Inactivation of aldehyde dehydrogenase: A key factor for engineering 1,3-propanediol production by Klebsiella pneumoniae

Yanping Zhang, Yin Li, Chenyu Du, Ming Liu, Zhu'an Cao

Research output: Contribution to journalArticle

95 Citations (Scopus)

Abstract

Production of 1,3-propanediol (1,3-PD) from glycerol by Klebsiella pneumoniae is restrained by ethanol formation. The first step in the formation of ethanol from acetyl-CoA is catalyzed by aldehyde dehydrogenase (ALDH), an enzyme that competes with 1,3-PD oxidoreductase for the cofactor NADH. This study aimed to improve the production of 1,3-PD by engineering the ethanol formation pathway. An inactivation mutation of the aldA gene encoding ALDH in K. pneumoniae YMU2 was generated by insertion of a tetracycline resistance marker. Inactivation of ALDH resulted in a nearly abolished ethanol formation but a significantly improved 1,3-PD production. Metabolic flux analysis revealed that a pronounced redistribution of intracellular metabolic flux occurred. The final titer, the productivity of 1,3-PD and the yield of 1,3-PD relative to glycerol of the mutant strain reached 927.6 mmol L-1, 14.05 mmol L-1 h-1 and 0.699 mol mol-1, respectively, which were much higher than those of the parent strain. In addition, the specific 1,3-PD-producing capability (1,3-PD produced per gram of cells) of the mutant strain was 2-fold that of the parent strain due to a lower growth yield of the mutant. By increasing NADH availability, this study demonstrates an important metabolic engineering approach to improve the efficiency of oxidoreduction-coupled bioprocesses.

LanguageEnglish
Pages578-586
Number of pages9
JournalMetabolic Engineering
Volume8
Issue number6
DOIs
Publication statusPublished - 1 Nov 2006
Externally publishedYes

Fingerprint

Aldehyde Dehydrogenase
Klebsiella pneumoniae
Aldehydes
Ethanol
Glycerol
Fluxes
Metabolic engineering
Gene encoding
NAD
Metabolic Flux Analysis
Enzymes
Metabolic Engineering
Productivity
Tetracycline Resistance
Availability
Acetyl Coenzyme A
1,3-propanediol
Oxidoreductases
Tetracycline
Mutation

Cite this

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title = "Inactivation of aldehyde dehydrogenase: A key factor for engineering 1,3-propanediol production by Klebsiella pneumoniae",
abstract = "Production of 1,3-propanediol (1,3-PD) from glycerol by Klebsiella pneumoniae is restrained by ethanol formation. The first step in the formation of ethanol from acetyl-CoA is catalyzed by aldehyde dehydrogenase (ALDH), an enzyme that competes with 1,3-PD oxidoreductase for the cofactor NADH. This study aimed to improve the production of 1,3-PD by engineering the ethanol formation pathway. An inactivation mutation of the aldA gene encoding ALDH in K. pneumoniae YMU2 was generated by insertion of a tetracycline resistance marker. Inactivation of ALDH resulted in a nearly abolished ethanol formation but a significantly improved 1,3-PD production. Metabolic flux analysis revealed that a pronounced redistribution of intracellular metabolic flux occurred. The final titer, the productivity of 1,3-PD and the yield of 1,3-PD relative to glycerol of the mutant strain reached 927.6 mmol L-1, 14.05 mmol L-1 h-1 and 0.699 mol mol-1, respectively, which were much higher than those of the parent strain. In addition, the specific 1,3-PD-producing capability (1,3-PD produced per gram of cells) of the mutant strain was 2-fold that of the parent strain due to a lower growth yield of the mutant. By increasing NADH availability, this study demonstrates an important metabolic engineering approach to improve the efficiency of oxidoreduction-coupled bioprocesses.",
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Inactivation of aldehyde dehydrogenase : A key factor for engineering 1,3-propanediol production by Klebsiella pneumoniae. / Zhang, Yanping; Li, Yin; Du, Chenyu; Liu, Ming; Cao, Zhu'an.

In: Metabolic Engineering, Vol. 8, No. 6, 01.11.2006, p. 578-586.

Research output: Contribution to journalArticle

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T1 - Inactivation of aldehyde dehydrogenase

T2 - Metabolic Engineering

AU - Zhang, Yanping

AU - Li, Yin

AU - Du, Chenyu

AU - Liu, Ming

AU - Cao, Zhu'an

PY - 2006/11/1

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AB - Production of 1,3-propanediol (1,3-PD) from glycerol by Klebsiella pneumoniae is restrained by ethanol formation. The first step in the formation of ethanol from acetyl-CoA is catalyzed by aldehyde dehydrogenase (ALDH), an enzyme that competes with 1,3-PD oxidoreductase for the cofactor NADH. This study aimed to improve the production of 1,3-PD by engineering the ethanol formation pathway. An inactivation mutation of the aldA gene encoding ALDH in K. pneumoniae YMU2 was generated by insertion of a tetracycline resistance marker. Inactivation of ALDH resulted in a nearly abolished ethanol formation but a significantly improved 1,3-PD production. Metabolic flux analysis revealed that a pronounced redistribution of intracellular metabolic flux occurred. The final titer, the productivity of 1,3-PD and the yield of 1,3-PD relative to glycerol of the mutant strain reached 927.6 mmol L-1, 14.05 mmol L-1 h-1 and 0.699 mol mol-1, respectively, which were much higher than those of the parent strain. In addition, the specific 1,3-PD-producing capability (1,3-PD produced per gram of cells) of the mutant strain was 2-fold that of the parent strain due to a lower growth yield of the mutant. By increasing NADH availability, this study demonstrates an important metabolic engineering approach to improve the efficiency of oxidoreduction-coupled bioprocesses.

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