Insight into biodegradation of cellulose by psychrotrophic bacterium Pseudomonas sp. LKR-1 from the cold region of China: optimization of cold-active cellulase production and the associated degradation pathways

  • 60 Accesses

  • 1 Citations


In the cold regions of China, many lignocellulose-rich agricultural residues, such as corn stover, cannot be efficiently degraded due to low temperature. As the component of cellulose in corn stover was approximately 50%, the degradation of cellulose was considered as one of the most important processes in degrading corn stover. In this work, a psychrotrophic bacterial strain was screened from the soil in the cold region of China and identified as a Pseudomonas sp. named LKR-1, which is able to produce cold-active cellulase. To improve cellulase production, the fermentation conditions were optimized using the Box–Behnken design of the response surface methodology. The maximum cellulase activity was observed after 4 days of incubation at 13.7 °C and pH 7.6 with 8.18 g/L cellulose. HPLC, GC–MS and FTIR spectroscopy were used to describe the changes in residual cellulose and the products of cellulose degradation. In addition to glucose and cellobiose, 24 kinds of compounds were detected during cellulose degradation by psychrotrophic Pseudomonas sp. LKR-1. Furthermore, on the basis the data and references, the possible pathways of degradation by the psychrotrophic strain LKR-1 were speculated, which will help clarify and explain the mechanism of cellulose degradation by psychrotrophic bacteria. This result will provide valuable information that contributes to the exploration of the microbial degradation of corn stover polysaccharides in cold areas.

Graphic abstract

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 199

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Adav SS, Ng CS, Sze SK (2011) iTRAQ-based quantitative proteomic analysis of Thermobifida fusca reveals metabolic pathways of cellulose utilization. J. Proteom 74(10):2112–2122.

  2. Blanco-Canqui H, Lal R (2009) Crop residue removal impacts on soil productivity and environmental quality. Crit Rev Plant Sci 28(3):139–163.

  3. 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.

  4. Bugg TD, Ahmad M, Hardiman EM, Rahmanpour R (2011) Pathways for degradation of lignin in bacteria and fungi. Nat Prod Rep 28(12):1883–1896.

  5. Croce S, Wei Q, D’Imporzano G, Dong R, Adan 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(8):1289–1304.

  6. Dai Y, Yan Z, Jia L, Zhang S, Gao L, Wei X, Mei Z, Liu X (2016) The composition, localization and function of low-temperature-adapted microbial communities involved in methanogenic degradations of cellulose and chitin from Qinghai–Tibetan Plateau wetland soils. J Appl Microbiol 121:163–176.

  7. Demain AL, Newcomb M, Wu JH (2005) Cellulase, clostridia, and ethanol. Microbiol Mol Biol Rev 69(1):124–154.

  8. Deng TY, Liu HC (2014) Direct conversion of cellulose into acetol on bimetallic Ni-SnOx/Al2O3 catalysts. J Mol Catal A Chem 388–389:66–73.

  9. Dinis MJ, Bezerra RM, Nunes F, Dias AA, Guedes CV, Ferreira LM, Cone JW, Marques GS, Barros AR, Rodrigues MA (2009) Modification of wheat straw lignin by solid state fermentation with white-rot fungi. Bioresour Technol 100(20):4829–4835.

  10. Dupont AL (1996) Degradation of cellulose at the wet/dry interface. II. An approach to the identification of the oxidation compounds. Restaurator 17:145–164.

  11. Elanthikkal S, Gopalakrishnapanicker U, Varghese S, Guthrie JT (2010) Cellulose microfibres produced from banana plant wastes: isolation and characterization. Carbohydr Polym 80:852–859.

  12. Gangadharan D, Sivaramakrishnan S, Nampoothiri KM, Sukumaran RK, Pandey A (2008) Response surface methodology for the optimization of alpha amylase production by Bacillus amyloliquefaciens. Bioresour Technol 99(11):4597–4602.

  13. Gao DH, Chundawat SP, Sethi A, Balan V, Gnanakaran S, Dale BE (2013) Increased enzyme binding to substrate is not necessary for more efficient cellulose hydrolysis. Proc Natl Acad Sci USA 110(27):10922–10927.

  14. Garg P, Gupta A, Satya S (2006) Vermicomposting of different types of waste using Eisenia foetida: a comparative study. Bioresour Technol 97(3):391–395.

  15. Garrity GM, Bell JA, Lilburn TG (2004) Taxonomic outline of the prokaryotes. Bergey’s manual of systematic bacteriology. Seconded. Springer, New York

  16. Gehmayr V, Sixta H (2012) Pulp properties and their influence on enzymatic degradability. Biomacromolecules 13(3):645–651.

  17. Gupta P, Samant K, Sahu A (2012) Isolation of cellulose-degrading bacteria and determination of their cellulolytic potential. Int J Microbiol 2012:578925.

  18. 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 21:10386–10398.

