Characterization of the lignin polymer in Brassicaceae family

Document Type: Original paper


1 Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran. Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

2 Department of Pharmacognosy, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.

3 Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.


Background and objectives: Residues of medicinal plants after extraction and weeds are suitable candidates for bioethanol production. Significant barriers exist to make the conversion of lignocellulosic feedstock to biofuel cost effective and environmentally friendly; one of which is the lignin polymer. Brassicaceae family is one of the potential targets for biofuel production. The structural characteristics of lignin from Hirschfeldia incana, Sisymbrium altissimum and Cardaria draba were studied in comparison to that of Brassica napus. Methods: Lignin deposition was observed by phloroglucinol and Mäule staining. The total lignin content was determined by Klason method. Maximum UV absorbance and FT-IR spectra were compared. Ratio of syringyl to guaiacyl lignin (S/G ratio) as a metric of lignin digestibility was determined by DFRC followed by GC-MS analysis. 1H-NMR spectra of the total lignin was compared with other spectroscopic methods. Results: Staining of thestem cross sections of C. draba showed higher G units in contrast to the higher S units in S. altissimum which was in agreement with 1H-NMR analysis. Total lignin content for H. incana, C. draba and S. altissimum was 27.10%, 23.8% and 24.5%, respectively. The specific maximum UV absorbance appeared between 230-260 nm. FT-IR analysis confirmed the presence of more aromatic structures in the seed maturation stage than the flowering stage. S/G ratio was 0.26, 0.10 and 0.22 for H. incana, C. draba and S. altissimum, respectively.  Conclusion: Except Cardaria draba with the predominance of G subunits in lignin polymer, Hirschfeldia incana and Sisymbrium altissimum are suitable candidates for bioethanol production.


[1] Li X, Weng JK, Chapple C. Improvement of biomass through lignin modification. Plant J. 2008; 54(4): 569-581.

[2] Simmons BA, Loque D, Blanch HW. Next-generation biomass feedstocks for biofuel production. Genome Biol. 2008; Article ID 19133109.

[3] Keating JD, Panganiban C, Mansfield SD. Tolerance and adaptation of ethanologenic yeasts to lignocellulosic inhibitory compounds. Biotechnol Bioeng. 2006; 93(6): 1196-1206.

[4] Agbor VB, Cicek N, Sparling R, Berlin A, Levin DB. Biomass pretreatment: fundamentals toward application. Biotechnol Adv. 2011; 29(6): 675-685.

[5] Wyman CE, Dale BE, Elander RT, Holtzapple M, Ladisch MR, Lee YY. Coordinated development of leading biomass pretreatment technologies. Bioresource Technol. 2005; 96(18): 1959-1966.

[6] Chang XF, Chandra R, Berleth T, Beatson RP. Rapid, microscale, acetyl bromide-based method for high-throughput determination of lignin content in Arabidopsis thaliana. J Agric Food Chem. 2008; 56(16): 6825-6834.

[7] Ralph J, Lundquist K, Brunow G, Lu F, Kim H, Schatz PF, Marita JM, Hatfield RD, Ralph SA, Christensen JH, Boerjan W. Lignins: Natural polymers from oxidative coupling of 4-hydroxyphenyl- propanoids. Phytochem Rev. 2004; 3(1): 29-60.

[8] Lupoi JS, Singh S, Davis M, Lee DJ, Shepherd M, Simmons BA, Henry RJ. High- syringyl/guaiacyl content using multivariate analysis a comparison between mid-infrared, near-infrared and Raman spectroscopies for model development. Bioenerg Res. 2014; Article ID24955114.

[9] Chen CL. Lignins occurance in woody tissues, isolation, reactions and structure. In: Lewin M, Goldstein IS, Eds. Wood structure and composition. New York: Marcel Dekker, 1991.

[10] Sun L, Varanasi P, Yang F, Loqué D, Simmons BA, Singh S. Rapid determination of syringyl: guaiacyl ratios using FT-Raman spectroscopy. Biotechnol Bioeng. 2012; 109(3): 647-656.

