Applied Microbiology and Biotechnology

, Volume 97, Issue 14, pp 6189–6199 | Cite as

Metabolic flux analysis of Gluconacetobacter xylinus for bacterial cellulose production

  • Cheng Zhong
  • Gui-Cai Zhang
  • Miao Liu
  • Xin-Tong Zheng
  • Pei-Pei Han
  • Shi-Ru JiaEmail author
Biotechnological products and process engineering


Metabolic flux analysis was used to reveal the metabolic distributions in Gluconacetobacter xylinus (CGMCC no. 2955) cultured on different carbon sources. Compared with other sources, glucose, fructose, and glycerol could achieve much higher bacterial cellulose (BC) yields from G. xylinus (CGMCC no. 2955). The glycerol led to the highest BC production with a metabolic yield of 14.7 g/mol C, which was approximately 1.69-fold and 2.38-fold greater than that produced using fructose and glucose medium, respectively. The highest BC productivity from G. xylinus CGMCC 2955 was 5.97 g BC/L (dry weight) when using glycerol as the sole carbon source. Metabolic flux analysis for the central carbon metabolism revealed that about 47.96 % of glycerol was transformed into BC, while only 19.05 % of glucose and 24.78 % of fructose were transformed into BC. Instead, when glucose was used as the sole carbon source, 40.03 % of glucose was turned into the by-product gluconic acid. Compared with BC from glucose and fructose, BC from the glycerol medium showed the highest tensile strength at 83.5 MPa, with thinner fibers and lower porosity. As a main byproduct of biodiesel production, glycerol holds great potential to produce BC with superior mechanical and microstructural characteristics.


Bacterial cellulose Metabolic flux analysis Productivity Microstructure Crystallinity index 



The authors are grateful for the financial support from the National Natural Science Foundation of China (project no. 21106105, project no. 20976133), the Foundation of Tianjin Educational Committee (no. 20100602), and Changjiang Scholars and Innovative Research Team in University (no. IRT1166). We also gratefully acknowledge Rebecca G. Ong for her assistance in editing this manuscript.

Supplementary material

253_2013_4908_MOESM1_ESM.pdf (47 kb)
ESM 1 (PDF 46 kb)


