, Volume 250, Issue 6, pp 2063–2082 | Cite as

Characterization of feruloyl esterases in maize pollen

  • Marcia M. de O BuanafinaEmail author
  • M. Fernanda Buanafina
  • Tatiana Laremore
  • Erica A. Shearer
  • Howard W. Fescemyer
Original Article

Main conclusion

Ferulic acid esterases have been identified and partially purified from maize pollen. Results suggest that maize pollen FAEs may play an important role in pollen fertilization.


A critical step in maize (Zea mays) seed production involves fertilization of the ovule by pollen, a process that relies on ability of the pollen tube to grow through the highly structured and feruloylated arabinoxylan/cellulose-rich tissue of the silk and stigma. It is known that different cell wall hydrolases are present on the surface of pollen. An important hydrolase reported to date is an endo-xylanase (ZmXYN1). We report presence and characterization of another hydrolase, ferulic acid esterase (FAE), in maize pollen. Using a combination of biochemical approaches, these FAEs were partially purified and characterized with respect to their biochemical properties and putative sequences. Maize pollen FAEs were shown to be expressed early during pollen development, to release significant amounts of both monomeric and dimeric ferulates esterified from maize silks and other grass cell walls, and to synergize with an externally applied fungal endo-1,4-β-xylanase on the release of cell wall ferulates and diferulates. Preliminary analysis of maize silk cell walls following pollination, showed a significant reduction of esterified ferulates up to 96 h following pollination, compared to unpollinated silks. These results suggest that maize pollen FAEs may play an important biological role in pollen fertilization and possibly in seed production.


Pollen ferulic acid esterase Stigma cell wall Feruloylation Zea mays Maize fertilization Ferulic acid Biodegradability 





Ferulic acid


Hydroxycinnamic acid


High performance liquid chromatography


Size-exclusion chromatography


Ferulic acid esterase


trans p-Coumaric acid


trans-Ferulic acid


cis-Ferulic acid

5-5´ DFA

5-5´-diferulic acid

8-0-4´ DFA

8-0-4´-diferulic acid

8-5c DFA

8-5cyc-diferulic acid benzofuran


8-5′-diferulic acid (open form)


Ethyl ferulate


Methyl ferulate


Methyl caffeate


Methyl syringate


Methyl vanillate


Methyl sinapate


Methyl coumarate


Feruloylated arabinoxylan trisaccharide (2-[5-O-(trans- feruloyl)-L-arabinofuranosyl]-(1 → 3)-O-ß-d-xylopyranosyl-(l → 4)-d-xylopyranose


Feruloylated arabinoxylan tetrasaccharide O-ß-d-xylopyranosyl-(1 → 4)-0-[5-O-(trans-feruloyl-α-L-arabinofuranosyl-(l → 3)]-O-ß-xylopyranosyl-(l → 4)-d xylopyranose


Feruloylated glucose disaccharide O-(4-O-trans-feruloyl-α-d-xylopyranosyl)-(1 → 6)-d-glucopyranose


p-Coumaroyl trisaccharide O-[5-O-(trans-p-coumaroyl)-α-l-arabinofuranosyl]-(1 → 3)-O-β-d-xylopyranosyl-(1 → 4)-d-xylopyranose


Feruloylated arabinan disaccharide O-(2-O-trans- feruloyl-α-L-arabinofuranosyl)-(1 → 5)-L-arabinofuranose


Feruloylated arabinan trisaccharide O-α-L-arabinofuranosyl)-(1 → 3)-O-(2-O-trans-α-L-arabinofuranosyl)-(1 → 4)-d-xylopyranose (these oligosaccharides were prepared from cell wall of sugar beet and provided by Dr Tadashi Ishii (Ishii et al. 1990; Ishii and Hiroi 1991; Ishii and Tobita 1993; Ishii 1994))


Days after emergence


4-(2-Aminoethyl)benzenesulfonyl fluoride (AEBSF)



This research was supported by the U.S. Department of Agriculture (USDA), grant number 2009-35318-0513, Department of Energy (DOE) Plant Feedstock Genomics Research Program, grant number DE-FG02-08ER64701 to MB. Small part of funding support for this work was by the Elberly College of Science, PSU. We thank Dr. T. Ishii (Japan) for supplying the feruloylated oligosaccharides (FAXX)sb, Dr Scott Borneman, Genencor, USA, for supplying feruloylated polysaccharides (FAXX)w, FAXXX, FXG, pCAXX, FAA and FAAA; Dr. B. Minkenberg (The Pennsylvania State University) for supplying rice plants for pollen collection, Dr. P. Adler and Mr. M. Mayers (USDA-ARS) for supplying big bluestem and Indian grasses for pollen collection, Mr. A. Omeis for supplying pollen from ornamental plants, Dr. M.C. Saha (Samuel Roberts Noble Foundation) for supplying the tall fescue genotype for pollen collection, Mr. P. Mush and Dr. D. Thorogood (IBERS/UK) for supplying Lolium plants for pollen collection, and Dr. P. Morris for discussions on the manuscript.

Supplementary material

425_2019_3288_MOESM1_ESM.docx (18 kb)
Supplementary material 1 (DOCX 17 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Marcia M. de O Buanafina
    • 1
    Email author
  • M. Fernanda Buanafina
    • 1
  • Tatiana Laremore
    • 2
  • Erica A. Shearer
    • 1
  • Howard W. Fescemyer
    • 1
  1. 1.Department of Biology, 208 Mueller LaboratoryThe Pennsylvania State UniversityUniversity ParkUSA
  2. 2.Penn State Proteomics and Mass Spectrometry Core Facility, The Pennsylvania State UniversityUniversity ParkUSA

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