[11] Ragauskas AJ, Beckham GT, Biddy MJ, Chandra R, Chen F, Davis MF, Davison BH, Dixon RA, Gilna P, Keller M, Langan P, Naskar AK, Saddler JN, Tschaplinski TJ, Tuskan GA, Wyman CE. Lignin valorization: improving lignin processing in the biorefinery. Science.  2014; 344(6185): 710-719.

[12] Pederssetti MM, Palú F, da Silva EA, Rohling JH, Cardozo-Filho L, Dariva C. Extraction of canola seed (Brassica napus) oil using compressed propane and supercritical carbon dioxide. J Food Eng. 2011; 102(2): 189-196.

[13] Godfray HCJ, Beddington JR, Crute IR, Haddad L, Lawrence D,  Muir JF, Pretty J, Robinson S, Thomas SM, Toulmin C. Food security the challenge of feeding 9 billion people. Science. 2010; 327(5967): 812-818.

[14] Somerville C, Youngs H, Taylor C, Davis SC, Long SP. Feedstocks for lignocellulosic biofuels. Science. 2010; 329(5993): 790-792.

[15] Kamkar B, Dorri MA, Teixeira da Silva JA. Assessment of land suitability and the possibility and performance of a canola (Brassica napus L.) - soybean (Glycine max L.) rotation in four basins of Golestan province. Egypt J Remote Sens Space Sci. 2014; 17(1): 95-104.

[16] Kiaei M, Mahdavi S, Kialashaki A, Nemati M, Samariha A, Saghafi A. Chemical composition and morphological properties of canola plant and its potential application in pulp and paper industry. Cellulose Chem Technol. 2014; 48(1-2): 105-110.

[17] Couvreur TLP, Franzke A, Al-Shehbaz IA, Bakker FT, Koch MA, Mommenhof K. Molecular phylogenetics, temporal diversification, and principles of evolution in the mustard family (Brassicaceae). Mol Biol Evol. 2010; 27(1): 55-71.

[18] Darmency H, Fleury A. Mating system in Hirschfeldia incana and hybridization to oilseed rape. Weed Res. 2000; 40(2): 231-238.

[19] Lefol E, Danielou V, Darmency H, Boucher F, Maillet J, Renard M. Gene dispersal from transgenic crops. I. Growth of interspecific hybrids between oilseed rape and the wild hoary mustard. J Appl Ecol. 1995; 32(4): 803-805.

[20] Afsharypuor S, Jamali M. Volatile constituents of the flowering aerial parts, fruits and roots of Cardaria draba L. J Essent Oil Res. 2006; 18(6): 674-675.

[21] Mojab F, Kamalinejad M, Ghaderi N, Vahidipour HR. Phytochemical screening of some species of Iranian plants. Iran J Pharm Res. 2003; 2(2): 77-82.

[22] Radonić A, Blažević I, Mastelić J, Zekić M, Skočibušić M, Maravić A. Phytochemical analysis and antimicrobial activity of Cardaria draba (L.) Desv. volatiles. Chem Biodivers. 2011; 8(6): 1170-1181.

[23] Oraby HF, Ramadan MF. Impact of suppressing the caffeic acid O-methyltransferase (COMT) gene on lignin, fiber, and seed oil composition in Brassica napus transgenic plants. Eur Food Res Technol. 2015;240(5): 931-938.

[24] Bhinu VS, Li R, Huang J, Kaminskyj S, Sharpe A, Hannoufa A. Perturbation of lignin biosynthesis pathway in Brassica napus (canola) plants using RNAi. Can J Plant Sci. 2009; 89(3): 441-453.

[25] Pomar F, Merino F, Barceló AR. O-4-Linked coniferyl and sinapyl aldehydes in lignifying cell walls are the main targets of the Wiesner (phloroglucinol-HCl) reaction. Protoplasma. 2002; 220(1-2): 17-28.

[26] Weng JK, Akiyama T, Bonawitz ND, Li X, Ralph J, Chapple C. Convergent evolution of syringyl lignin biosynthesis via distinct pathways in the lycophyte Selaginella and flowering plants. Plant Cell. 2010; 22(4): 1033-1045

[27] Lu F, Ralph J. The DFRC method for lignin analysis. 2. monomers from isolated lignins. J Agric Food Chem. 1998; 46(2): 547-552.

