Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain
Edited by Jef D. Boeke, New York University School of Medicine, New York, NY, and approved April 29, 2016 (received for review November 20, 2015)
Significance
Bacterial cellulose is a remarkable material that is malleable, biocompatible, and over 10-times stronger than plant-based cellulose. It is currently used to create materials for tissue engineering, medicine, defense, electronics, acoustics, and fabrics. We describe here a bacterial strain that is readily amenable to genetic engineering and produces high quantities of bacterial cellulose in low-cost media. To reprogram this organism for biotechnology applications, we created a set of genetic tools that enables biosynthesis of patterned cellulose, functionalization of the cellulose surface with proteins, and tunable control over cellulose production. This greatly expands our ability to control and engineer new cellulose-based biomaterials, offering numerous applications for basic research, materials science, and biotechnology.
Abstract
Bacterial cellulose is a strong and ultrapure form of cellulose produced naturally by several species of the Acetobacteraceae. Its high strength, purity, and biocompatibility make it of great interest to materials science; however, precise control of its biosynthesis has remained a challenge for biotechnology. Here we isolate a strain of Komagataeibacter rhaeticus (K. rhaeticus iGEM) that can produce cellulose at high yields, grow in low-nitrogen conditions, and is highly resistant to toxic chemicals. We achieved external control over its bacterial cellulose production through development of a modular genetic toolkit that enables rational reprogramming of the cell. To further its use as an organism for biotechnology, we sequenced its genome and demonstrate genetic circuits that enable functionalization and patterning of heterologous gene expression within the cellulose matrix. This work lays the foundations for using genetic engineering to produce cellulose-based materials, with numerous applications in basic science, materials engineering, and biotechnology.
Data Availability
Data deposition: The sequence reported in this paper has been deposited in the European Nucleotide Archive, www.ebi.ac.uk/ena (accession no. PRJEB10933).
Acknowledgments
We thank Dr. Cheng-Kang Lee from the National Taiwan University of Science and Technology and Dr. Jyh-Ming Wu from Chinese Culture University for sharing plasmid pBla-Vhb-122; Ms. Victoria Geaney from the Royal College of Art, London, for help with functionalized garments; Dr. Koon-Yang Lee (Imperial College London) for helpful advice and feedback on the manuscript; Dr. Takayuki Homma (Imperial College London) for help with genome library preparation; Dr. Laurence Game (Imperial College London) for help and advice with genome sequencing; Mr. Geraint Barton (Imperial College London) for help and advice with genome assembly and analysis; Dr. Francesca Ceroni (Imperial College London) for providing ATc-inducible constructs and advice; Ms. Catherine Ainsworth, Mr. Nicolas Kral, Dr. Geoff Baldwin, and Dr. Guy-Bart Stan of Imperial College London for instruction and helpful discussions throughout the project; Dr. Mahmoud Ardakani (Imperial College London) for taking SEM images; and Dr. Carlos Bricio Garberi (Imperial College London) for taking pellicle photographs.
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Freely available online through the PNAS open access option.
Data Availability
Data deposition: The sequence reported in this paper has been deposited in the European Nucleotide Archive, www.ebi.ac.uk/ena (accession no. PRJEB10933).
Submission history
Published online: May 31, 2016
Published in issue: June 14, 2016
Keywords
Acknowledgments
We thank Dr. Cheng-Kang Lee from the National Taiwan University of Science and Technology and Dr. Jyh-Ming Wu from Chinese Culture University for sharing plasmid pBla-Vhb-122; Ms. Victoria Geaney from the Royal College of Art, London, for help with functionalized garments; Dr. Koon-Yang Lee (Imperial College London) for helpful advice and feedback on the manuscript; Dr. Takayuki Homma (Imperial College London) for help with genome library preparation; Dr. Laurence Game (Imperial College London) for help and advice with genome sequencing; Mr. Geraint Barton (Imperial College London) for help and advice with genome assembly and analysis; Dr. Francesca Ceroni (Imperial College London) for providing ATc-inducible constructs and advice; Ms. Catherine Ainsworth, Mr. Nicolas Kral, Dr. Geoff Baldwin, and Dr. Guy-Bart Stan of Imperial College London for instruction and helpful discussions throughout the project; Dr. Mahmoud Ardakani (Imperial College London) for taking SEM images; and Dr. Carlos Bricio Garberi (Imperial College London) for taking pellicle photographs.
Notes
This article is a PNAS Direct Submission.
Authors
Competing Interests
Conflict of interest statement: H.H. and G.S. are researching the industrial uses of cellulose functionalized with cellulose binding domain fusion proteins as part of CustoMem Ltd.
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Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain, Proc. Natl. Acad. Sci. U.S.A.
113 (24) E3431-E3440,
https://doi.org/10.1073/pnas.1522985113
(2016).
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