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 in | PloS one Vol. 9; no. 6; p. e98772 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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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. |
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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 |
AuthorAffiliation_xml | – name: 2 DeFENS, Department of Food Environmental and Nutritional Sciences, University of Milan, Milan, Italy – name: 3 Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin, People’s Republic of China – name: 1 Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People’s Republic of China – name: University of Groningen, Netherlands |
Author_xml | – sequence: 1 givenname: Cheng surname: Zhong fullname: Zhong, Cheng organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China – sequence: 2 givenname: Fei surname: Li fullname: Li, Fei organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China; DeFENS, Department of Food Environmental and Nutritional Sciences, University of Milan, Milan, Italy – sequence: 3 givenname: Miao surname: Liu fullname: Liu, Miao organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China – sequence: 4 givenname: Xiao-Ning surname: Yang fullname: Yang, Xiao-Ning organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China – sequence: 5 givenname: Hui-Xia surname: Zhu fullname: Zhu, Hui-Xia organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China – sequence: 6 givenname: Yuan-Yuan surname: Jia fullname: Jia, Yuan-Yuan organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China; Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin, People's Republic of China – sequence: 7 givenname: Shi-Ru surname: Jia fullname: Jia, Shi-Ru organization: Key Laboratory of Industrial Fermentation Microbiology, (Ministry of Education), Tianjin University of Science & Technology, Tianjin, People's Republic of China – sequence: 8 givenname: Luciano surname: Piergiovanni fullname: Piergiovanni, Luciano organization: DeFENS, Department of Food Environmental and Nutritional Sciences, University of Milan, Milan, Italy |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/24901455$$D View this record in MEDLINE/PubMed |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 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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Title | Revealing differences in metabolic flux distributions between a mutant strain and its parent strain Gluconacetobacter xylinus CGMCC 2955 |
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