Assessment of the enzymatic hydrolysis profile of cellulosic substrates based on reducing sugar release

M.H.L. Silveira, R.S. Aguiar, Matti Siika-aho, L.P. Ramos

Research output: Contribution to journalArticleScientificpeer-review

18 Citations (Scopus)

Abstract

The activity profile of a 1:0.30 mixture of Celluclast 1.5 L FG and Novozym 188 (Novozymes) was investigated using Whatman #1 filter paper (W1FP) as a single substrate for hydrolysis. The procedure was based on the ability of the enzymes to release total (RSTot), insoluble (RSInsol) and soluble (RSSol) reducing sugars from W1FP. RSInsol was used to estimate endoglucanase (EnG) activity whereas exoglucanases (ExG) were assessed by measuring RSSol in the presence of d-gluconolactone. Finally, the ß-glucosidase (ßG) activity was derived from the difference between RSSol measurements in the presence and absence of d-gluconolactone. When this analytical procedure was applied to W1FP using 9.64 mg mL-1 of the enzyme mixture, the relative contributions of EnG, ExG and ßG to the total cellulase activity were 63.28%, 12.02% and 24.70%, respectively. Also, this ratio changed with changes in the enzyme loading, giving a new insight into the synergy that exists among the enzymes.
Original languageEnglish
Pages (from-to)392-396
JournalBioresource Technology
Volume151
DOIs
Publication statusPublished - 2014
MoE publication typeA1 Journal article-refereed

Fingerprint

Enzymatic hydrolysis
Sugars
Cellulase
hydrolysis
sugar
Enzymes
enzyme
substrate
Substrates
filter
Glucosidases
Hydrolysis
beta-glucono-1,5-lactone

Keywords

  • cellulases
  • enzyme activity
  • filter paper
  • reducing sugars
  • synergism

Cite this

@article{d3fca4f8c51f400eb2ffa408e2a2dc38,
title = "Assessment of the enzymatic hydrolysis profile of cellulosic substrates based on reducing sugar release",
abstract = "The activity profile of a 1:0.30 mixture of Celluclast 1.5 L FG and Novozym 188 (Novozymes) was investigated using Whatman #1 filter paper (W1FP) as a single substrate for hydrolysis. The procedure was based on the ability of the enzymes to release total (RSTot), insoluble (RSInsol) and soluble (RSSol) reducing sugars from W1FP. RSInsol was used to estimate endoglucanase (EnG) activity whereas exoglucanases (ExG) were assessed by measuring RSSol in the presence of d-gluconolactone. Finally, the {\ss}-glucosidase ({\ss}G) activity was derived from the difference between RSSol measurements in the presence and absence of d-gluconolactone. When this analytical procedure was applied to W1FP using 9.64 mg mL-1 of the enzyme mixture, the relative contributions of EnG, ExG and {\ss}G to the total cellulase activity were 63.28{\%}, 12.02{\%} and 24.70{\%}, respectively. Also, this ratio changed with changes in the enzyme loading, giving a new insight into the synergy that exists among the enzymes.",
keywords = "cellulases, enzyme activity, filter paper, reducing sugars, synergism",
author = "M.H.L. Silveira and R.S. Aguiar and Matti Siika-aho and L.P. Ramos",
year = "2014",
doi = "10.1016/j.biortech.2013.09.135",
language = "English",
volume = "151",
pages = "392--396",
journal = "Bioresource Technology",
issn = "0960-8524",
publisher = "Elsevier",

}

Assessment of the enzymatic hydrolysis profile of cellulosic substrates based on reducing sugar release. / Silveira, M.H.L.; Aguiar, R.S.; Siika-aho, Matti; Ramos, L.P.

In: Bioresource Technology, Vol. 151, 2014, p. 392-396.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Assessment of the enzymatic hydrolysis profile of cellulosic substrates based on reducing sugar release

AU - Silveira, M.H.L.

AU - Aguiar, R.S.

AU - Siika-aho, Matti

AU - Ramos, L.P.

PY - 2014

Y1 - 2014

N2 - The activity profile of a 1:0.30 mixture of Celluclast 1.5 L FG and Novozym 188 (Novozymes) was investigated using Whatman #1 filter paper (W1FP) as a single substrate for hydrolysis. The procedure was based on the ability of the enzymes to release total (RSTot), insoluble (RSInsol) and soluble (RSSol) reducing sugars from W1FP. RSInsol was used to estimate endoglucanase (EnG) activity whereas exoglucanases (ExG) were assessed by measuring RSSol in the presence of d-gluconolactone. Finally, the ß-glucosidase (ßG) activity was derived from the difference between RSSol measurements in the presence and absence of d-gluconolactone. When this analytical procedure was applied to W1FP using 9.64 mg mL-1 of the enzyme mixture, the relative contributions of EnG, ExG and ßG to the total cellulase activity were 63.28%, 12.02% and 24.70%, respectively. Also, this ratio changed with changes in the enzyme loading, giving a new insight into the synergy that exists among the enzymes.

AB - The activity profile of a 1:0.30 mixture of Celluclast 1.5 L FG and Novozym 188 (Novozymes) was investigated using Whatman #1 filter paper (W1FP) as a single substrate for hydrolysis. The procedure was based on the ability of the enzymes to release total (RSTot), insoluble (RSInsol) and soluble (RSSol) reducing sugars from W1FP. RSInsol was used to estimate endoglucanase (EnG) activity whereas exoglucanases (ExG) were assessed by measuring RSSol in the presence of d-gluconolactone. Finally, the ß-glucosidase (ßG) activity was derived from the difference between RSSol measurements in the presence and absence of d-gluconolactone. When this analytical procedure was applied to W1FP using 9.64 mg mL-1 of the enzyme mixture, the relative contributions of EnG, ExG and ßG to the total cellulase activity were 63.28%, 12.02% and 24.70%, respectively. Also, this ratio changed with changes in the enzyme loading, giving a new insight into the synergy that exists among the enzymes.

KW - cellulases

KW - enzyme activity

KW - filter paper

KW - reducing sugars

KW - synergism

U2 - 10.1016/j.biortech.2013.09.135

DO - 10.1016/j.biortech.2013.09.135

M3 - Article

VL - 151

SP - 392

EP - 396

JO - Bioresource Technology

JF - Bioresource Technology

SN - 0960-8524

ER -