, Volume 26, Issue 4, pp 2279–2290 | Cite as

DDA (degree of deacetylation) and pH-dependent antibacterial properties of chitin nanofibers against Escherichia coli

  • Junhua Xu
  • Liang Liu
  • Juan Yu
  • Yujun Zou
  • Zhiguo Wang
  • Yimin FanEmail author
Original Research


Partially deacetylated chitin nanofibers (PD-ChNFs) with different degrees of deacetylation (DDA: 16%, 29%, 38%, 47%) that represented different amino group contents were prepared by partial deacetylation treatment under the desired conditions and the subsequent mechanical treatment. Higher concentration of NaOH or treating for longer time will result a higher DDA. An equation for the calculation of the cationic amino group contents of PD-ChNFs was derived for the evaluation of the antibacterial effect of chitin nanofibers as a function of the DDA and pH. The amount of cationic amino groups was correlated with a higher DDA of PD-ChNFs and a lower pH. Furthermore, the antibacterial property of chitin nanofibers depended strongly on the pH and secondary on DDA that indicated better antibacterial properties were obtained with a higher amount of cationic amino groups. When the pH was higher than 7, there are almost no cationic amino groups and no obvious antibacterial effect was observed. For a pH lower than 5, almost all the amino groups were cationized and the growth of E. coli was effectively inhibited. And if pH was between 5 and 7, PD-ChNFs possessed higher DDA at lower pH carry more cationic amino group and show better antibacterial activity. Besides, the chitin nanofibers with higher amino group contents with a higher DDA prepared by more caustic deacetylation treatment tended to have smaller size, which might be another reason for the better antibacterial effect.

Graphical abstract


Chitin nanofiber Antibacterial Cationic amino group Degree of deacetylation 



This research was supported by the National Key R&D Program of China (2016YFD0600803) and the National Natural Science Foundation of China (No. 31100426). The authors also give acknowledgment to Advanced Analysis and Testing Center of Nanjing Forestry University.