  19. Jeong WS, Seo DH, Jung JH, Jung DH, Lee DW, Park YS, Park C (2017) Enzymatic characteristics of a highly thermostable beta-(1-4)-glucanase from Fervidobacterium islandicum AW-1 (KCTC 4680). J Microbiol Biotechnol 27(2):271–276.

  20. Ji N, Zhang T, Zheng M, Wang A, Wang H, Wang X, Chen JG (2008) Direct catalytic conversion of cellulose into ethylene glycol using nickel-promoted tungsten carbide catalysts. Angew Chem Int Ed 47(44):8510–8513.

  21. Kato K, Miura N (2008) Effect of matured compost as a bulking and inoculating agent on the microbial community and maturity of cattle manure compost. Bioresour Technol 99(9):3372–3380.

  22. Lattuati-Derieux A, Bonnassies-Termes S, Lavédrine B (2006) Characterisation of compounds emitted during natural and artificial ageing of a book. Use of headspace-solid-phase microextraction/gas chromatography/mass spectrometry. J Cult Herit 7:123–133.

  23. Lavoine N, Desloges I, Dufresne A, Bras J (2012) Microfibrillated cellulose—its barrier properties and applications in cellulosic materials: a review. Carbohydr Polym 90(2):735–764.

  24. Li L, Li K, Wang K, Chen C, Gao C, Ma C, Xu P (2014) Efficient production of 2,3-butanediol from corn stover hydrolysate by using a thermophilic Bacillus licheniformis strain. Bioresour Technol 170:256–261.

  25. Li HY, Li SN, Wang SX, Wang Q, Xue YY, Zhu BC (2015a) Degradation of lignocellulose in the corn straw by Bacillus amyloliquefaciens MN-8 (Chinese). Ying Yong Sheng Tai Xue Bao 26(5):1404–1410

  26. Li YH, Bai YX, Pan CM, Li WW, Zheng HQ, Zhang JN, Fan YT, Hou HW (2015b) Effective conversion of maize straw wastes into bio-hydrogen by two-stage process integrating H2 fermentation and MECs. Environ Sci Pollut Res Int 22(23):18394–18403.

  27. Li D, Feng L, Liu K, Cheng Y, Hou N, Li CY (2016) Optimization of cold-active CMCase production by psychrotrophic Sphingomonas sp. FLX-7 from the cold region of China. Cellulose 23(2):1335–1347.

  28. Liang YS, Yuan XZ, Zeng GM, Hu CL, Zhong H, Huang DL, Tang L, Zhao JJ (2010) Biodelignification of rice straw by Phanerochaete chrysosporium in the presence of dirhamnolipid. Biodegradation 21(4):615–624.

  29. Lin YC, Cho J, Tompsett GA, Westmoreland PR, Huber GW (2009) Kinetics and mechanism of cellulose pyrolysis. J Phys Chem C 113(46):20097–20107.

  30. Liu CF, Xu F, Sun JX, Ren JL, Curling S, Sun RC, Fowler P, Baird MS (2006) Physicochemical characterization of cellulose from perennial ryegrass leaves (Lolium perenne). Carbohydr Res 341(6):2677–2687.

  31. Liu D, Li J, Zhao S, Zhang R, Wang M, Miao Y, Shen Y, Shen Q (2013) Secretome diversity and quantitative analysis of cellulolytic Aspergillus fumigatus Z5 in the presence of different carbon sources. Biotechnol Biofuels 6(1):149.

  32. Lou H, Zhu JY, Lan TQ, Lai H, Qiu X (2013) pH-Induced lignin surface modification to reduce nonspecific cellulase binding and enhance enzymatic saccharification of lignocelluloses. ChemSusChem 6(5):919–927.

  33. Lü YC, Li N, Gong DC, Wang XF, Cui ZJ (2012) The effect of temperature on the structure and function of a cellulose-degrading microbial community. Appl Biochem Biotechnol 168(2):219–233.

  34. Lü Y, Li N, Yuan X, Hua B, Wang J, Ishii M, Igarashi Y, Cui Z (2013) Enhancing the cellulose-degrading activity of cellulolytic bacteria CTL-6 (Clostridium thermocellum) by co-culture with non-cellulolytic bacteria W2-10 (Geobacillus sp.). Appl Biochem Biotechnol 171(7):1578–1588.

  35. Marín M, Sánchez A, Artola A (2019) Production and recovery of cellulases through solid-state fermentation of selected lignocellulosic wastes. J Clean Prod 209:937–946.

  36. Mattonai M, Tamburini D, Colombini MP, Ribechini E (2016) Timing in analytical pyrolysis: Py(HMDS)-GC/MS of glucose and cellulose using online micro reaction sampler. Anal Chem 88(18):9318–9325.

  37. Melo IS, Zucchi TD, Silva RE, Vilela ES, Saber ML, Rosa LH, Pellizari VH (2014) Isolation and characterization of cellulolytic bacteria from the Stain house Lake, Antarctica. Folia Microbiol 59(4):303–306.

  38. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31(3):426–428.

  39. Nicolle J, Desauziers V, Mocho P, Ramalho O (2009) Optimization of FLEC-SPME for field passive sampling of VOCs emitted from solid building materials. Talanta 80:730–737.

