Extraction and Assay of Ferulic Acid Esterase From Malted Barley*

Humberstone, F.J.; Briggs, D. E.

doi:10.1002/j.2050-0416.2000.tb00036.x

Cited by 47 publications

(34 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…During germination the arabinoxylan in the cell walls of the starch-containing endosperm is broken down to allow access of hydrolytic enzymes to the starch granules and protein stored within the cells. Acetyl-and feruloyl-substituents are removed by esterases 2,14,15,19 . The core arabinoxylan polysaccharide is broken down by a number of glycoside hydrolases which include ␣-L-arabinofuranosidase (arabinosidase), endoxylanase and ␤-D-xylopyranosidase (␤-xylosidase) 20 .…”

Section: -2863(9'8-32mentioning

confidence: 99%

Purification of an Arabinofuranosidase and Two β-Xylopyranosidases from Germinated Wheat

Grant

Briggs

Fitchett

et al. 2003

Journal of the Institute of Brewing

View full text Add to dashboard Cite

Arabinosidase and ␤-xylosidase activities were detected in germinated wheat grain, and both increased over seven days of germination, under malting conditions. Arabinosidase was partially purified by anion exchange chromatography, chromatofocusing and gel filtration chromatography. The pH optimum of the partially purified enzyme was 4.2 and the K M was 1.90 mM p-NPAra. Edman degradation, MALDI-TOF mass spectrometry and nano-ESI mass spectrometry were used to identify the two major proteins in the partially purified arabinosidase mixture. The two proteins were a ␤-amylase with an amino acid sequence partially homologous to a barley ␤-amylase, and three wheat serine protease inhibitors. Further purification, by affinity chromatography and hydrophobic interaction chromatography, removed the identified contaminating proteins. At this point an 80 kDa protein was detected by SDS-PAGE. No identity could be assigned to this protein by MALDI-TOF mass spectrometry by reference to electronic protein databases. Similarly, the ␤-xylosidase was partially purified by anion exchange chromatography followed by chromatofocusing and gel filtration chromatography. The first step separated the mixture into two distinct fractions with K M values of 3.35 and 4.01 mM p-NP-Xyl and pH optima of 4.5. The latter fraction also displayed xylanase activity against RBBxylan.

show abstract

Section: -2863(9'8-32mentioning

confidence: 99%

Purification of an Arabinofuranosidase and Two β-Xylopyranosidases from Germinated Wheat

Grant

Briggs

Fitchett

et al. 2003

Journal of the Institute of Brewing

View full text Add to dashboard Cite

show abstract

“…When isolated barley aleurone layers were incubated with gibberelic acid some cell wall degradation occurred, but the ferulic acid was still attached to arabinoxylan oligomers 15,16 , and so in this case ferulic acid esterase was not active. Both soluble and insoluble esterases have been detected in barley 18 . The work reported here has been carried out on the soluble material.…”

Section: -2863(9'8-32mentioning

confidence: 99%

“…Extraction was achieved using the method previously described by Humberstone and Briggs 18 . Barley malt (3g) was ground and then finely dispersed with a Polytron homogeniser with extraction buffer (20mL; tris(hydroxymethyl)aminomethane hydrochloride [Tris\HCl], 50mM, pH 8.0; reduced glutathione, 25mM; Triton-X-100, 1% (w/v); polyvinylpolypyrrolidone [PVPP], 0.2g).…”

Section: )\Xvegxmsr Sj Jivypmg Egmh Iwxivewi Jvsq Fevpi] Qepxmentioning

confidence: 99%

See 1 more Smart Citation

Partial Purification of Ferulic Acid Esterase from Malted Barley

Humberstone¹,

Briggs²

2002

Journal of the Institute of Brewing

View full text Add to dashboard Cite

A partial purification of ferulic acid esterase, which degrades feruloyl glycerol, has been achieved from barley malt in small yields. Coloured and viscous materials were removed from the malt extract using batch-elution anion exchange chromatography. Further steps included gradient elution anion exchange chromatography and gel filtration chromatography. Estimations of the molecular weight varied greatly from 22KDa to 158 KDa, possibly because the protein interacted with the matrix of the gel exclusion chromatography column and because multiple forms of the enzyme were present. The partially purified feruloyl esterase had an apparent Km of 0.46% feruloyl glycerol. However more than one enzyme may be present and the substrate contains two isomers and so Michelis-Menten kinetics may not be appropriate.

show abstract

“…Fenolové kyseliny a anthokyany způsobují antioxidační aktivitu červených a modrých hlíz brambor, ferulová kyselina se u nich vyskytuje v množství 28 mg.kg -1 (Lachman et al, 2005). Ferulová kyselina zvyšuje hydrofobicitu arabinoxylanové molekuly a snižuje její rozpustnost (Humberstone and Briggs, 2000). Ferulová kyselina pomáhá tvořit trojrozměrnou síťovitou strukturu buněčných stěn tak, že se esterovou vazbou spojuje do dimerů (Saulnier et al, 1999) a vytváří příčné vazby polymerů buněčných stěn.…”

unclassified