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Title: Mutation in galP improved fermentation of mixed sugars to succinate using engineered Escherichia coli AS1600a and AM1 mineral salts medium

Abstract

Escherichia coli KJ122 was engineered to produce succinate from glucose using the wild type GalP for glucose uptake instead of the native phosphotransferase system (ptsI mutation). This strain now ferments 10% xylose poorly. Mutants were selected by serial transfers in AM1 mineral salts medium with 10% xylose. Clones from this population all exhibited a similar improvement, co-fermentation of an equal mixture of xylose and glucose. One of these, AS1600a, produced 84.26 ± 1.37 g/L succinate, equivalent to that produced by the parent (KJ122) from 10% glucose (85.46 ± 1.78 g/L). AS1600a was sequenced and found to contain a mutation in galactose permease (GalP, G236D). This mutation was shown to be responsible for the improvement in fermentation using KJΔgalP as the host and expression vectors with native galP and with mutant galP*. Strain AS1600a and KJΔgalP(pLOI5746; galP*) also co-fermented a mixture of glucose, xylose, arabinose, and galactose in sugarcane bagasse hydrolysate using mineral salts medium.

Authors:
 [1];  [1];  [2];  [2];  [2];  [2];  [2]
  1. Suranaree Univ. of Technology, Nakhon Ratchasima (Thailand)
  2. Univ. of Florida, Gainesville, FL (United States)
Publication Date:
Research Org.:
Univ. of Florida, Gainesville, FL (United States)
Sponsoring Org.:
USDOE Office of International Affairs (IA); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office; U.S. Department of Agriculture; Thailand Research Fund (TRF)
OSTI Identifier:
1771326
Alternate Identifier(s):
OSTI ID: 1250363; OSTI ID: 1661478
Grant/Contract Number:  
PI0000031; PHD/0135/2553
Resource Type:
Accepted Manuscript
Journal Name:
Bioresource Technology
Additional Journal Information:
Journal Volume: 193; Journal ID: ISSN 0960-8524
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Succinate; Furfural; Sugarcane bagasse; Xylose; Arabinose; succinate; furfural; sugarcane bagasse; xylose; arabinose

Citation Formats

Sawisit, Apichai, Jantama, Kaemwich, Zheng, Huabao, Yomano, Lorraine P., York, Sean W., Shanmugam, Keelnatham T., and Ingram, Lonnie O. Mutation in galP improved fermentation of mixed sugars to succinate using engineered Escherichia coli AS1600a and AM1 mineral salts medium. United States: N. p., 2015. Web. doi:10.1016/j.biortech.2015.06.108.
Sawisit, Apichai, Jantama, Kaemwich, Zheng, Huabao, Yomano, Lorraine P., York, Sean W., Shanmugam, Keelnatham T., & Ingram, Lonnie O. Mutation in galP improved fermentation of mixed sugars to succinate using engineered Escherichia coli AS1600a and AM1 mineral salts medium. United States. https://doi.org/10.1016/j.biortech.2015.06.108
Sawisit, Apichai, Jantama, Kaemwich, Zheng, Huabao, Yomano, Lorraine P., York, Sean W., Shanmugam, Keelnatham T., and Ingram, Lonnie O. Thu . "Mutation in galP improved fermentation of mixed sugars to succinate using engineered Escherichia coli AS1600a and AM1 mineral salts medium". United States. https://doi.org/10.1016/j.biortech.2015.06.108. https://www.osti.gov/servlets/purl/1771326.
@article{osti_1771326,
title = {Mutation in galP improved fermentation of mixed sugars to succinate using engineered Escherichia coli AS1600a and AM1 mineral salts medium},
author = {Sawisit, Apichai and Jantama, Kaemwich and Zheng, Huabao and Yomano, Lorraine P. and York, Sean W. and Shanmugam, Keelnatham T. and Ingram, Lonnie O.},
abstractNote = {Escherichia coli KJ122 was engineered to produce succinate from glucose using the wild type GalP for glucose uptake instead of the native phosphotransferase system (ptsI mutation). This strain now ferments 10% xylose poorly. Mutants were selected by serial transfers in AM1 mineral salts medium with 10% xylose. Clones from this population all exhibited a similar improvement, co-fermentation of an equal mixture of xylose and glucose. One of these, AS1600a, produced 84.26 ± 1.37 g/L succinate, equivalent to that produced by the parent (KJ122) from 10% glucose (85.46 ± 1.78 g/L). AS1600a was sequenced and found to contain a mutation in galactose permease (GalP, G236D). This mutation was shown to be responsible for the improvement in fermentation using KJΔgalP as the host and expression vectors with native galP and with mutant galP*. Strain AS1600a and KJΔgalP(pLOI5746; galP*) also co-fermented a mixture of glucose, xylose, arabinose, and galactose in sugarcane bagasse hydrolysate using mineral salts medium.},
doi = {10.1016/j.biortech.2015.06.108},
journal = {Bioresource Technology},
number = ,
volume = 193,
place = {United States},
year = {Thu Jun 25 00:00:00 EDT 2015},
month = {Thu Jun 25 00:00:00 EDT 2015}
}

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Cited by: 26 works
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