[28] Bunzel M, Seiler A, Steinhart H. Characterization of dietary fiber lignins from fruits and vegetables using the DFRC method. J Agric Food Chem. 2005; 53(24): 9553-9559.

[29] Bunzel M, Schüssler A, Saha GT. Chemical characterization of klason lignin preparations from plant-based foods. J Agric Food Chem. 2011; 59(23): 12506-12513.

[30] Lu F, Ralph J. DFRC method for lignin analysis. 1. New method for β-aryl ether cleavage:  lignin model studies. J Agric Food Chem. 1997; 45(12): 4655-4660.

[31] Davin LB, Lewis NG. Lignin primary structures and dirigent sites. Curr Opin Biotech. 2005; 16(4): 407-415.

[32] Matsui N, Ohira T. Analysis of broad leaf lignin of Japanese angiospermous trees by DFRC (derivatization followed by reductive cleavage) method. Plant Physiol Biochem. 2013; 72: 112-115.

[33] Van Acker R, Vanholme R, Storme V, Mortimer JC, Dupree P, Boerjan W. Lignin biosynthesis perturbations affect secondary cell wall composition and saccharification yield in Arabidopsis thaliana. Biotechnol Biofuels. 2013; Article ID 23622268.

[34] Lourenco A, Neiva DM, Gominho J, Curt MD, Fernández J, Marques AV, Pereira H. Biomass production of four Cynara cardunculus clones and lignin composition analysis. Biomass Bioenerg. 2015; 76: 86-95.

[35] Kishimoto T, Chiba W, Saito K, Fukushima K, Uraki Y, Ubukata M. Influence of syringyl to guaiacyl ratio on the structure of natural and synthetic lignins. J Agric Food Chem. 2010; 58(2): 895-901.

[36] Sykes R, Yung M, Novaes E, Kirst M, Peter G, Davis M. High-throughput screening of plant cell-wall composition using pyrolysis molecular beam mass spectroscopy. In: Mielenz JR, Eds. Biofules: Methods and protocols, methods in molecular biology. New Jercy: Humana Press, 2009.

[37] Davison BH, Drescher SR, Tuskan GA, Davis MF, Nghiem NP. Variation of S/G ratio and lignin content in a Populus family influences the release of xylose by dilute acid hydrolysis. Appl Biochem Biotechnol. 2006; 130(1): 427-435.

[38] Huntley SK, Ellis D, Gilbert M, Chapple C, Mansfield SD. Significant increases in pulping efficiency in C4H-F5H-transformed poplars improved chemical savings and reduced environmental toxins.  J Agric Food Chem. 2003; 51(21): 6178-6183.

[39] Wiliam D, Moghaddam PR. In vitro digestibility and neutral detergent fiber and lignin contents of plant parts of nine forage species. J Agr Sci. 1998; 131(1): 51-58.

[40] George N, Yang Y, Wang Z, Sharma-Shivappa R, Tungate K. Suitability of canola residue for cellulosic ethanol   production.  Energy Fuels. 2010; 24(8): 4454-4458.

[41] Antrola AM, Lewis NG. Trends in lignin modification: a comprehensive analysis of the effects of genetic manipulations/mutations on lignification and vascular integrity. Phytochemistry. 2002; 61(3): 221-294.

[42] Li X, Ximenes E, Kim Y, Slininger M, Meilan R, Ladisch M, Chapple C. Lignin monomer composition affects Arabidopsis cell-wall degradability after liquid hot water pretreatment. Biotechnol Biofuels. 2010; Article ID 21126354.

[43] Jahan MS, Mun SP. Characteristics of dioxane lignins isolated at different ages of nalita wood (Trema orientalis).  J Wood Chem Technol. 2007; 27(2): 83-98.

[44] Abdullah HM, Abdul Latif MH, Attiya HG. Characterization and determination of lignin in different types of Iraqi Phoenix Date palm pruning woods. Int J Biol Macromol. 2013; 61: 340-346.