pp 1–18 | Cite as

Biodegradation of lignin and the associated degradation pathway by psychrotrophic Arthrobacter sp. C2 from the cold region of China

  • Cheng Jiang
  • Yi Cheng
  • Hailian Zang
  • Xi Chen
  • Yue Wang
  • Yuting Zhang
  • Jinming Wang
  • Xiaohui Shen
  • Chunyan LiEmail author
Original Research


Degradation of most of the lignocellulose-rich agricultural residue in the cold regions of China is limited due to the cold climate. Lignin is the main component of lignocellulose, and the effective degradation of lignin is one of the most crucial processes in degrading lignocellulose. Psychrotrophic lignin-degrading bacteria and cold adapted ligninolytic enzymes have promising potential for the degradation and transformation of lignin, which are conducive to the resource utilization of lignocelluloses and energy-saving production under cold conditions. In this study, a newly psychrotrophic bacterial strain, Arthrobacter sp. C2, was isolated. The optimal enzyme activity conditions and lignin degradation pathways of C2 were investigated using sodium lignin sulfonate as substrate. The optimal conditions for enzyme activity included an initial pH of 6.74, a temperature of 14.9 °C, an incubation time of 6.87 days, and an inoculum size of 2.24%. Under the optimal conditions, the lignin peroxidase and manganese peroxidase activities and the degradation rate reached 29.8 U/L, 56.4 U/L and 40.1%, respectively. The biodegradation products including acids, phenols, aldehydes and alcohols were analyzed by gas chromatography–mass spectrometry and Fourier transform infrared spectroscopy. Further, the potential degradation pathways were proposed according to the results obtained in this study and those presented in the relevant literature. This study not only provides valuable psychrotrophic strain resources for the sustainable utilization of lignocellulose in cold regions, but also supplies potential application options for energy-saving production of useful chemicals using cold adapted enzymes.

Graphic abstract


Lignin Psychrotrophic bacterium Arthrobacter sp. Lignin peroxidase activity Manganese peroxidase activity Degradation pathway 



Lignin peroxidase


Manganese peroxidase


Gas chromatography–mass spectrometry


Fourier transform infrared spectroscopy


Response surface methodology


Box–Behnken design


Lignin mineral salt medium


Optical density


Retention time


Tricarboxylic acid



We would like to acknowledge “Northeast Agricultural University/Key Laboratory of Swine Facilities Engineering, Ministry of Agriculture, People’s Republic of China” for excellent technical assistance. This research was supported by the National Natural Science Foundation of China (Grant Numbers 41771559).

Authors’ contribution

CYL and HLZ designed the whole scheme of the study and conducted the experiments. CJ, YC, and XC performed experiments and XHS and JMW analyzed data. CJ and CYL wrote the manuscript, and YW and YTZ helped to revise. All authors read and approved the final manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethics approval and consent to participate

Not applicable.

Supplementary material

10570_2019_2858_MOESM1_ESM.docx (273 kb)
Supplementary file1 (DOCX 273 kb)


