Publication:
Identification and characterization of ferulic acid esterase from Penicillium chrysogenum 31B: de-esterification of ferulic acid decorated with L-arabinofuranoses and D-galactopyranoses in sugar beet pectin

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Issued Date

2019

Resource Type

Article

File Type

application/pdf

ISSN

1410229

DOI

10.1016/j.enzmictec.2019.109380

Other identifier(s)

2-s2.0-85069547128

Rights Holder(s)

Scopus

Bibliographic Citation

Enzyme and Microbial Technology. Vol 131, (2019)

Suggested Citation

Phuengmaung P., Sunagawa Y., Makino Y., Kusumoto T., Handa S., Sukhumsirichart W., Sakamoto T. Identification and characterization of ferulic acid esterase from Penicillium chrysogenum 31B: de-esterification of ferulic acid decorated with L-arabinofuranoses and D-galactopyranoses in sugar beet pectin. Enzyme and Microbial Technology. Vol 131, (2019). doi:10.1016/j.enzmictec.2019.109380 Retrieved from: https://hdl.handle.net/20.500.14740/5069

Title

Identification and characterization of ferulic acid esterase from Penicillium chrysogenum 31B: de-esterification of ferulic acid decorated with L-arabinofuranoses and D-galactopyranoses in sugar beet pectin

Author(s)

Phuengmaung P.
 Sunagawa Y.
 Makino Y.
 Kusumoto T.
 Handa S.
 Sukhumsirichart W.
 Sakamoto T.

Abstract

We previously described the fungus Penicillium chrysogenum 31B, which has high performance to produce the ferulic acid esterase (FAE) for de-esterifying ferulic acids (FAs) from sugar beet pulp. However, the characteristics of this fungus have not yet been determined. Therefore, in this study, we evaluated the molecular characteristics and natural substrate specificity of the Pcfae1 gene from Penicillium chrysogenum and examined its synergistic effects on sugar beet pectin. The Pcfae1 gene was cloned and overexpressed in Pichia pastoris KM71H, and the recombinant enzyme, named PcFAE1, was characterized. The 505 amino acids of PcFAE1 possessed a GCSTG motif (Gly164 to Gly168), characteristic of the serine esterase family. By comparing the amino acid sequence of PcFAE1 with that of the FAE (AoFaeB) of Aspergillus oryzae, Ser166, Asp379, and His419 were identified as the catalytic triad. PcFAE1 was purified through two steps using anion-exchange column chromatography. Its molecular mass without the signal peptide was 75 kDa. Maximum PcFAE1 activity was achieved at pH 6.0–7.0 and 50 °C. The enzyme was stable up to 37 °C and at a pH range of 3–8. PcFAE1 activity was only inhibited by Hg2+, and the enzyme had activity toward methyl FA, methyl caffeic acid, and methyl p-coumaric acid, with specific activities of 6.97, 4.65, and 9.32 U/mg, respectively, but not on methyl sinapinic acid. These results indicated that PcFAE1 acted similar to FaeB type according the Crepin classification. PcFAE1 de-esterified O-[6-O-feruloyl-β-D-galactopyranosyl-(1→4)]-D-galactopyranose, O-[2-O-feruloyl-α-L-arabinofuranosyl-(1→5)]-L-arabinofuranose, and O-[5-O-feruloyl-α-L-arabinofuranosyl-(1→3)]-O-β-D-xylopyranosyl-(1→4)-D-xylopyranose, indicating that the enzyme could de-esterify FAs decorated with both β-D-galactopyranosidic and α-L-arabinofuranosidic residues in pectin and xylan. PcFAE1 acted in synergy with endo-α-1,5-arabinanase and α-L-arabinofuranosidase, which releases FA linked to arabinan, to digest the sugar beet pectin. Moreover, when PcFAE1 was allowed to act on sugar beet pectin together with Driselase, approximately 90% of total FA in the substrate was released. Therefore, PcFAE1 may be an interesting candidate for hydrolysis of lignocellulosic materials and could have applications as a tool for production of FA from natural substrates. © 2019 Elsevier Inc.

Subject(s)

Amino acids
 Aspergillus
 Cloning
 Column chromatography
 Enzyme activity
 Esterification
 Genes
 Substrates
 Sugar beets
 Ferulic acid esterase
 Ferulic acids
 Penicillium chrysogenum
 Sugar beet pectins
 Synergistic action
 Esters
 Alpha arabinofuranosidase
 Amino acid
 Esterase
 Ferulic acid
 Ferulic acid esterase
 Methyl caffeic acid
 Para coumaric acid
 Pectin
 Recombinant enzyme
 Serine proteinase
 Signal peptide
 Sinapic acid
 Unclassified drug
 Xylan
 Arabinofuranose
 Arabinose
 Carboxylesterase
 Coumaric acid
 Ferulic acid
 Feruloyl esterase
 Galactose
 Pectin
 Amino acid sequence
 Anion exchange
 Article
 Aspergillus oryzae
 Catalysis
 Column chromatography
 Control
 Controlled study
 Enzyme activity
 Enzyme specificity
 Esterification
 High performance liquid chromatography
 Komagataella pastoris
 Molecular weight
 Nonhuman
 Nucleotide sequence
 Penicillium chrysogenum
 PH
 Sugar beet
 Sugar beet pulp
 Synergistic effect
 Chemistry
 Enzyme stability
 Enzymology
 Gene expression
 Genetics
 Isolation and purification
 Metabolism
 Molecular cloning
 Pichia
 Temperature
 Arabinose
 Carboxylic Ester Hydrolases
 Cloning, Molecular
 Coumaric Acids
 Enzyme Stability
 Galactose
 Gene Expression
 Hydrogen-Ion Concentration
 Pectins
 Penicillium chrysogenum
 Pichia
 Substrate Specificity
 Temperature

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https://hdl.handle.net/20.500.14740/5069

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