This study evaluates certain pre-treatment methods for Lantana camara stem for efficient conversion to fermentable sugars. The composition analysis of L. camara stem showed 66.8% (w/w) holocellulose, 34.9% (w/w) cellulose and 17% (w/w) hemicellulose. Comparative analysis of various chemical, physical or physico-chemical pre-treatments on L. camara stem was performed. Of all pretreatment methods used, pre-treatment with 1% (v/v) H2SO4 assisted autoclaving gave maximum total reducing sugar yield 132.7 mg/g (13.2 g/L) of raw biomass in pretreated hydrolysate. Major contribution to total reducing sugar was from hemicellulosic fraction, because total pentose sugar yield was 119.4 mg/g of raw biomass whereas, glucose released was only 10 mg/g of untreated biomass. The enzymatic saccharification of pre-treated L. camara stem by 1% (v/v) H2SO4 assisted autoclaving was performed with partially purified carboxymethylcellulase from Bacillus amyloliquefaciens SS35. Enzymatic saccharification at 30 °C for 48 h gave total reducing sugar yield, 63.3 mg/g of pre-treated biomass in the hydrolysate, while untreated biomass gave 43.3 mg/g of untreated biomass. The total sugar yield i.e. the sum of pre-treated biomass hydrolysate total reducing sugar (132.7 mg/g of raw biomass) and enzymatic hydrolysate total reducing sugar (63.3 mg/g of pre-treated biomass) was 196.0 mg/g of raw biomass, indicating the effectiveness of pre-treatment method. Field emission scanning electron microscopy, Fourier transform infrared and X-ray diffraction analyses displayed enhanced porosity, removal of non-cellulosic sugars and increased cellulose crystallinity, respectively, in pre-treated L. camara stem, showing the effectiveness of acid assisted autoclaving pre-treatment.
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Lantana camara Stem
Total reducing sugars
Carboxy methyl cellulose
Field emission scanning electron microscopy
Fourier transform infrared
Total sugar yield (sum of pre-treated hydrolysate TRS and enzymatic hydrolysate TRS)
Aditiya HB, Mahlia TMI, Chong WT, Nur H, Sebayang AH (2016) Second generation bioethanol production: a critical review. Renew Sustain Energy Rev 66:631–653
Balat M (2011) Production of bioethanol from lignocellulosic materials via the biochemical pathway: a review. Energy Convers Manag 52–2:858–875
Balat M, Balat H, Öz C (2008) Progress in bioethanol processing. Prog Energy Combust Sci 34–5:551–573
Borah AJ, Singh S, Goyal A, Moholkar VS (2016) An assessment of the potential of invasive weeds as multiple feedstocks for biofuel production. RSC Adv 6–52:47151–47163
Cao Y, Tan H (2005) Study on crystal structures of enzyme-hydrolyzed cellulosic materials by X-ray diffraction. Enzyme Microb Technol 36–2:314–317
Chandel AK, Kapoor RK, Singh AK, Kuhad RC (2007) Detoxification of sugarcane bagasse hydrolysate improves ethanol production by Candida shehatae NCIM 3501. Bioresour Technol 98:1947–1950
Das SP, Ghosh A, Gupta A, Goyal A, Das D (2013) Lignocellulosic fermentation of wild grass employing recombinant hydrolytic enzymes and fermentative microbes with effective bioethanol recovery. Biomed Res Int 2013–386063:14
Day MD, Wiley CJ, Playford J, Zalucki MP (2003) Lantana: current management status and future prospects. ACIAR Canberra 435–2016:33733
Gupta R, Mehta G, Khasa YP, Kuhad RC (2011) Fungal delignification of lignocellulosic biomass improves the saccharification of cellulosics. Biodegradation 22–4:797–804
Hahn-Hägerdal B, Galbe M, Gorwa-Grauslund MF, Lidén G, Zacchi G (2006) Bio-ethanol-the fuel of tomorrow from the residues of today. Trends Biotechnol 24–12:549–556
Hall M, Bansal P, Lee JH, Realff MJ, Bommarius AS (2010) Cellulose crystallinity-a key predictor of the enzymatic hydrolysis rate. FEBS J 277–6:1571–1582
Hammond JB, Kruger NJ (1988) The bradford method for protein quantitation New protein techniques. Humana Press, Totowa, pp 25–32
Hendriks ATWM, Zeeman G (2009) Pre-treatments to enhance the digestibility of lignocellulosic biomass. Bioresour Technol 1001:10–18
Ibrahim MM, El-Zawawy WK, Abdel-Fattah YR, Soliman NA, Agblevor FA (2011) Comparison of alkaline pulping with steam explosion for glucose production from rice straw. Carbohydr Polym 83–2:720–726
Ishizawa CI, Davis MF, Schell DF, Johnson DK (2007) Porosity and its effect on the digestibility of dilute sulfuric acid pre-treated corn stover. J Agric Food Chem 55–7:2575–2581
