Please use this identifier to cite or link to this item: https://ir.swu.ac.th/jspui/handle/123456789/12243
ชื่อเรื่อง: 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
ผู้แต่ง: Phuengmaung P.
Sunagawa Y.
Makino Y.
Kusumoto T.
Handa S.
Sukhumsirichart W.
Sakamoto T.
Keywords: 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
วันที่เผยแพร่: 2019
บทคัดย่อ: 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.
URI: https://ir.swu.ac.th/jspui/handle/123456789/12243
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85069547128&doi=10.1016%2fj.enzmictec.2019.109380&partnerID=40&md5=9b630a36fa93dbe9608da5a14ddd2308
ISSN: 1410229
Appears in Collections:Scopus 1983-2021

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