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Modular Optimization of a Hemicellulose-Utilizing Pathway in Corynebacterium glutamicum for Consolidated Bioprocessing of Hemicellulosic Biomass Article Options
Sung Sun Yim†, Jae Woong Choi†, Se Hwa Lee†, and Ki Jun Jeong*†‡ † Department of Chemical and Biomolecular Engineering, BK21 Plus Program, ‡ Institute for the BioCentury, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
Cite this: ACS Synth. Biol. 5, 4, 334-343
ACS Synth. Biol., 2016, 5 (4), pp 334–343 DOI: 10.1021/acssynbio.5b00228 Publication Date (Web): January 25, 2016 Copyright © 2016 American Chemical Society *Telephone: +82-42-350-3934. Fax: +82-42-350-3910. E-mail:
[email protected].
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Hemicellulose, which is the second most abundant polysaccharide in nature after cellulose, has
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the potential to become a major feedstock for microbial fermentation to produce various biofuels and chemicals. To utilize hemicellulose economically, it is necessary to develop a consolidated bioprocess (CBP), in which all processes from biomass degradation to the production of target products occur in a single bioreactor. Here, we report a modularly engineered Corynebacterium
Yim, Sung Sun
glutamicum strain suitable for CBP using hemicellulosic biomass (xylan) as a feedstock. The hemicellulose-utilizing pathway was divided into three distinct modules, and each module was
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separately optimized. In the module for xylose utilization, the expression level of the xylose isomerase (xylA) and xylulokinase (xylB) genes was optimized with synthetic promoters of different strengths. Then, the module for xylose transport was engineered with combinatorial
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sets of synthetic promoters and heterologous transporters to achieve the fastest cell growth rate on xylose (0.372 h–1). Next, the module for the enzymatic degradation of xylan to xylose was
Received 8 November 2015 Published online 25 January 2016 Published in print 15 April 2016
also engineered with different combinations of promoters and signal peptides to efficiently secrete both endoxylanase and xylosidase into the extracellular medium. Finally, each optimized module was integrated into a single plasmid to construct a highly efficient xylan-utilizing pathway. Subsequently, the direct production of lysine from xylan was successfully
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demonstrated with the engineered pathway. To the best of our knowledge, this is the first report
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of the development of a consolidated bioprocessing C. glutamicum strain for hemicellulosic
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biomass. Keywords: consolidated bioprocess; Corynebacterium glutamicum; hemicellulose; modular engineering; synthetic promoter; CU RREN T IS S U E
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