Abstract
Despite excellent biocompatibility and mechanical properties, the poor in vitro and in vivo degradability of cellulose has limited its biomedical and biomass conversion applications. To address this issue, we report a metabolic engineering-based approach to the rational redesign of cellular metabolites to introduce N-acetylglucosamine (GlcNAc) residues into cellulosic biopolymers during de novo synthesis from Gluconacetobacter xylinus. The cellulose produced from these engineered cells (modified bacterial cellulose [MBC]) was evaluated and compared with cellulose produced from normal cells (bacterial cellulose [BC]). High GlcNAc content and lower crystallinity in MBC compared to BC make this a multifunctional bioengineered polymer susceptible to lysozyme, an enzyme widespread in the human body, and to rapid hydrolysis by cellulase, an enzyme commonly used in biomass conversion. Degradability in vivo was demonstrated in subcutaneous implants in mice, where modified cellulose was completely degraded within 20 days. We provide a new route toward the production of a family of tailorable modified cellulosic biopolymers that overcome the longstanding limitation associated with the poor degradability of cellulose for a wide range of potential applications.
MeSH Terms
Analysis of Variance
Animals
Bioengineering/methods
Biopolymers/biosynthesis,metabolism
Cellulose/metabolism,ultrastructure
Chitin/metabolism
Chromatography, Liquid
Cloning, Molecular
DNA Primers/genetics
Electroporation
Gluconacetobacter xylinus/metabolism
Mice
Microscopy, Atomic Force
Microscopy, Electron, Scanning
Muramidase/metabolism
Plasmids/genetics
Reverse Transcriptase Polymerase Chain Reaction
Spectroscopy, Fourier Transform Infrared
Tandem Mass Spectrometry
X-Ray Diffraction
Chemicals
Biopolymers
DNA Primers
Chitin
Cellulose
Muramidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Yadav Vikas
Biomedical Engineering Department, Tufts University, Medford, Massachusetts 02155, USA.
Paniliatis Bruce J
Shi Hai
Lee Kyongbum
Cebe Peggy
Kaplan David L
References (30)
30 references, click to expand
-
AFM imaging of bacteria in liquid media immobilized on gelatin coated mica surfaces.
Ultramicroscopy. 2003 Oct-Nov;97(1-4):209-16
PMID: 12801673
-
Cellulose biosynthesis and function in bacteria.
Microbiol Rev. 1991 Mar;55(1):35-58
PMID: 2030672
-
Identification of a novel UDP-Glc:GlcNAc beta1-->4-glucosyltransferase in Lymnaea stagnalis that may be involved in the synthesis of complex-type oligosaccharide chains.
Glycobiology. 2000 Mar;10(3):263-71
PMID: 10704525
-
Communications to the editor: The structure of regenerated cellulose.
Macromolecules. 1975 Jul-Aug;8(4):563-4
PMID: 1177500
-
Engineering prokaryotic gene circuits.
FEMS Microbiol Rev. 2009 Jan;33(1):27-37
PMID: 19016883
-
Assay for hexosamine pathway intermediates (uridine diphosphate-N-acetyl amino sugars) in small samples of human muscle tissue.
Clin Chem. 2001 May;47(5):944-6
PMID: 11325903
-
Microbial cellulose--the natural power to heal wounds.
Biomaterials. 2006 Jan;27(2):145-51
PMID: 16099034
-
Biogenesis of bacterial cellulose.
Crit Rev Microbiol. 1991;17(6):435-47
PMID: 2039586
-
Mucosal drug delivery using cellulose derivatives as a functional polymer.
J Control Release. 1999 Nov 1;62(1-2):101-7
PMID: 10518641
-
Direct incorporation of glucosamine and N-acetylglucosamine into exopolymers by Gluconacetobacter xylinus (=Acetobacter xylinum) ATCC 10245: production of chitosan-cellulose and chitin-cellulose exopolymers.
Appl Environ Microbiol. 2001 Sep;67(9):3970-5
PMID: 11525993
-
Can biofuels finally take center stage?
Nat Biotechnol. 2006 Jul;24(7):777-84
PMID: 16841058
-
Bacterial cellulose as a potential scaffold for tissue engineering of cartilage.
Biomaterials. 2005 Feb;26(4):419-31
PMID: 15275816
-
Metabolic Engineering of a Pentose Metabolism Pathway in Ethanologenic Zymomonas mobilis.
Science. 1995 Jan 13;267(5195):240-3
PMID: 17791346
-
Biosynthesis of a novel polysaccharide by Acetobacter xylinum.
Int J Biol Macromol. 1994 Dec;16(6):297-300
PMID: 7727342
-
Synthesis of cellulose by Acetobacter xylinum. II. Preparation of freeze-dried cells capable of polymerizing glucose to cellulose.
Biochem J. 1954 Oct;58(2):345-52
PMID: 13208601
-
Genomics of cellulosic biofuels.
Nature. 2008 Aug 14;454(7206):841-5
PMID: 18704079
-
The future prospects of microbial cellulose in biomedical applications.
Biomacromolecules. 2007 Jan;8(1):1-12
PMID: 17206781
-
Enhancement of cellulose pellicle production by constitutively expressing vitreoscilla hemoglobin in Acetobacter xylinum.
Biotechnol Prog. 2006 Nov-Dec;22(6):1598-603
PMID: 17137307
-
Mechanism of lysozyme action.
Science. 1969 Aug 1;165(3892):454-65
PMID: 4893486
-
Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield.
Proc Natl Acad Sci U S A. 2008 Sep 16;105(37):13769-74
PMID: 18779592
-
Plant genetic engineering for biofuel production: towards affordable cellulosic ethanol.
Nat Rev Genet. 2008 Jun;9(6):433-43
PMID: 18487988
-
Enhancement of cellulose production by expression of sucrose synthase in Acetobacter xylinum.
Proc Natl Acad Sci U S A. 1999 Jan 5;96(1):14-8
PMID: 9874763
-
Genetic organization of the cellulose synthase operon in Acetobacter xylinum.
Proc Natl Acad Sci U S A. 1990 Oct;87(20):8130-4
PMID: 2146681
-
Ralstonia eutropha strain H16 as model organism for PHA metabolism and for biotechnological production of technically interesting biopolymers.
J Mol Microbiol Biotechnol. 2009;16(1-2):91-108
PMID: 18957865
-
Genetic engineering of polysaccharide structure: production of variants of xanthan gum in Xanthomonas campestris.
Biotechnol Prog. 1990 May-Jun;6(3):182-7
PMID: 1366611
-
In vivo biocompatibility of bacterial cellulose.
J Biomed Mater Res A. 2006 Feb;76(2):431-8
PMID: 16278860
-
Susceptibilities of bacterial cellulose containing N-acetylglucosamine residues for cellulolytic and chitinolytic enzymes.
Int J Biol Macromol. 1992 Dec;14(6):343-7
PMID: 1476990
-
Analysis of the lysozme-catalyzed hydrolysis and transglycosylation of N-acetyl-D-glucosamine oligomers by high-pressure liquid chromatography.
Anal Biochem. 1977 Feb;77(2):513-22
PMID: 842833
-
Bacteria engineered for fuel ethanol production: current status.
Appl Microbiol Biotechnol. 2003 Dec;63(3):258-66
PMID: 13680206
-
Wound healing--aiming for perfect skin regeneration.
Science. 1997 Apr 4;276(5309):75-81
PMID: 9082989