, Volume 16, Issue 6, pp 1033–1045 | Cite as

Effect of different additives on bacterial cellulose production by Acetobacter xylinum and analysis of material property

  • Kuan-Chen Cheng
  • Jeffrey M. CatchmarkEmail author
  • Ali Demirci


Bacterial cellulose (BC) demonstrates unique properties including high mechanical strength, high crystallinity, and high water retention ability, which make it an useful material in many industries, such as food, paper manufacturing, and pharmaceutical application. In this study, different additives including agar, carboxymethylcellulose (CMC), microcrystalline cellulose, and sodium alginate were added into fermentation medium in agitated culture to enhance BC production by Acetobacter xylinum. The optimal additive was chosen based on the amount of BC produced. The produced BC was analyzed by using X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), thermogravimetric analysis (TGA), and dynamic mechanical analysis (DMA). Among the evaluated additives, CMC yielded highest BC production (8.2 g/L) compared to the control (1.3 g/L). The results also indicated that CMC-altered BC production increased with CMC addition and reached saturation around 1%. The variation between replicates for all analysis was <5%. From XRD analysis, however, the crystallinity and crystal size decreased as CMC addition increased. FESEM results showed CMC-altered BC produced from agitated culture retained its interweaving property. TGA results demonstrated that CMC-altered BC had about 98% water retention ability, which is higher than BC pellicle produced with static culture. CMC-altered BC also exhibited higher Tmax compared to control. Finally, DMA results showed that BC from agitated culture loses its mechanical strength in both stress at break and Young’s modulus when compared to BC pellicle. This study clearly demonstrated that addition of CMC enhanced BC production and slightly changed its structure.


Bacterial cellulose Acetobacter xylinum Cellulose crystallinity, cellulose yield 



This work was supported in part by a seed grant from the College of Agricultural Sciences at the Pennsylvania State University and the Pennsylvania Experiment Station. The authors are very grateful to Nichole Wonderling from the Materials Research Institute of the Pennsylvania State University for her assistance with X-ray diffraction measurements. Thanks also to Yang Hu and Yufan Zeng in the Agricultural and Biological Engineering and Forest Resources department for useful discussion of DMA and TGA analysis.


