ORIGINAL ARTICLE An isolated Amycolatopsis sp. GDS for cellulase and xylanase production using agricultural waste biomass S.D. Kshirsagar 1, *, G.D. Saratale 2, *, R.G. Saratale 3 , S.P. Govindwar 4 and M.K. Oh 2 1 Department of Biotechnology, Shivaji University, Kolhapur, Maharashtra, India 2 Department of Chemical and Biological Engineering, Korea University, Seongbuk-gu, Seoul, South Korea 3 Department of Environmental Science and Engineering, Ewha Womans University, Seoul, South Korea 4 Department of Biochemistry, Shivaji University, Kolhapur, Maharashtra, India Keywords Amycolatopsis sp. GDS, cellulase and xylanase, enzymatic hydrolysis, ethanol fermentation, halotolerance, wheat straw. Correspondence Min-Kyu Oh, Department of Chemical & Bio- logical Engineering, Korea University, Seoul 136–713, South Korea. E-mail: mkoh@korea.ac.kr *Both authors contributed equally to this manuscript. 2015/1244: received 25 June 2015, revised 8 October 2015 and accepted 20 October 2015 doi:10.1111/jam.12988 Abstract Aim: The aim of this study was to evaluate an isolate of Amycolatopsis sp. GDS for cellulase and xylanase production, their characterization, and its application to the preparation of biomass feedstock for ethanol production. Methods and Results: A novel potent cellulolytic bacterial strain was isolated and identified as Amycolatopsis sp. GDS. The strain secreted high levels of cellulase and xylanase in the presence of agricultural waste biomass. The enzymes were thermostable and active up to 70°C. Interestingly, the enzymes were expressed well at higher NaCl (up to 25 mol l 1 ) and ionic liquid (10%) concentrations, so that they could be used during the pretreatment of biomass. Enzyme stability in the presence of organic solvents, surfactants and oxidizing agents was also noted. Crude enzymes from Amycolatopsis sp. GDS resulted in comparable saccharification (60%) of wheat straw to commercial enzymes (64%). Conclusions: The cellulolytic enzymes from Amycolatopsis sp. GDS were stable, expressed well under conditions with various chemicals, and yielded significant amounts of hydrolysates from the biomass. The high bioethanol production using yeast co-cultures with enzymatic hydrolysates highlights the significance of selecting the strain and substrate for biofuel production. Significance and Impact of the Study: This study demonstrates the importance of the isolate Amycolatopsis sp. GDS that secretes high levels of cellulase and hemicellulase by utilizing agricultural waste biomass and its application in the preparation of biomass feedstock and sequential ethanol fermentation. Introduction Cellulases consist of three major components— endoglu- canase, exoglucanase and b-glucosidase—which act syner- gistically to convert the cellulose complex of biomass into glucose (Sukumaran et al. 2009). In addition, xylanase plays an important role in the conversion of the hemicel- lulose component of biomass into xylose. The efficient conversion of the cellulose and hemicellulose fraction of biomass into fermentable sugars requires a large enzyme dosage per unit of raw material (Sharma et al. 2004; De Lima et al. 2005). The high market price of cellulase and hemicellulase makes the cellulose to ethanol conversion process expensive (Saratale et al. 2011). For this reason, significant efforts have been made to identify various alternative strategies such as cellulolytic enzyme produc- tion using micro-organisms, improving the efficiency of known cellulases and identifying new cellulases that are stable under extreme environments (Luisa and Morana 2013). The industrial applications require high stabilities of these enzymes at extreme pH, at high temperatures and in the presence of various inhibitors including organic solvents, ionic liquid and high concentration of NaCl (Alani et al. 2008; Datta et al. 2010; Saratale et al. Journal of Applied Microbiology 120, 112--125 © 2015 The Society for Applied Microbiology 112 Journal of Applied Microbiology ISSN 1364-5072