Kim JS, Lee YY, Kim TH (2016) A review on alkaline pre-treatment technology for bioconversion of lignocellulosic biomass. Bioresour Technol 199:42–48
Kuhad RC, Gupta R, Khasa YP, Singh A (2010) Bioethanol production from Lantana camara (red sage): pre-treatment, saccharification and fermentation. Bioresour Technol 101–21:8348–8354
Kumar P, Barrett DM, Delwiche MJ, Stroeve P (2009) Methods for pre-treatment of lignocellulosic biomass for efficient hydrolysis and biofuel production. Ind Eng Chem 48–8:3713–3729
Mosier N, Wyman C, Dale B, Elander R, Lee YY, Holtzapple M, Ladisch M (2005) Features of promising technologies for pre-treatment of lignocellulosic biomass. Bioresour Technol 96–6:673–686
Moutta RO, Chandel AK, Rodrigues R, Silva MB, Rocha GJM, Silva SS (2012) Statistical optimization of sugarcane leaves hydrolysis into simple sugars by dilute sulfuric acid catalyzed process. Sugar Technol 14–1:53–60
Nelson N (1944) A Photometric adaptation of the somogyi method for the determination of glucose. J Biol Chem 153–2:375–380
Pasha C, Nagavalli M, Venkateswar Rao L (2007) Lantana camara for fuel ethanol production using thermotolerant yeast. Lett Appl Microbiol 44–6:666–672
Ranjan A, Moholkar VS (2013) Comparative study of various pre-treatment techniques for rice straw saccharification for the production of alcoholic biofuels. Fuel 112:567–571
Samuel R, Pu Y, Foston M, Ragauskas AJ (2010) Solid-state NMR characterization of switchgrass cellulose after dilute acid pretreatment. Biofuels 1–1:85–90
Segal L, Creely J, Martin JA, Conrad C (1962) An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer. Text Res J 29:786–794
Serrano-Ruiz JC, Ramos-Fernández EV, Sepúlveda-Escribano A (2012) From biodiesel and bioethanol to liquid hydrocarbon fuels: new hydrotreating and advanced microbial technologies. Energy Environ Sci 5–2:5638–5652
Sharma K, Kuhar S, Kuhad R, Bhat P (2007) Combinatorial approaches to improve plant cell wall digestion: possible solution for cattle feed problems. IK International Publishing House Pvt. Ltd., New Delhi
Sindhu R, Kuttiraja M, Binod P, Sukumaran RK, Pandey A (2014) Physicochemical characterization of alkali pre-treated sugarcane tops and optimization of enzymatic saccharification using response surface methodology. Renew Energy 62:362–368
Singh S, Moholkar VS, Goyal A (2014a) Optimization of carboxymethylcellulase production from Bacillus amyloliquefaciens SS35. 3Biotech 4(4):411–424
Singh S, Khanna S, Moholkar VS, Goyal A (2014b) Screening and optimization of pre-treatments for Parthenium hysterophorus as feedstock for alcoholic biofuels. Appl Energy 129:195–206
Somogyi MA (1945) New reagent for the determination of sugars. J Biol Chem 160:61–68
Sun Y, Cheng J (2002) Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresour Technol 83–1:1–11
TAPPI (1991) Technical Association of Pulp and Paper Industry. TAPPI Press, Atlanta, p 2
Zhao Y, Wang Y, Zhu JY, Ragauskas A, Deng Y (2008) Enhanced enzymatic hydrolysis of spruce by alkaline pre-treatment at low temperature. Biotechnol Bioeng 99–6:1320–1328
Zhu Z, Sathitsuksanoh N, Vinzant T, Schell DJ, McMillan JD, Zhang YHP (2009) Comparative study of corn stover pre-treated by dilute acid and cellulose solvent based lignocellulose fractionation: enzymatic hydrolysis, supermolecular structure, and substrate accessibility. Biotechnol Bioeng 103–4:715–724
The research work was supported by Centre for Bioenergy DBT-Pan-IIT Grant (BT/EB/PAN-IIT 2012) by the Department of Biotechnology, Ministry of Science and Technology, New Delhi, India. The authors acknowledge the use of FTIR spectrophotometer procured through the Indo-Finnish project Grant (BT/IN/Finland/08/AG/2011) from Department of Biotechnology (DBT), Ministry of Science and Technology, Government of India. The authors also acknowledge the Central Instrument Facility (CIF) at Indian Institute of Technology (IIT), Guwahati, for the provision of FESEM facilities. The authors also thankful to Department of Physics for providing XRD and Department of Biosciences and Bioengineering, IIT Guwahati for HPLC facility.
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Kumar, A., Singh, S., Rajulapati, V. et al. Evaluation of pre-treatment methods for Lantana camara stem for enhanced enzymatic saccharification. 3 Biotech 10, 37 (2020). https://doi.org/10.1007/s13205-019-2029-5
- Lantana camara stem
- Enzymatic saccharification