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

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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.

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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

R 2c :

Coefficient of determination for calibration

RMSEC:

Root-mean-square error of calibration

R 2p :

Coefficient of determination for prediction

RMSEP:

Root-mean-square error of prediction

PMOs:

Polysaccharide monooxygenases

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Acknowledgments

We would like to thank Ms. K. Kanai for her technical support regarding the research. This study was supported by the New Energy and Industrial Technology Development Organization (NEDO) and The Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Number 25660138 and 15K18723.

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Correspondence to Yoshiki Horikawa.

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Horikawa, Y., Imai, T., Abe, K. et al. Assessment of endoglucanase activity by analyzing the degree of cellulose polymerization and high-throughput analysis by near-infrared spectroscopy. Cellulose 23, 1565–1572 (2016). https://doi.org/10.1007/s10570-016-0927-9

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  • DOI: https://doi.org/10.1007/s10570-016-0927-9

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