  1. Ahmad M, Roberts JN, Hardiman EM, Singh R, Eltis LD, Bugg TD (2011) Identification of DypB from Rhodococcus jostii RHA1 as a lignin peroxidase. Biochemistry 50:5096–5107. CrossRefPubMedGoogle Scholar
  2. Akila G, Chandra TS (2003) A novel cold-tolerant Clostridium strain PXYL1 isolated from a psychrophilic cattle manure digester that secretes thermolabile xylanase and cellulase. FEMS Microbiol Lett. 219:63–67. CrossRefPubMedGoogle Scholar
  3. Asina F, Brzonova I, Voeller K, Kozliak E, Kubátová A, Yao B, Ji Y (2016) Biodegradation of lignin by fungi, bacteria and laccases. Bioresour Technol. 220:414–424. CrossRefPubMedGoogle Scholar
  4. Bai Y-l, Wang L, Lu Y-l, Yang L-p, Zhou L-p, Ni L, Cheng M-f (2015) Effects of long-term full straw return on yield and potassium response in wheat-maize rotation. J Integr Agric. 14:2467–2476. CrossRefGoogle Scholar
  5. Blanco-Canqui H, Lal R (2009) Crop residue removal impacts on soil productivity and environmental quality. Crit Rev Plant Sci. 28:139–163. CrossRefGoogle Scholar
  6. Bloois EV, Pazmiño DET, Winter RT, Fraaije MW (2010) A robust and extracellular heme-containing peroxidase from Thermobifida fusca as prototype of a bacterial peroxidase superfamily. Appl Microbiol Biotechnol. 86:1419–1430. CrossRefPubMedGoogle Scholar
  7. Brodin M, Vallejos M, Opedal MT, Area MC, Chinga-Carrasco G (2017) Lignocellulosics as sustainable resources for production of bioplastics—a review. J Clean Prod. 162:646–664. CrossRefGoogle Scholar
  8. Brown ME, Barros T, Chang MC (2012) Identification and characterization of a multifunctional dye peroxidase from a lignin-reactive bacterium. ACS Chem Biol. 7:2074–2081. CrossRefPubMedGoogle Scholar
  9. Bugg TD, Ahmad M, Hardiman EM, Rahmanpour R (2011) Pathways for degradation of lignin in bacteria and fungi. Nat Prod Rep. 28:1883–1896. CrossRefPubMedGoogle Scholar
  10. Cao W, Xu H, Zhang H (2013) Architecture and functional groups of biofilms during composting with and without inoculation. Process Biochem. 48:1222–1226. CrossRefGoogle Scholar
  11. Chandra R, Bharagava RN (2013) Bacterial degradation of synthetic and kraft lignin by axenic and mixed culture and their metabolic products. J Environ Biol. 34:991–999. CrossRefPubMedGoogle Scholar
  12. Chen YH, Chai LY, Zhu YH, Yang ZH, Zheng Y, Zhang H (2012) Biodegradation of kraft lignin by a bacterial strain Comamonas sp. B-9 isolated from eroded bamboo slips. J Appl Microbiol. 112:900–906. CrossRefPubMedGoogle Scholar
  13. Croce S, Wei Q, D'Imporzano G, Dong R, Adani F (2016) Anaerobic digestion of straw and corn stover: the effect of biological process optimization and pre-treatment on total bio-methane yield and energy performance. Biotechnol Adv. 34:1289–1304. CrossRefPubMedGoogle Scholar
  14. Datta R, Kelkar A, Baraniya D, Molaei A, Moulick A, Meena R, Formanek P (2017) Enzymatic degradation of lignin in soil: a review. Sustainability. 9:1163. CrossRefGoogle Scholar
  15. Diao Y, Song M, Zhang Y, Shi LY, Lv Y, Ran R (2017) Enzymic degradation of hydroxyethyl cellulose and analysis of the substitution pattern along the polysaccharide chain. Carbohydr Polym. 169:92–100. CrossRefPubMedGoogle Scholar
  16. Dornez E, Verjans P, Arnaut F, Delcour JA, Courtin CM (2011) Use of psychrophilic xylanases provides insight into the xylanase functionality in bread making. J Agric Food Chem. 59:9553–9562. CrossRefPubMedGoogle Scholar
