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Title: Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440

Abstract

Pyrolysis offers a straightforward approach for the deconstruction of plant cell wall polymers into bio-oil. Recently, there has been substantial interest in bio-oil fractionation and subsequent use of biological approaches to selectively upgrade some of the resulting fractions. A fraction of particular interest for biological upgrading consists of polysaccharide-derived substrates including sugars and sugar dehydration products such as levoglucosan and cellobiosan, which are two of the most abundant pyrolysis products of cellulose. Levoglucosan can be converted to glucose-6-phosphate through the use of a levoglucosan kinase (LGK), but to date, the mechanism for cellobiosan utilization has not been demonstrated. Here, we engineer the microbe Pseudomonas putida KT2440 to use levoglucosan as a sole carbon and energy source through LGK integration. Furthermore, we demonstrate that cellobiosan can be enzymatically converted to levoglucosan and glucose with β-glucosidase enzymes from both Glycoside Hydrolase Family 1 and Family 3. β-glucosidases are commonly used in both natural and industrial cellulase cocktails to convert cellobiose to glucose to relieve cellulase product inhibition and to facilitate microbial uptake of glucose. Using an exogenous β-glucosidase, we demonstrate that the engineered strain of P. putida can grow on levoglucosan up to 60 g/L and can also utilize cellobiosan. Overall, thismore » study elucidates the biological pathway to co-utilize levoglucosan and cellobiosan, which will be a key transformation for the biological upgrading of pyrolysis-derived substrates.« less

Authors:
 [1];  [1];  [1];  [1];  [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Energy Efficiency and Renewable Energy (EERE), Bioenergy Technologies Office (EE-3B)
OSTI Identifier:
1433726
Alternate Identifier(s):
OSTI ID: 1240242
Report Number(s):
NREL/JA-5100-65971
Journal ID: ISSN 2214-0301
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Published Article
Journal Name:
Metabolic Engineering Communications
Additional Journal Information:
Journal Volume: 3; Related Information: Metabolic Engineering Communications; Journal ID: ISSN 2214-0301
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANAYLYTICAL CHEMISTRY; Pseudomonas putida KT2440; levoglucosan kinase; B-glucosidase; cellobiosan; pyrolysis; biofuels

Citation Formats

Linger, Jeffrey G., Hobdey, Sarah E., Franden, Mary Ann, Fulk, Emily M., and Beckham, Gregg T.. Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440. United States: N. p., 2016. Web. doi:10.1016/j.meteno.2016.01.005.
Linger, Jeffrey G., Hobdey, Sarah E., Franden, Mary Ann, Fulk, Emily M., & Beckham, Gregg T.. Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440. United States. doi:10.1016/j.meteno.2016.01.005.
Linger, Jeffrey G., Hobdey, Sarah E., Franden, Mary Ann, Fulk, Emily M., and Beckham, Gregg T.. Tue . "Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440". United States. doi:10.1016/j.meteno.2016.01.005.
@article{osti_1433726,
title = {Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440},
author = {Linger, Jeffrey G. and Hobdey, Sarah E. and Franden, Mary Ann and Fulk, Emily M. and Beckham, Gregg T.},
abstractNote = {Pyrolysis offers a straightforward approach for the deconstruction of plant cell wall polymers into bio-oil. Recently, there has been substantial interest in bio-oil fractionation and subsequent use of biological approaches to selectively upgrade some of the resulting fractions. A fraction of particular interest for biological upgrading consists of polysaccharide-derived substrates including sugars and sugar dehydration products such as levoglucosan and cellobiosan, which are two of the most abundant pyrolysis products of cellulose. Levoglucosan can be converted to glucose-6-phosphate through the use of a levoglucosan kinase (LGK), but to date, the mechanism for cellobiosan utilization has not been demonstrated. Here, we engineer the microbe Pseudomonas putida KT2440 to use levoglucosan as a sole carbon and energy source through LGK integration. Furthermore, we demonstrate that cellobiosan can be enzymatically converted to levoglucosan and glucose with β-glucosidase enzymes from both Glycoside Hydrolase Family 1 and Family 3. β-glucosidases are commonly used in both natural and industrial cellulase cocktails to convert cellobiose to glucose to relieve cellulase product inhibition and to facilitate microbial uptake of glucose. Using an exogenous β-glucosidase, we demonstrate that the engineered strain of P. putida can grow on levoglucosan up to 60 g/L and can also utilize cellobiosan. Overall, this study elucidates the biological pathway to co-utilize levoglucosan and cellobiosan, which will be a key transformation for the biological upgrading of pyrolysis-derived substrates.},
doi = {10.1016/j.meteno.2016.01.005},
journal = {Metabolic Engineering Communications},
number = ,
volume = 3,
place = {United States},
year = {2016},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1016/j.meteno.2016.01.005

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