  1. Astley OM, Chanliaud E, Donald AM et al (2003) Tensile deformation of bacterial cellulose composites. Int J Biol Macromol 32:28–35CrossRefGoogle Scholar
  2. Backdahl H, Helenius G, Bodin A et al (2006) Mechanical propertied of bacterial cellulose and interactions with smooth muscle cells. Biomaterials 27:2141–2149CrossRefGoogle Scholar
  3. Bae S, Sugano Y, Shoda M (2004) Improvement of bacterial cellulose production by addition of agar in a jar fermentor. J Biosci Bioeng 97(1):33–38CrossRefGoogle Scholar
  4. Benziman M, Haigler C, Brown RM et al (1980) Cellulose biogenesis: polymerization and crystallization are coupled process in Acetobacter xylinum. Proc Natl Acad Sci USA 77(11):6678–6682CrossRefGoogle Scholar
  5. Brown RM (2004) Cellulose structure and biosynthesis: what is in store for the 21th Century? J Poly Sci: Part A: Polymer Chem 42:487–495CrossRefGoogle Scholar
  6. Cook K, Colvin J (1980) Evidence for a beneficial influence of cellulose production on growth of Acetobacter xylinum in liquid medium. Current Microbiol 55:2448Google Scholar
  7. Czaja W, Romanovicz D, Brown RM (2004) Structural investigations of microbial cellulose produced in stationary and agitated culture. Cellulose 11:403–411CrossRefGoogle Scholar
  8. Delmer DP (1987) Cellulose biosynthesis. Annu Rev Plant Physiol 38:259–290CrossRefGoogle Scholar
  9. Gindl W, Keckes J (2004) Tensile properties of cellulose acetate butyrate composites reinforced with bacterial cellulose. Compos Sci Technol 64:2407–2413CrossRefGoogle Scholar
  10. Haigler CH, White AR, Brown RM Jr et al (1982) Alteration of in vivo cellulose ribbon assembly by carboxymethylcellulose and other cellulose derivatives. J Cell Biol 94(1):64–69CrossRefGoogle Scholar
  11. Hirai A, Tsuji M, Yamamoto H et al (1998) In situ crystallization of bacterial cellulose III. Influence of different polymeric additives on the formation of microfibrils as revealed by transmission electron microscopy. Cellulose 5:201–213CrossRefGoogle Scholar
  12. Hornung M, Ludwig M, Schmauder H-P (2007) Optimizing the production of bacterial cellulose in surface culture: a novel aerosol bioreactor working on a fed batch principle (part 3). Eng Life Sci 7:35–41CrossRefGoogle Scholar
  13. Hsieh YC, Yano H, Nogi M et al (2008) An estimation of the Young’s modulus of bacterial cellulose filaments. Cellulose 15:507–513CrossRefGoogle Scholar
  14. Hwang JW, Yang YK, Hwang JK et al (1999) Effects of pH and dissolved oxygen on cellulose production by Acetobacter xylinum BRC5 in agitated culture. J Biosci Bioeng 88(2):183–188CrossRefGoogle Scholar
  15. Ishida T, Mitarai M, Sugano Y et al (2003) Role of water-soluble polysaccharides in bacterial cellulose production. Biotechnol Bioeng 83(4):474–478CrossRefGoogle Scholar
  16. Ishikawa A, Matioka M, Tsuchida T et al (1995) Increasing of bacterial cellulose by sulfaguanidine-resistant mutants derived from Acetobacter xylinum subsp. sucrofermentants BPR 2001. Biosci Biotechnol Biochem 59:2259–2262CrossRefGoogle Scholar
  17. Kouda T, Yano H, Yoshinaga F (1997) Effect of agitator configuration on bacterial cellulose productivity in aerated and agitated culture. J Ferment Bioeng 83:371–376CrossRefGoogle Scholar
  18. Lynd LR, Weimer PJ, van Zyl WH et al (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66(3):506–577CrossRefGoogle Scholar
  19. Mihranyan A, Llagostera AP, Karmhag R et al (2004) Moisture sorption by cellulose powders of varying crystallinity. Int J Pharm 269:433–442CrossRefGoogle Scholar
  20. Nishi Y, Uryu M, Yamanaka S et al (1990) The structure and mechanical-properties of sheets prepared from bacterial cellulose. 2. Improvement of the mechanical-properties of sheets and their applicability to diaphragms of electroacoustic transducers. J Mater Sci 25:2997–3001CrossRefGoogle Scholar
  21. Seifert M, Hesse S, Kabrelian V et al (2003) Controlling the water content of never dried and reswollen bacterial cellulose by the addition of water-soluble polymers to the culture medium. J Poly Sci Part A Polymer Chemistry. 42:463–470CrossRefGoogle Scholar
  22. Serafica G, Mormino R, Bungay H (2002) Inclusion of solid partivle in bacterial cellulose. Appl Microbiol Biotechnol 58:756–760CrossRefGoogle Scholar
  23. Toyosaki H, Naritomi T, Seto A et al (1995) Screen of bacterial cellulose-producing Acetobacter strains suitable for agitated culture. Biosci Biotechnol Biochem 59:1498–1502CrossRefGoogle Scholar
  24. Tsioptsias C, Stefopoulos A, Kokkinomalis I et al (2008) Development of micro- and nano-porous composite materials by processing cellulose with ionic liquids and supercritical CO2. Green Chem 10:965–971CrossRefGoogle Scholar
  25. Um IC, Ki CS, Kweon HY et al (2004) Wet spinning of silk polymer II. Effect of drawing on the structure characteristics and properties of filament. Int J Biol Macromol 34:107–119CrossRefGoogle Scholar
  26. Watanabe K, Hori Y, Tabuchi M et al (1994) Structural features of bacterial cellulose vary depending on the cultural conditions. Proc Cellulose Spciety of Japan, Kyoto, pp 45–50Google Scholar
  27. Yamanaka S, Watanabe K, Kitamura N et al (1989) The structure and mechanical properties of sheets prepared from bacterial cellulose. J Mater Sci 24:3141–3145CrossRefGoogle Scholar
  28. Yang CM, Chen CY (2005) Synthesis, characterization and properties of polyanilines containing transition metal ions. Synth Met 153:133–136CrossRefGoogle Scholar
  29. Yoshino T, Asakura T, Toda K (1996) Cellulose Production by Acetobacter pasteurianus on silicon Membrane. J Ferment Bioeng 81(1):32–36CrossRefGoogle Scholar
  30. Zhang LN, Xi Q, Mo ZS et al (2003) Current researching methods on polymer physics. Wuhan College Press, Hubei, pp 194–195Google Scholar
  31. Zhou LL, Sun DP, Hu LY et al (2007) Effect of addition of sodium alginate on bacterial cellulose production by Acetobacter xylinum. J Ind Microbiol Biotechnol 34(7):483–489CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Kuan-Chen Cheng
    • 1
  • Jeffrey M. Catchmark
    • 1
    Email author
  • Ali Demirci
    • 1
    • 2
  1. 1.Department of Agricultural and Biological EngineeringThe Pennsylvania State UniversityUniversity ParkUSA
  2. 2.The Huck Institutes of Life SciencesThe Pennsylvania State UniversityUniversity ParkUSA

Personalised recommendations