  1. Aleanizy FS, Alqahtani FY, Shazly G et al (2018) Measurement and evaluation of the effects of pH gradients on the antimicrobial and antivirulence activities of chitosan nanoparticles in Pseudomonas aeruginosa. Saudi Pharm J 29(1):79–83CrossRefGoogle Scholar
  2. Benhabiles MS, Salah R, Lounici H et al (2012) Antibacterial activity of chitin, chitosan and its oligomers prepared from shrimp shell waste. Food Hydrocoll 29:48–56. CrossRefGoogle Scholar
  3. Chang SH, Lin HTV, Wu GJ, Tsai GJ (2015) pH Effects on solubility, zeta potential, and correlation between antibacterial activity and molecular weight of chitosan. Carbohydr Polym 134:74–81. CrossRefGoogle Scholar
  4. Chen CS, Liau WY, Tsai GJ (1998) Antibacterial effects of N-sulfonated and N-sulfobenzoyl chitosan and application to oyster preservation. J Food Prot 61:1124–1128. CrossRefGoogle Scholar
  5. Chien RC, Yen MT, Mau JL (2016) Antimicrobial and antitumor activities of chitosan from shiitake stipes, compared to commercial chitosan from crab shells. Carbohydr Polym 138:259–264. CrossRefGoogle Scholar
  6. Dutta AK, Egusa M, Kaminaka H et al (2015) Facile preparation of surface N-halamine chitin nanofiber to endow antibacterial and antifungal activities. Carbohydr Polym 115:342–347. CrossRefGoogle Scholar
  7. Fan Y, Saito T, Isogai A (2008) Chitin nanocrystals prepared by TEMPO-mediated oxidation of α-chitin. Biomacromol 9:192–198. CrossRefGoogle Scholar
  8. Fan Y, Saito T, Isogai A (2010) Individual chitin nano-whiskers prepared from partially deacetylated α-chitin by fibril surface cationization. Carbohydr Polym 79:1046–1051. CrossRefGoogle Scholar
  9. Goy RC, De Britto D, Assis OBG (2009) A review of the antimicrobial acitivity of chitosan. Polímeros Ciência e Tecnol 19:241–247. CrossRefGoogle Scholar
  10. Grant S, Blair HS, McKay G (1989) Structural studies on chitosan and other chitin derivatives. Die Makromol Chemie 190:2279–2286. CrossRefGoogle Scholar
  11. Hu H, Yu A, Kim E et al (2005) Influence of the zeta potential on the dispersability and purification of single-walled carbon nanotubes. J Phys Chem B 109:11520–11524. CrossRefGoogle Scholar
  12. Hwang KT, Jung ST, Lee GD et al (2002) Controlling molecular weight and degree of deacetylation of chitosan by response surface methodology. J Agric Food Chem 50:1876–1882. CrossRefGoogle Scholar
  13. Je JY, Kim SK (2006) Antimicrobial action of novel chitin derivative. Biochim Biophys Acta: Gen Subj 1760:104–109. CrossRefGoogle Scholar
  14. Jia R, Duan Y, Fang Q et al (2016) Pyridine-grafted chitosan derivative as an antifungal agent. Food Chem 196:381–387. CrossRefGoogle Scholar
  15. Jiang J, Ye W, Yu J, Fan Y, Ono Y, Saito T et al (2018a) Chitin nanocrystals prepared by oxidation of α-chitin using the o 2/laccase/tempo system. Carbohyd Polym 189:S014486171830122XCrossRefGoogle Scholar
  16. Jiang J, Ye W, Yu J, Fan Y, Ono Y, Saito T, Isogai A (2018b) Chitin nanocrystals prepared by oxidation of α-chitin using the O2/laccase/TEMPO system. Carbohyd Polym 189:178–183CrossRefGoogle Scholar
  17. Jung BO, Kim CH, Choi KS et al (1999) Preparation of amphiphilic chitosan and their antimicrobial activities. J Appl Polym Sci 72:1713–1719.;2-T CrossRefGoogle Scholar
  18. Jung EJ, Youn DK, Lee SH et al (2010) Antibacterial activity of chitosans with different degrees of deacetylation and viscosities. Int J Food Sci Technol 45:676–682. CrossRefGoogle Scholar
  19. Li J, Revol JF, Marchessault RH (1996a) Rheological properties of aqueous suspensions of chitin crystallites. J Colloid Interface Sci 183:365–373. CrossRefGoogle Scholar
  20. Li J, Revol JF, Naranjo E, Marchessault RH (1996b) Effect of electrostatic interaction on phase separation behaviour of chitin crystallite suspensions. Int J Biol Macromol 18:177–187. CrossRefGoogle Scholar
  21. Li J, Revol J-F, Marchessault RH (1997) Effect of degree of deacetylation of chitin on the properties of chitin crystallites. J Appl Polym 65:373–380.;2-0 CrossRefGoogle Scholar