  17. Duan J, Liang JD, Du WJ, Wang DQ (2014) Biodegradation of kraft lignin by a bacterial strain Sphingobacterium sp. HY-H. Adv Mat Res. 955–959:548–553Google Scholar
  18. Fang S, An X, Liu H, Cheng Y, Hou N, Feng L, Huang X, Li C (2015) Enzymatic degradation of aliphatic nitriles by Rhodococcus rhodochrous BX2, a versatile nitrile-degrading bacterium. Bioresour Technol. 185:28–34. CrossRefPubMedGoogle Scholar
  19. Glenn JK, Akileswaran L, Gold MH (1986) Mn(II) oxidation is the principal function of the extracellular Mn-peroxidase from Phanerochaete chrysosporium. Arch Biochem Biophys. 251:688–696. CrossRefPubMedGoogle Scholar
  20. Guo X, Xie C, Wang L, Li Q, Wang Y (2019) Biodegradation of persistent environmental pollutants by Arthrobacter sp. Environ Sci Pollut Res Int. 26:8429–8443. CrossRefPubMedGoogle Scholar
  21. He T, Ye Q, Sun Q, Cai X, Ni J, Li Z, Xie D (2018) Removal of nitrate in simulated water at low temperature by a novel psychrotrophic and aerobic bacterium, Pseudomonas taiwanensis. Strain J Biomed Res Int 2018:4984087. CrossRefGoogle Scholar
  22. Hendriks AT, Zeeman G (2009) Pretreatments to enhance the digestibility of lignocellulosic biomass. Bioresour Technol. 100:10–18. CrossRefPubMedGoogle Scholar
  23. Hernandez-Pérez G, Goma G, Rols JL (1997) Degradation of lignosulfonated compounds by Streptomyces viridosporus strain T7A. Biotechnol Lett. 19:285–290. CrossRefGoogle Scholar
  24. Hildebrandt P, Wanarska M, Kur J (2009) A new cold-adapted beta-D-galactosidase from the Antarctic Arthrobacter sp. 32c—gene cloning, overexpression, purification and properties. BMC Microbiol. 9:151. Doi: 10.1186/1471-2180-9-151CrossRefGoogle Scholar
  25. Hong J, Ren L, Hong J, Xu C (2016) Environmental impact assessment of corn straw utilization in China. J Clean Prod. 112:1700–1708. CrossRefGoogle Scholar
  26. Horton RN, Apel WA, Thompson VS, Sheridan PP (2006) Low temperature reduction of hexavalent chromium by a microbial enrichment consortium and a novel strain of Arthrobacter aurescens. BMC Microbiol 6:1–8. CrossRefGoogle Scholar
  27. Hou N, Feng F, Shi Y, Cao H, Li C, Cao Z, Cheng Y (2014) Characterization of the extracellular biodemulsifiers secreted by Bacillus cereus LH-6 and the enhancement of demulsifying efficiency by optimizing the cultivation conditions. Environ Sci Pollut Res Int. 21:10386–10398. CrossRefPubMedGoogle Scholar
  28. Hou N, Wen L, Cao H, Liu K, An X, Li D, Wang H, Du X, Li C (2017) Role of psychrotrophic bacteria in organic domestic waste composting in cold regions of China. Bioresour Technol. 236:20–28. CrossRefPubMedGoogle Scholar
  29. Karimi M, Esfandiar R, Biria D (2017) Simultaneous delignification and saccharification of rice straw as a lignocellulosic biomass by immobilized Thrichoderma viride sp. to enhance enzymatic sugar production. Renew Energy. 104:88–95. CrossRefGoogle Scholar
  30. Kim SM, Park H, Choi JI (2016) Cloning and characterization of cold-adapted α-amylase from Antarctic Arthrobacter agilis. Appl Biochem Biotechnol. 181:1–12. CrossRefGoogle Scholar
  31. Kricka W, Fitzpatrick J, Bond U (2014) Metabolic engineering of yeasts by heterologous enzyme production for degradation of cellulose and hemicellulose from biomass: a perspective. Front Microbiol. 5:1–11. CrossRefGoogle Scholar
