Cellulose

, Volume 23, Issue 3, pp 1565–1572 | Cite as

Assessment of endoglucanase activity by analyzing the degree of cellulose polymerization and high-throughput analysis by near-infrared spectroscopy

  • Yoshiki Horikawa
  • Tomoya Imai
  • Kentaro Abe
  • Keita Sakakibara
  • Yoshinobu Tsujii
  • Asako Mihashi
  • Yoshinori Kobayashi
  • Junji Sugiyama
Original Paper

Abstract

We developed a novel and practical assessment technique for endoglucanase (EG) activity by measuring the degree of polymerization (DP) of cellulose from Eucalyptus globulus. This evaluation method demonstrated that EG II from Trichoderma reesei had higher endoglucanase activity than EG I, which has not been detected in conventional experiments using carboxymethyl cellulose as a model substrate. In addition, a high-throughput protocol for DP measurement was developed by using near-infrared spectroscopy combined with a multivariate analysis. Interpreting the regression coefficient, a reciprocal correlation was observed between the relative crystallinity of the cellulosic residue after enzymatic hydrolysis and the DP.

Keywords

Endoglucanase Cellulose microfibril Degree of polymerization NIR spectroscopy Relative crystallinity 

Abbreviations

DP

Degree of polymerization

EG

Endoglucanase

CBH

Cellobiohydrolase

BGL

β-glucosidase

CMC

Carboxymethyl cellulose

NIR

Near-infrared spectroscopy

DNS

3,5-dinitrosalicylic acid

THF

Tetrahydrofuran

GPC

Gel permeation chromatography

TGS

Triglycine sulfate

PLS

Partial least squares

Rc2

Coefficient of determination for calibration

RMSEC

Root-mean-square error of calibration

Rp2

Coefficient of determination for prediction

RMSEP

Root-mean-square error of prediction

PMOs

Polysaccharide monooxygenases

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Yoshiki Horikawa
    • 1
    • 2
  • Tomoya Imai
    • 2
  • Kentaro Abe
    • 2
  • Keita Sakakibara
    • 3
  • Yoshinobu Tsujii
    • 3
  • Asako Mihashi
    • 4
  • Yoshinori Kobayashi
    • 4
  • Junji Sugiyama
    • 2
    • 5
  1. 1.Faculty of AgricultureTokyo University of Agriculture and TechnologyFuchuJapan
  2. 2.Research Institute for Sustainable Humanosphere (RISH)Kyoto UniversityUjiJapan
  3. 3.Institute for Chemical ResearchKyoto UniversityUjiJapan
  4. 4.Tsukuba Research LaboratoryJapan Bioindustry AssociationTsukubaJapan
  5. 5.CREST-JSTKawaguchiJapan

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