of the soluble and insoluble fractions. The release efficiencies of FA were up to 84.64% for DSWB and 66.73% for DSCB. Combined dilute PA pretreatment with enzymatic hydrolysis is a low-cost and highly efficient method for the extraction of FA from cereal brans.
KeywordsFeruloyl esterase Eupenicillium parvum Pretreatment with phosphoric acid Cereal brans Ferulic acid
feruloyl esterase (ferulic acid esterase)
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
de-starched wheat bran
de-starched corn bran
high-performance liquid chromatography
The authors thank Dr. Franz St. John from the USDA Forest Service for improving the manuscript.
This work was supported by grants from a 948 Research Project (No. 2013-4-16) from the State Forestry Administration of China, a research project (30370043) from the National Natural Science Foundation of China, the Jiangsu Provincial Government Scholarship for Overseas Studies from Jiangsu Provincial Department of Education, China, a Science and Technology Project of Guizhou Province in China (Grant No. 2333), and the Priority Academic Program Development of Jiangsu Higher Education Institutions.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
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