  32. Kumar L, Rathore V, Srivastava H (2001) 14C-[lignin]-lignocellulose biodegradation by bacteria isolated from polluted soil. Indian J Exp Biol. 39:584–589PubMedGoogle Scholar
  33. Li C, Zang H, Yu Q, Lv T, Cheng Y, Cheng X, Liu K, Liu W, Xu P, Lan C (2016a) Biodegradation of chlorimuron-ethyl and the associated degradation pathway by Rhodococcus sp. D310–1. Environ Sci Pollut Res Int. 23:8794–8805. CrossRefPubMedGoogle Scholar
  34. Li D, Feng L, Liu K, Cheng Y, Hou N, Li C (2016b) Optimization of cold-active CMCase production by psychrotrophic Sphingomonas sp. FLX-7 from the cold region of China. Cellulose 23:1335–1347. CrossRefGoogle Scholar
  35. Liang YL, Zhang Z, Wu M, Wu Y, Feng JX (2014) Isolation, screening, and identification of cellulolytic bacteria from natural reserves in the subtropical region of China and optimization of cellulase production by Paenibacillus terrae ME27-1. Biomed Res Int. 2014:1–13. CrossRefGoogle Scholar
  36. Liu Y, Hu T, Wu Z, Zeng G, Huang D, Shen Y, He X, Lai M, He Y (2014) Study on biodegradation process of lignin by FTIR and DSC. Environ Sci Pollut Res Int. 21:14004–14013. CrossRefPubMedGoogle Scholar
  37. Liu J, Sidhu SS, Ming LW, Tonnis B, Habteselassie M, Mao J, Huang Q (2015) Evaluation of various fungal pretreatment of switchgrass for enhanced saccharification and simultaneous enzyme production. J Clean Prod. 104:480–488. CrossRefGoogle Scholar
  38. Margesin R, Moertelmaier C, Mair J (2013) Low-temperature biodegradation of petroleum hydrocarbons (n-alkanes, phenol, anthracene, pyrene) by four actinobacterial strains. Int Biodeterior Biodegrad. 84:185–191. CrossRefGoogle Scholar
  39. Masai E, Katayama Y, Fukuda M (2007a) Genetic and biochemical investigations on bacterial catabolic pathways for lignin-derived aromatic compounds. J Agric Chem Soc Jpn. 71:1–15. CrossRefGoogle Scholar
  40. Masai E, Yamamoto Y, Inoue T, Takamura K, Hara H, Kasai D, Katayama Y, Fukuda M (2007b) Characterization of ligV essential for catabolism of vanillin by Sphingomonas paucimobilis SYK-6. Biosci Biotechnol Biochem. 71:2487. CrossRefPubMedGoogle Scholar
  41. Miranda-Ríos JA, Ramírez-Trujillo JA, Nova-Franco B, Lozano-Aguirre Beltrán LF, Iturriaga G, Suárez-Rodríguez R (2015) Draft genome sequence of Arthrobacter chlorophenolicus strain Mor30.16, isolated from the bean rhizosphere. Genome Announc. 3:1–2. CrossRefGoogle Scholar
  42. Mongodin EF, Shapir N, Daugherty SC, DeBoy RT, Emerson JB, Shvartzbeyn A, Radune D, Vamathevan J, Riggs F, Grinberg V, Khouri H, Wackett LP, Nelson KE, Sadowsky MJ (2006) Secrets of soil survival revealed by the genome sequence of Arthrobacter aurescens TC1. PLoS Genet. 2:e214. CrossRefPubMedPubMedCentralGoogle Scholar
  43. Niewerth H, Schuldes J, Parschat K, Kiefer P, Vorholt JA, Daniel R, Fetzner S (2012) Complete genome sequence and metabolic potential of the quinaldine-degrading bacterium Arthrobacter sp. Rue61a. Bmc Genom. 13:534–534. CrossRefGoogle Scholar
  44. Panagiotopoulos IA, Lignos GD, Bakker RR, Koukios EG (2012) Effect of low severity dilute-acid pretreatment of barley straw and decreased enzyme loading hydrolysis on the production of fermentable substrates and the release of inhibitory compounds. J Clean Prod. 32:45–51. CrossRefGoogle Scholar
  45. Perestelo F, Carnicero A, De lFG (1996) Short communication: isolation of a bacterium capable of limited degradation of industrial and labelled, natural and synthetic lignins. World J Microbiol Biotechnol. 12:111–112. CrossRefPubMedGoogle Scholar