  1. Bodin A, Backdahl H, Fink H, Gustafsson L, Risberg B, Gatenholm P (2006) Influence of cultivation conditions on mechanical and morphological properties of bacterial cellulose tubes. Biotechnol Bioeng 97:425–434CrossRefGoogle Scholar
  2. Braud HS (2007) Thermal characterization of bacterial cellulose-phosphate composite membranes. J Therm Anal Calorim 87:815–818CrossRefGoogle Scholar
  3. Cannon RE, Anderson SM (1991) Biogenesis of bacterial cellulose. Crit Rev Microbiol 17:435–447CrossRefGoogle Scholar
  4. Ha JH, Shah N, Ul-Islam M, Khan T, Park JK (2011) Bacterial cellulose production from a single sugar α-linked glucuronic acid-based oligosaccharide. Process Biochem 46:1717–1723CrossRefGoogle Scholar
  5. Heo MS, Son HJ (2002) Development of an optimized simple chemically defined medium for bacterial cellulose production by Acetobacter sp. A9 in shaking cultures. Biotechnol Appl Biochem 36:41–45CrossRefGoogle Scholar
  6. Huang D, Wen JP, Wang GY, Yu GH, Jia XQ, Chen YL (2012) In silico aided metabolic engineering of Streptomyces roseosporus for daptomycin yield improvement. Appl Microbiol Biotechnol 94:637–649CrossRefGoogle Scholar
  7. Li YJ, Tian CJ, Tian H, Zhang JL, He X, Ping WX, Lei H (2012) Improvement of bacterial cellulose production by manipulating the metabolic pathways in which ethanol and sodium citrate involved. Appl Microbiol Biotechnol 96:1479–1487CrossRefGoogle Scholar
  8. Jung HI, Jeong JH, Lee OM, Park GT, Kim KK, Park HC, Lee SM, Kim YG, Kim HG, Son HJ (2010) Influence of glycerol on production and structural–physical properties of cellulose from Acetobacter sp. V6 cultured in shake flasks. Bioresour Technol 101:3602–3608CrossRefGoogle Scholar
  9. Keshk S, Sameshima K (2006) The utilization of sugar cane molasses with/without the presence of lignosulfonate for the production of bacterial cellulose. Appl Microbiol Biotechnol 72:291–296CrossRefGoogle Scholar
  10. Klemm D, Heublein B, Fink HP, Bohn A (2005) Cellulose: fascinating biopolymer and sustainable raw material. Angew Chem Int Ed 44:3358–3393CrossRefGoogle Scholar
  11. Klemm D, Schumann D, Kramer F, Hessler N, Hornung M, Schmauder H, Marsch S (2006) Nanocelluloses as innovative polymers in research and application. Adv Polym Sci 205:49–96CrossRefGoogle Scholar
  12. Ma K, Zhao HX, Zhang C, Lu Y, Xing XH (2012) Impairment of NADH dehydrogenase for increased hydrogen production and its effect on metabolic flux redistribution in wild strain and mutants of Enterobacter aerogenes. Int J Hydrogen Energy. doi: 10.1016/j.ijhydene. 2012.08.017
  13. Matsuoka M, Tsuchida T, Matsushita K, Adachi O, Yoshinaga F (1996) A synthetic medium for bacterial cellulose production by Acetobacter xylinum subsp. sucrofermentans. Biosci Biotechnol Biochem 100:575–579CrossRefGoogle Scholar
  14. Mikkelsen D, Flanagan BM, Dykes GA, Gidley MJ (2009) Influence of different carbon sources on bacterial cellulose production by Gluconacetobacter xylinus strain ATCC 53524. J Appl Microbiol 107:576–583CrossRefGoogle Scholar
  15. Naritomi T, Kouda T, Yano H, Yoshinaga F (1998) Effect of lactate on bacterial cellulose production from fructose in continuous culture. J Ferm Bioeng 85:89–95Google Scholar
  16. Nguyen VY, Flanagan B, Gidley MJ, Dykes GA (2008) Characterization of cellulose production by a Gluconacetobacter xylinus strain from kombucha. Curr Microbiol 57:449–453CrossRefGoogle Scholar
  17. Oh SY, Yoo DI, Shin Y, Kim HC (2005) Crystalline structure analysis of cellulose treated with sodium hydroxide and carbon dioxide by means of X-ray diffraction and FTIR spectroscopy. Carbohydr Res 340:2376–2391CrossRefGoogle Scholar
  18. Oikawa T, Nakai J, Tsukagawa Y, Soda K (1997) A novel type of d-mannitol dehydrogenase from Acetobacter xylinum occurrence purification and basic properties. Biosci Biotechnol Biochem 61:1778–1782CrossRefGoogle Scholar
  19. Park ST, Kim E, Kim YM (2006) Overproduction of cellulose in Acetobacter xylinum KCCM 10100 defective in GDP-mannosyltransferase. J Microbiol Biotechnol 16:961–964Google Scholar
  20. Ross P, Mayer R, Benziman M (1991) Cellulose biosynthesis and function in bacteria. Microbiol Rev 55:35–58Google Scholar
  21. Ruka DR, Simon GP, Dean GM (2012) Altering the growth conditions of Gluconacetobacter xylinus to maximize the yield of bacterial cellulose. Carbohydr Polym 89:613–622CrossRefGoogle Scholar
  22. Saenge C, Cheirsilp B, Suksaroge TT, Bourtoom T (2011) Potential use of oleaginous red yeast Rhodotcorula glutinis for the bioconversion of crude glycerol from biodiesel plant to lipids and carotenoids. Process Biochem 46:210–218CrossRefGoogle Scholar
  23. Schaub J, Mauch K, Reuss M (2008) Metabolic flux analysis in Escherichia coli by integrating isotopic dynamic and isotopic stationary 13C labeling data. Biotechnol Bioeng 99:1170–1185CrossRefGoogle Scholar