  22. Li Z, Zhang M, Cheng D, Yang R (2016) Preparation of silver nano-particles immobilized onto chitin nano-crystals and their application to cellulose paper for imparting antimicrobial activity. Carbohydr Polym 151:834–840. CrossRefGoogle Scholar
  23. Li P, Zhao J, Chen Y et al (2017) Preparation and characterization of chitosan physical hydrogels with enhanced mechanical and antibacterial properties. Carbohydr Polym 157:1383–1392. CrossRefGoogle Scholar
  24. Liu H, Du Y, Wang X, Sun L (2004) Chitosan kills bacteria through cell membrane damage. Int J Food Microbiol 95:147–155. CrossRefGoogle Scholar
  25. Liu N, Sun Y, Zhou D (2015) Antimcriobial activity and mechanism of chitosan with different molecular weight. In: Proceedings—2015 7th international conference on measuring technology and mechatronics automation, ICMTMA 2015, pp 166–169Google Scholar
  26. Liu L, Wang R, Yu J et al (2016) Robust self-standing chitin nanofiber/nanowhisker hydrogels with designed surface charges and ultralow mass content via gas phase coagulation. Biomacromol 17:3773–3781. CrossRefGoogle Scholar
  27. Minke R, Blackwell J (1978) The structure of alpha-chitin. J Mol Biol 120:167–181. CrossRefGoogle Scholar
  28. Nguyen VQ, Ishihara M, Kinoda J et al (2014) Development of antimicrobial biomaterials produced from chitin-nanofiber sheet/silver nanoparticle composites. J Nanobiotechnol. Google Scholar
  29. Papineau AM, Hoover DG, Knorr D, Farkas DF (1991) Antimicrobial effect of water-soluble chitosans with high hydrostatic pressure. Food Biotechnol 5:45–57. CrossRefGoogle Scholar
  30. Pearson FG, Marchessault RH, Liang CY (1960) Infrared spectra of crystalline polysaccharides. V. Chitin. J Polym Sci 43:101–116. CrossRefGoogle Scholar
  31. Pereira AGB, Muniz EC, Lo Hsieh Y (2014) Chitosan-sheath and chitin-core nanowhiskers. Carbohydr Polym 107:158–166. CrossRefGoogle Scholar
  32. Pillai CKS, Paul W, Sharma CP (2009) Chitin and chitosan polymers: chemistry, solubility and fiber formation. Prog Polym Sci 34:641–678CrossRefGoogle Scholar
  33. Qi ZD, Fan Y, Saito T et al (2013) Improvement of nanofibrillation efficiency of α-chitin in water by selecting acid used for surface cationisation. RSC Adv 3:2613–2619. CrossRefGoogle Scholar
  34. Rinaudo M (2006) Chitin and chitosan: properties and applications. Prog Polym Sci 31:603–632CrossRefGoogle Scholar
  35. Shankar S, Reddy JP, Rhim JW, Kim HY (2015) Preparation, characterization, and antimicrobial activity of chitin nanofibrils reinforced carrageenan nanocomposite films. Carbohydr Polym 117:468–475. CrossRefGoogle Scholar
  36. Shigemasa Y, Matsuura H, Sashiwa H, Saimoto H (1996) Evaluation of different absorbance ratios from infrared spectroscopy for analyzing the degree of deacetylation in chitin. Int J Biol Macromol 18:237–242. CrossRefGoogle Scholar
  37. Takahashi T, Imai M, Suzuki I, Sawai J (2008) Growth inhibitory effect on bacteria of chitosan membranes regulated with deacetylation degree. Biochem Eng J 40:485–491. CrossRefGoogle Scholar
  38. Wang R, Liu L, Yu J et al (2017) Versatile protonic acid mediated preparation of partially deacetylated chitin nanofibers/nanowhiskers and their assembling of nano-structured hydro- and aero-gels. Cellulose 24:5443–5454. CrossRefGoogle Scholar
  39. Wu XY, Zeng QX, Mo SF, Ruan Z (2006) Antibacterial activities of chitosans with different Degrees of Deacetylation and Molecular Masses. Huanan Ligong Daxue Xuebao/J South China Univ Technol (Nat Sci) 34:58–62Google Scholar
  40. Ye W, Liu L, Yu J, Liu S, Yong Q, Fan Y (2018) Hypolipidemic activities of partially deacetylated α-chitin nanofibers/nanowhiskers in mice. Food Nutr Res 62:1295–1303CrossRefGoogle Scholar
  41. Zhang Y, Jiang J, Liu L et al (2015) Preparation, assessment, and comparison of α-chitin nano-fiber films with different surface charges. Nanoscale Res Lett 10:226. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Jiangsu Key Lab of Biomass-Based Green Fuel and Chemicals, College of Chemical EngineeringNanjing Forestry UniversityNanjingChina

Personalised recommendations