  46. Priefert H, Rabenhorst J, Steinbüchel A (1997) Molecular characterization of genes of Pseudomonas sp. strain HR199 involved in bioconversion of vanillin to protocatechuate. J Bacteriol. 179:2595–2607. CrossRefPubMedPubMedCentralGoogle Scholar
  47. Raj A, Chandra R, Reddy MMK, Purohit HJ, Kapley A (2006) Biodegradation of kraft lignin by a newly isolated bacterial strain, Aneurinibacillus aneurinilyticus from the sludge of a pulp paper mill. World J Microbiol Biotechnol. 23:793–799. CrossRefGoogle Scholar
  48. Raj A, Krishna Reddy MM, Chandra R (2007) Identification of low molecular weight aromatic compounds by gas chromatography–mass spectrometry (GC–MS) from kraft lignin degradation by three Bacillus sp. Int Biodeter Biodegr. 59:292–296. CrossRefGoogle Scholar
  49. Ramachandra M, Crawford DL, Hertel G (1988) Characterization of an extracellular lignin peroxidase of the lignocellulolytic actinomycete Streptomyces viridosporus. Appl Environ Microbiol. 54:3057PubMedPubMedCentralGoogle Scholar
  50. Rouches E, Herpoëlgimbert I, Steyer JP, Carrere H (2016) Improvement of anaerobic degradation by white-rot fungi pretreatment of lignocellulosic biomass: a review. Renew Sustain Energy Rev. 59:179–198. CrossRefGoogle Scholar
  51. Ruan Z, Zhou S, Jiang S, Sun L, Zhai Y, Wang Y, Chen C, Zhao B (2013) Isolation and characterization of a novel cinosulfuron degrading Kurthia sp. from a methanogenic microbial consortium. Bioresour Technol. 147:477–483. CrossRefPubMedGoogle Scholar
  52. Sainsbury PD, Hardiman EM, Ahmad M, Otani H, Seghezzi N, Eltis LD, Bugg TD (2013) Breaking down lignin to high-value chemicals: the conversion of lignocellulose to vanillin in a gene deletion mutant of Rhodococcus jostii RHA1. ACS Chem Biol. 8:2151–2156. CrossRefPubMedGoogle Scholar
  53. Sathish L, Pavithra N, Ananda K (2012) Antimicrobial activity and biodegrading enzymes of endophytic fungi from eucalyptus. Int J Pharm Sci Res. 3:2574–2583Google Scholar
  54. See-Too WS, Ee R, Lim YL, Convey P, Pearce DA, Mohidin TBM, Yin WF, Chan KG (2017) Complete genome of Arthrobacter alpinus strain R3.8, bioremediation potential unraveled with genomic analysis. Stand Genomic Sci. 12:52. doi:10.1186/s40793–017–0264–0Google Scholar
  55. Shi Y, Chai L, Tang C, Yang Z, Zhang H, Chen R, Chen Y, Zheng Y (2013) Characterization and genomic analysis of kraft lignin biodegradation by the beta-proteobacterium Cupriavidus basilensis B-8. Biotechnol Biofuels. 6:1–1. CrossRefPubMedPubMedCentralGoogle Scholar
  56. Song J, Gu J, Yi Z, Wei W, Wang H, Ruan Z, Shi Y, Yan Y (2013) Biodegradation of nicosulfuron by a Talaromyces flavus LZM1. Bioresour Technol. 140:243–248. CrossRefPubMedGoogle Scholar
  57. Struvay C, Feller G (2012) Optimization to Low Temperature Activity in Psychrophilic Enzymes. Int J Mol Sci. 13:11643–11665. CrossRefPubMedPubMedCentralGoogle Scholar
  58. Sun Y, Qiu X, Liu Y (2013) Chemical reactivity of alkali lignin modified with laccase. Biomass Bioenergy. 55:198–204. CrossRefGoogle Scholar
  59. Tang M, Zhang F, Yao S, Liu Y, Chen J (2015) Application of Pseudomonas flava WD-3 for sewage treatment in constructed wetland in winter. Environ Technol. 36:1205–1211. CrossRefPubMedGoogle Scholar
