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ORIGINAL RESEARCH

Xylanase pretreatment of energy cane enables facile cellulose nanocrystal isolation Ratthapong Meesupthong . Naiyasit Yingkamhaeng . Thidarat Nimchua . Phitsanu Pinmanee . Solange I. Mussatto . Bin Li . Prakit Sukyai

Received: 19 May 2020 / Accepted: 26 October 2020 Ó Springer Nature B.V. 2020

Abstract This study investigated the effect of xylanase-assisted alkaline pretreatment (XAP) on the extraction of cellulose nanocrystal (CNCs) from energy cane (EC). Different concentrations of xylanase were tested to estimate the optimal enzyme loading required for pretreatment. XAP reduced the bleaching step from 7 to 5 h, generating a material with 93% whiteness index and a higher crystallinity index (CrI) than the alkaline-bleached fibers. A combined pretreatment using xylanase at 200 U/g removed 59 and 35% of hemicellulose and lignin, respectively, that was present in the EC which was confirmed using Fourier transform infrared R. Meesupthong  N. Yingkamhaeng  P. Pinmanee  P. Sukyai (&) Biotechnology of Biopolymers and Bioactive Compounds Special Research Unit, Department of Biotechnology, Faculty of Agro-industry, Kasetsart University, 50 Ngamwongwan Road Chatuchak, Bangkok 10900, Thailand e-mail: [email protected] T. Nimchua  P. Pinmanee Enzyme Technology Laboratory, Bioresource Technology Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani 12120, Thailand

spectroscopy and scanning electron microscopy analysis. X-ray diffraction and transmission electron microscopy revealed the XAP material required a shorter time of acid hydrolysis (30 min) to generate CNCs with similar lengths and CrI values to those obtained using only the alkaline pretreatment. Therefore, the XAP offered an efficient and promising approach for the isolation of CNCs with less use of chemicals for the bleaching process and a shorter hydrolysis time. Keywords Energy cane  Xylanase  Alkaline  Pretreatment  Cellulose nanocrystals B. Li CAS Key Laboratory of Biofuel, CAS Key Laboratory of Bio-Based Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China P. Sukyai Center for Advanced Studies for Agriculture and Food (CASAF), Kasetsart University Institute for Advanced Studies, Kasetsart University, 50 Ngamwongwan Road Chatuchak, Bangkok 10900, Thailand

S. I. Mussatto Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Kemitorvet, Building 220, 2800 Kongens Lyngby, Denmark

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Cellulose

Introduction Cellulose nanocrystals (CNCs) have promising potential as a biomaterial due to their appealing intrinsic properties that include nano-dimensions, high surface area, high aspect ratio, high crystallinity and mechanical strength, low density, renewability and biodegradability (Ilyas et al. 2018). To date, the main process used to obtain CNC has been the chemical method of acid hyd