Revealing differences in metabolic flux distributions between a mutant strain and its parent strain Gluconacetobacter xylinus CGMCC 2955

A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain (G. xylinus CGMCC 2955)...

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Published inPloS one Vol. 9; no. 6; p. e98772
Main Authors Zhong, Cheng, Li, Fei, Liu, Miao, Yang, Xiao-Ning, Zhu, Hui-Xia, Jia, Yuan-Yuan, Jia, Shi-Ru, Piergiovanni, Luciano
Format Journal Article
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Published United States Public Library of Science 05.06.2014
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Abstract A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain (G. xylinus CGMCC 2955) using DEC (diethyl sulfate) and LiCl. The highest bacterial cellulose production for this mutant was obtained at about 11.75 g/L, which was an increase of 62% compared with that by the parent strain. In contrast, gluconic acid (the main byproduct) concentration was only 5.71 g/L for mutant strain, which was 55.7% lower than that of parent strain. Metabolic flux analysis indicated that 40.1% of the carbon source was transformed to bacterial cellulose in mutant strain, compared with 24.2% for parent strain. Only 32.7% and 4.0% of the carbon source were converted into gluconic acid and acetic acid in mutant strain, compared with 58.5% and 9.5% of that in parent strain. In addition, a higher flux of tricarboxylic acid (TCA) cycle was obtained in mutant strain (57.0%) compared with parent strain (17.0%). It was also indicated from the flux analysis that more ATP was produced in mutant strain from pentose phosphate pathway (PPP) and TCA cycle. The enzymatic activity of succinate dehydrogenase (SDH), which is one of the key enzymes in TCA cycle, was 1.65-fold higher in mutant strain than that in parent strain at the end of culture. It was further validated by the measurement of ATPase that 3.53-6.41 fold higher enzymatic activity was obtained from mutant strain compared with parent strain.
AbstractList A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain (G. xylinus CGMCC 2955) using DEC (diethyl sulfate) and LiCl. The highest bacterial cellulose production for this mutant was obtained at about 11.75 g/L, which was an increase of 62% compared with that by the parent strain. In contrast, gluconic acid (the main byproduct) concentration was only 5.71 g/L for mutant strain, which was 55.7% lower than that of parent strain. Metabolic flux analysis indicated that 40.1% of the carbon source was transformed to bacterial cellulose in mutant strain, compared with 24.2% for parent strain. Only 32.7% and 4.0% of the carbon source were converted into gluconic acid and acetic acid in mutant strain, compared with 58.5% and 9.5% of that in parent strain. In addition, a higher flux of tricarboxylic acid (TCA) cycle was obtained in mutant strain (57.0%) compared with parent strain (17.0%). It was also indicated from the flux analysis that more ATP was produced in mutant strain from pentose phosphate pathway (PPP) and TCA cycle. The enzymatic activity of succinate dehydrogenase (SDH), which is one of the key enzymes in TCA cycle, was 1.65-fold higher in mutant strain than that in parent strain at the end of culture. It was further validated by the measurement of ATPase that 3.53-6.41 fold higher enzymatic activity was obtained from mutant strain compared with parent strain.
A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable by-products. A mutant strain, Gluconacetobacter xylinus AX2-16, was obtained by combined chemical mutation of the parent strain ( G. xylinus CGMCC 2955) using DEC (diethyl sulfate) and LiCl. The highest bacterial cellulose production for this mutant was obtained at about 11.75 g/L, which was an increase of 62% compared with that by the parent strain. In contrast, gluconic acid (the main byproduct) concentration was only 5.71 g/L for mutant strain, which was 55.7% lower than that of parent strain. Metabolic flux analysis indicated that 40.1% of the carbon source was transformed to bacterial cellulose in mutant strain, compared with 24.2% for parent strain. Only 32.7% and 4.0% of the carbon source were converted into gluconic acid and acetic acid in mutant strain, compared with 58.5% and 9.5% of that in parent strain. In addition, a higher flux of tricarboxylic acid (TCA) cycle was obtained in mutant strain (57.0%) compared with parent strain (17.0%). It was also indicated from the flux analysis that more ATP was produced in mutant strain from pentose phosphate pathway (PPP) and TCA cycle. The enzymatic activity of succinate dehydrogenase (SDH), which is one of the key enzymes in TCA cycle, was 1.65-fold higher in mutant strain than that in parent strain at the end of culture. It was further validated by the measurement of ATPase that 3.53–6.41 fold higher enzymatic activity was obtained from mutant strain compared with parent strain.
Audience Academic
Author Jia, Shi-Ru
Liu, Miao
Yang, Xiao-Ning
Jia, Yuan-Yuan
Zhong, Cheng
Li, Fei
Piergiovanni, Luciano
Zhu, Hui-Xia
AuthorAffiliation 1 Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People’s Republic of China
University of Groningen, Netherlands
2 DeFENS, Department of Food Environmental and Nutritional Sciences, University of Milan, Milan, Italy
3 Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin, People’s Republic of China
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/24901455$$D View this record in MEDLINE/PubMed
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2014 Zhong et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
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– notice: 2014 Zhong et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
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Conceived and designed the experiments: CZ FL ML SRJ. Performed the experiments: CZ FL ML XNY HXZ. Analyzed the data: CZ FL ML HXZ YYJ SRJ. Contributed reagents/materials/analysis tools: FL YYJ SRJ. Wrote the paper: CZ FL ML SRJ LP.
Competing Interests: The authors have declared that no competing interests exist.
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Snippet A better understanding of metabolic fluxes is important for manipulating microbial metabolism toward desired end products, or away from undesirable...
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SubjectTerms Acetic acid
Acids
Adenosine triphosphatase
ATPases
Bacteria
Biology and Life Sciences
Biosynthesis
Byproducts
Carbon
Carbon sources
Cell culture
Cellulose
Cellulose - biosynthesis
Citric Acid Cycle
Comparative analysis
Dehydrogenases
E coli
Education
Energy Metabolism
Enzymatic activity
Enzymes
Escherichia coli
Fermentation
Fluctuations
Fluxes
Gluconacetobacter xylinus - genetics
Gluconacetobacter xylinus - metabolism
Gluconic acid
Gluconobacter oxydans
Glucose
Glycerol
Laboratories
Lithium chloride
Metabolic flux
Metabolic Networks and Pathways
Metabolism
Metabolites
Microbiology
Microorganisms
Monosaccharides
Mutation
Organic acids
Pentose
Pentose phosphate
Pentose phosphate pathway
Phosphates
Productivity
Science
Succinate dehydrogenase
Sulfate
Sulfates
Tricarboxylic acid cycle
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Title Revealing differences in metabolic flux distributions between a mutant strain and its parent strain Gluconacetobacter xylinus CGMCC 2955
URI https://www.ncbi.nlm.nih.gov/pubmed/24901455
https://www.proquest.com/docview/1532986780
https://search.proquest.com/docview/1534101121
https://pubmed.ncbi.nlm.nih.gov/PMC4047042
https://doaj.org/article/177242057f6746e18d5b807aa01ebd75
http://dx.doi.org/10.1371/journal.pone.0098772
Volume 9
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