  60. Tanvi CS, Dhanker R, Devi S, Goyal S (2018) Optimization of Physical and Nutritional Factors for Enhanced Production of Lignocellulolytic Enzymes by Aspergillus terreus FJAT-31011 under Submerged Conditions. Int J Curr Microbiol Appl Sci. 7:150–162. CrossRefGoogle Scholar
  61. Tien M, Kirk TK (1988) Lignin peroxidase of Phanerochaete chrysosporium. Methods Enzymol. 161:238–249. CrossRefGoogle Scholar
  62. Ueda M, Goto T, Nakazawa M, Miyatake K, Sakaguchi M, Inouye K (2010) A novel cold-adapted cellulase complex from Eisenia foetida: characterization of a multienzyme complex with carboxymethylcellulase, beta-glucosidase, beta-1,3 glucanase, and beta-xylosidase. Comp Biochem Physiol B Biochem Mol Biol. 157:26–32. CrossRefPubMedGoogle Scholar
  63. Wang D, Lin Y, Du W, Liang J, Ning Y (2013) Optimization and characterization of lignosulfonate biodegradation process by a bacterial strain Sphingobacterium sp. HY-H. Int Biodeterior Biodegrad. 85:365–371. CrossRefGoogle Scholar
  64. Wang J, Wang X, Xu M, Feng G, Zhang W, Ca Lu (2015a) Crop yield and soil organic matter after long-term straw return to soil in China. Nutr Cycl Agroecosys. 102:371–381. CrossRefGoogle Scholar
  65. Wang P, Chang J, Yin Q, Wang E, Zhu Q, Song A, Lu F (2015b) Effects of thermo-chemical pretreatment plus microbial fermentation and enzymatic hydrolysis on saccharification and lignocellulose degradation of corn straw. Bioresour Technol. 194:165–171. CrossRefPubMedGoogle Scholar
  66. Xiong M, Li C, Pan J, Cheng X, Xi C (2011) Isolation and characterization of Rhodococcus sp.BX2 capable of degrading bensulfuron-methyl. Afr J Microbiol Res. 5:4296–4302. CrossRefGoogle Scholar
  67. Yadav M, Paritosh K, Pareek N, Vivekanand V (2019) Coupled treatment of lignocellulosic agricultural residues for augmented biomethanation. J Clean Prod. 213:75–88. CrossRefGoogle Scholar
  68. Yoon JH, Oh HM, Yoon BD, Kang KH, Park YH (2003) Paenibacillus kribbensis sp. nov. and Paenibacillus terrae sp. nov., bioflocculants for efficient harvesting of algal cells. Int J Syst Evol Microbiol. 53:295–301. CrossRefPubMedGoogle Scholar
  69. Zang H, Yu Q, Lv T, Cheng Y, Feng L, Cheng X, Li C (2016) Insights into the degradation of chlorimuron-ethyl by Stenotrophomonas maltophilia D310–3. Chemosphere 144:176–184. CrossRefPubMedGoogle Scholar
  70. Zhang R, Zhou J, Gao Y, Guan Y, Li J, Tang X, Xu B, Ding J, Huang Z (2015) Molecular and biochemical characterizations of a new low-temperature active mannanase. Folia Microbiol. 60:483–492. CrossRefGoogle Scholar
  71. Zheng Y, Chai L-y, Yang Z-h, Zhang H, Chen Y-h (2013) Characterization of a newly isolated Bacterium Pandoraea sp. B-6 capable of degrading kraft lignin. J Cent South Univ. 20:757–763. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Cheng Jiang
    • 1
    • 2
  • Yi Cheng
    • 3
  • Hailian Zang
    • 1
  • Xi Chen
    • 1
  • Yue Wang
    • 1
  • Yuting Zhang
    • 1
  • Jinming Wang
    • 1
  • Xiaohui Shen
    • 4
  • Chunyan Li
    • 1
    Email author
  1. 1.College of Resource and EnvironmentNortheast Agricultural UniversityHarbinPeople’s Republic of China
  2. 2.College of Life ScienceJiamusi UniversityJiamusiPeople’s Republic of China
  3. 3.College of ScienceChina Agricultural UniversityBeijingPeople’s Republic of China
  4. 4.Jiamusi Branch of Heilongjiang Academy of Agricultural SciencesJiamusiPeople’s Republic of China

Personalised recommendations