  24. Schramm M, Gromet Z, Hestrin S (1957) Role of hexose phosphate in synthesis of cellulose by Acetobacter xylinum. Nature 179:28–29CrossRefGoogle Scholar
  25. Segal L, Creely J, Martin A, Conrad C (1959) An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer. Text Res J 29:786–794CrossRefGoogle Scholar
  26. Shezad O, Khan S, Khan T, Park JK (2010) Physicochemical and mechanical characterization of bacterial cellulose produced with an excellent productivity in static conditions using a simple fed-batch cultivation strategy. Carbohydr Polym 82:173–180CrossRefGoogle Scholar
  27. Shigematsu T, Takamine K, Kitazato M, Morita T, Naritomi T, Morimura S, Kida K (2005) Cellulose production from glucose using a glucose dehydrogenase gene (gdh)-deficient mutant of Gluconacetobacter xylinus and its use for bioconversion of sweet potato pulp. J Biosci Bioeng 99:415–422CrossRefGoogle Scholar
  28. Sturcová A, His I, Apperley DC, Sugiyama J, Jarvis MC (2004) Structural details of crystalline cellulose from higher plants. Biomacromolecules 5:1333–1339Google Scholar
  29. Svensson A, Nicklasson E, Harrah T, Panilaitis B, Kaplan DL, Brittberg M, Gatenholm P (2005) Bacterial cellulose as a potential scaffold for tissue engineering of cartilage. Biomaterials 26:419–431CrossRefGoogle Scholar
  30. Tamahkar E, Babac C, Kutsl T, Piskin E, Denizli A (2010) Bacterial cellulose nanofibers for albumin depletion from human serum. Process Biochem 45:1713–1719CrossRefGoogle Scholar
  31. Tang WH, Jia SR, Jia YY, Yang HJ (2010) The influence of fermentation conditions and post-treatment methods on porosity of bacterial cellulose membrane. World J Microbiol Biotechnol 26:125–131CrossRefGoogle Scholar
  32. Toda K, Asakura T, Fukaya M, Entani E, Kawamrua Y (1997) Cellulose production by acetic acid-resistant Acetobacter xylinum. J Ferment Bioeng 84:228–231CrossRefGoogle Scholar
  33. Tonouchi N, Sugiyama M, Yokozeki K (2003) Coenzyme specificity of enzymes in the oxidative pentose phosphate pathway of Gluconobacter oxydans. Biosci Biotechnol Biochem 67:2648–2651CrossRefGoogle Scholar
  34. UI-Islam M, Khan T, Park JK (2012) Nanoreinforced bacterial cellulose–montmorillonite composites for biomedical applications. Carbohyd Polym 89:1189–1197CrossRefGoogle Scholar
  35. Vandamme EJ, De Baets S, Vanbaelen A, Joris K, De Wulf P (1998) Improved production of bacterial cellulose and its application potential. Polym Degrad Stab 59:93–99CrossRefGoogle Scholar
  36. Velasco-Bedran H, Lopez-Isunza F (2007) The unified metabolism of Gluconacetobacter entanii in continuous and batch processes. Proc Biochem 42:1180–1190CrossRefGoogle Scholar
  37. Wan YZ, Hong L, Jia SR, Huang Y, Zhu Y, Wang YL, Jiang HL (2006) Synthesis and characterization of hydroxyapatite-bacterial cellulose nanocomposites. Compos Sci Technol 66:1825–1832CrossRefGoogle Scholar
  38. Weinhouse H, Benziman M (1976) Phosphorylation of glycerol and dihydroxyacetone in Acetobacter xylinum and its possible regulatory role. J Bacteriol 127:747–754Google Scholar
  39. Yunoki S, Osada Y, Kono H, Takai M (2004) Role of ethanol in improvement of bacterial cellulose production: analysis using 13C-labeled carbon sources. Food Sci Technol Res 10:307–313CrossRefGoogle Scholar
  40. Zeng XB, Small DP, Wan WK (2011) Statistical optimization of culture conditions for bacterial cellulose production by Acetobacter xylinum BPR 2001 from maple syrup. Carbohydr Polym 86:1558–1564CrossRefGoogle Scholar
  41. Zhong C, Cao XY, Li BZ, Yuan YJ (2009a) Biofuels in China: past, present and future. Biofuels Bioprod Bioref 3:247–270CrossRefGoogle Scholar
  42. Zhong C, Lau MW, Balan V, Dale BE, Yuan YJ (2009b) Optimization of enzymatic hydrolysis and ethanol fermentation from AFEX-treated rice straw. Appl Microbiol Biotechnol 84:667–676CrossRefGoogle Scholar
  43. Zhu HX, Jia SR, Wan T, Jia YY, Yang HJ, Li J, Yan L, Zhong C (2011) Biosynthesis of spherical Fe3O4/bacterial cellulose nanocomposites as adsorbents for heavy metal ions. Carbohydr Polym 86:1558–1564CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Cheng Zhong
    • 1
    • 2
    • 3
  • Gui-Cai Zhang
    • 1
    • 2
  • Miao Liu
    • 1
    • 2
  • Xin-Tong Zheng
    • 1
    • 2
  • Pei-Pei Han
    • 1
    • 2
  • Shi-Ru Jia
    • 1
    • 2
    Email author
  1. 1.Key Laboratory of Industrial Fermentation Microbiology (Ministry of Education)Tianjin University of Science and TechnologyTianjinPeople’s Republic of China
  2. 2.School of BiotechnologyTianjin University of Science and TechnologyTianjinPeople’s Republic of China
  3. 3.Key Laboratory of Systems Bioengineering, Ministry of EducationTianjin UniversityTianjinPeople’s Republic of China

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