  40. Patwardhan PR, Satrio JA, Brown RC, Shanks BH (2009) Product distribution from fast pyrolysis of glucose-based carbohydrates. J Anal Appl Pyrolysis 86(2):323–330.

  41. Qing GE, Gao JL, Yu XF, Zhang BL, Wang ZG, Borjigin NGCL, Hu SP, Sun JY, Xie M, Wang Z (2016) Screening of a microbial consortium with efifcient corn stover degradation ability at low temperature. J Integr Agric 15(10):2369–2379.

  42. Qiu XY, Hu SW (2013) “Smart” materials based on cellulose: a review of the preparations, properties, and applications. Materials (Basel) 6(3):738–781.

  43. Qu G, He W, Cai Y, Huang X, Ning P (2016) Catalytic pyrolysis of cellulose in ionic liquid [bmim]OTf. Carbohydr Polym 148:390–396.

  44. Qua EH, Hornsby PR, Sharma HSS, Lyons G (2011) Preparation and characterisation of cellulose nanofibres. J Mater Sci 46(18):6029–6045.

  45. Rostami S, Azhdarpoor A, Rostami M, Samaei MR (2016) The effects of simultaneous application of plant growth regulators and bioaugmentation on improvement of phytoremediation of pyrene contaminated soils. Chemosphere 161:219–223.

  46. Ruan Z, Shan Z, Jiang S, Lei S, Yi Z, Wang Y, Chao C, Zhao B (2013) Isolation and characterization of a novel cinosulfuron degrading Kurthia sp. from a methanogenic microbial consortium. Bioresour Technol 147:477–483.

  47. Sainsbury PD, Hardiman EM, Ahmad M, Otani H, Seghezzi N, Eltis LD, Bugg TDH (2013) Breaking down lignin to high-value chemicals: the conversion of lignocellulose to vanillin in a gene deletion mutant of Rhodococcus jostii RHA. ACS Chem Biol 8(10):2151–2156.

  48. Sanders EB, Goldsmith AI, Seeman JI (2003) A model that distinguishes the pyrolysis of -glucose, -fructose, and sucrose from that of cellulose. Application to the understanding of cigarette smoke formation. J Anal Appl Pyrol 66(1):29–50.

  49. Schedl A, Korntner P, Zweckmair T, Henniges U, Rosenau T, Potthast A (2016) Detection of cellulose-derived chromophores by ambient ionization-MS. Anal Chem 88(2):1253–1258.

  50. Schloss PD, Handelsman J (2005) Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71(3):1501–1506.

  51. Soares FL, Melo IS, Dias ACF, Andreote FD (2012) Cellulolytic bacteria from soils in harsh environments. World J Microbiol Biotechnol 28(5):2195–2203.

  52. 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.

  53. Sun RC, Tomkinson J, Ma PL, Liang SF (2000) Comparative study of hemicelluloses from rice straw by alkali and hydrogen peroxide treatments. Carbohydr Polym 42(2):111–122.

  54. Wang RF, Zhang JW, Dong ST, Liu P (2011) Present situation of maize straw resource utilization and its effect in main maize production regions of China (Chinese). Ying Yong Sheng Tai Xue Bao 22(6):1504–1510

  55. Wang S, Guo X, Liang T, Zhou Y, Luo Z (2012) Mechanism research on cellulose pyrolysis by Py-GC/MS and subsequent density functional theory studies. Bioresour Technol 104:722–728.

  56. Wang J, Wang X, Xu M, Feng G, Zhang W, Lu C (2015a) Crop yield and soil organic matter after long-term straw return to soil in China. Nutr Cycl Agroecosyst 102(3):371–381.

  57. 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.

  58. Wei TY, Zhang SQ, Shao LG, You YJ (2004) Isolation and Study of a new strain of cellulose degrading bacterium (Chinese). Environ Sci Technol 27(1–2):39

  59. Yang TH, Rathnasingh C, Lee HJ, Seung D (2014) Identification of acetoin reductases involved in 2,3-butanediol pathway in Klebsiella oxytoca. J Biotechnol 172:59–66.

  60. Zang HL, Yu Q, Lv TY, Cheng Y, Feng L, Cheng XS, Li CY (2016) Insights into the degradation of chlorimuron-ethyl by Stenotrophomonas maltophilia D310-3. Chemosphere 144:176–184.

Download references


This work was supported by the National Natural Science Foundation of China (No. 41771559). 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.

Author information

Correspondence to Chunyan Li.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 70 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Sun, S., Zhang, Y., Liu, K. et al. Insight into biodegradation of cellulose by psychrotrophic bacterium Pseudomonas sp. LKR-1 from the cold region of China: optimization of cold-active cellulase production and the associated degradation pathways. Cellulose 27, 315–333 (2020).

Download citation


  • Psychrotrophic bacterium
  • Pseudomonas sp.
  • Cellulose degradation
  • Cold-active cellulase
  • Degradation pathway