Evolution of D-lactate dehydrogenase activity from glycerol dehydrogenase and its utility for D-lactate production from lignocellulose

Wang, Qingzhao; Ingram, L. O.; Shanmugam, K. T.

doi:10.1073/pnas.1111085108

Cited by 69 publications

(74 citation statements)

References 37 publications

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“…This finding suggests that gldA is a more ancient gene and may serve as a backup that can turn into a more efficient catalyst under selective pressure, pro- viding a survival benefit to the organism. A previous study provides considerable support for our hypotheses (55). GDH (encoded by gldA) from Bacillus coagulans acquired D-lactate dehydrogenase activity through mutations during adaptive evolution.…”

Section: Discussionsupporting

confidence: 57%

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Glycerol Dehydrogenase Plays a Dual Role in Glycerol Metabolism and 2,3-Butanediol Formation in Klebsiella pneumoniae

Wang¹,

Tao²,

2014

Journal of Biological Chemistry

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Background: Glycerol dehydrogenase catalyzes the initial step of glycerol oxidation. Results: DhaD, a glycerol dehydrogenase from Klebsiella pneumoniae, plays a dual role in glycerol metabolism and 2,3-butanediol formation. Conclusion: This dual role related to the enzyme promiscuity switches according to physiological requirements, suggesting a mechanism involved in evolution of the enzyme. Significance: This work highlights the sophistication of enzyme evolution.

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Section: Discussionsupporting

confidence: 57%

“…GDH (encoded by gldA) from Bacillus coagulans acquired D-lactate dehydrogenase activity through mutations during adaptive evolution. The expression level of gldA was also increased as a result of an upstream transposon insertion, facilitating production of D-lactic acid and restoring anaerobic growth (55).…”

Section: Discussionmentioning

confidence: 97%

Glycerol Dehydrogenase Plays a Dual Role in Glycerol Metabolism and 2,3-Butanediol Formation in Klebsiella pneumoniae

Wang¹,

Tao²,

2014

Journal of Biological Chemistry

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“…More recently, D-lactic acid formation in Synechocystis has also been accomplished (9). In the latter study, D-lactic acid formation was achieved via the involvement of a mutated glycerol dehydrogenase, GlyDH*, from Bacillus coagulans (14). Here, we have chosen to use the D-LDH of Leuconostoc mesenteroides (here L. mesenteroides) (15), because of its superior kinetics compared to other characterized D-LDH enzymes (16).…”

mentioning

confidence: 99%

Chirality Matters: Synthesis and Consumption of the d -Enantiomer of Lactic Acid by Synechocystis sp. Strain PCC6803

Angermayr

Woude²,

Correddu

et al. 2016

Appl Environ Microbiol

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growth stages. We show that Synechocystis is able to use D-lactic acid, but not L-lactic acid, as a carbon source for growth. Deletion of the organism's putative D-lactate dehydrogenase (encoded by slr1556), however, does not eliminate this ability with respect to D-lactic acid consumption. In contrast, D-lactic acid consumption does depend on the presence of glycolate dehydrogenase GlcD1 (encoded by sll0404). Accordingly, this report highlights the need to match a product of interest of a cyanobacterial cell factory with the metabolic network present in the host used for its synthesis and emphasizes the need to understand the physiology of the production host in detail.T o date, lactic acid has been produced with almost 100% conversion efficiency by several chemotrophic fermentative bacterial and yeast cell factories growing on various sugars (1, 2). Consequently, a large amount of effort is currently exerted to produce lactic acid with lactic acid bacteria (3), in combination with the use of a cheap substrate(s) such as lignocellulosic feedstock, though that feedstock requires an energy-intensive pretreatment of the biomass (1, 4). Lactic acid is used in food preservation, in the chemical and pharmaceutical industries, and as a building block for construction of polymers, the latter for use as an alternative to petroleum-derived plastics. It is compelling that the biodegradability and heat stability of this (bio)plastic depend on the blend of the two optically active, chiral isoforms of lactic acid (5).Employing a photosynthetic microorganism as the production host has the advantage of enabling the direct conversion of CO 2 into (poly)lactic acid (6-9). Such production, which is dependent on cyanobacterial cell factories, allows compound formation without the need to generate complex (plant) biomass first, only to break it down again later for its utilization by a chemotrophic fermentative microorganism (10).For living organisms, chirality plays an essential role. For example, amino acids are incorporated as L-enantiomers into proteins. Likewise, L-lactic acid seems to be the dominant enantiomeric form of this weak acid in nature. Thus, suitable and effective production hosts for D-lactic acid are more challenging to find and construct. Nonetheless, biosynthetic routes for both enantiomers, and for the corresponding products, exist in various (micro)organisms, facilitated by enantiomer-specific lactate dehydrogenases (LDH) (11), whereas chemical synthesis routinely results in a racemic mixture (12). Earlier, we constructed several L-lactic acid-producing Synechocystis sp. strain PCC6803 (here Synechocystis) variants (7, 13). In the framework of those experiments, we also tested the D-LDH of Escherichia coli (7), but we were not successful in producing D-lactic acid in the engineered Synechocystis strains at that time. However, in another cyanobacterium, Synechococcus elongatus PCC7942, synthesis of the latter enantiomer has been achieved through the expression of the same E. coli enzyme, although its extracell...

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“…They potentially can be used as platform organisms if they are genetically accessible. A number of facultatively anaerobic thermophilic hosts have been studied for green chemical and fuel production (reviewed in references 7 and 8), such as Bacillus coagulans for lactic acid (9,10), Bacillus licheniformis for 2,3-butanediol (11,12), and Geobacillus thermoglucosidans for ethanol production (13). Examples of strictly anaerobic thermophiles that have been studied for biofuel production are Clostridium thermocellum (14,15), several Caldicellulosiruptor spp.…”

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confidence: 99%

Isolation and Screening of Thermophilic Bacilli from Compost for Electrotransformation and Fermentation: Characterization of Bacillus smithii ET 138 as a New Biocatalyst

Bosma

Weijer

Daas

et al. 2015

Appl Environ Microbiol

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b Thermophilic bacteria are regarded as attractive production organisms for cost-efficient conversion of renewable resources to green chemicals, but their genetic accessibility is a major bottleneck in developing them into versatile platform organisms. In this study, we aimed to isolate thermophilic, facultatively anaerobic bacilli that are genetically accessible and have potential as platform organisms. From compost, we isolated 267 strains that produced acids from C 5 and C 6 sugars at temperatures of 55°C or 65°C. Subsequently, 44 strains that showed the highest production of acids were screened for genetic accessibility by electroporation. Two Geobacillus thermodenitrificans isolates and one Bacillus smithii isolate were found to be transformable with plasmid pNW33n. Of these, B. smithii ET 138 was the best-performing strain in laboratory-scale fermentations and was capable of producing organic acids from glucose as well as from xylose. It is an acidotolerant strain able to produce organic acids until a lower limit of approximately pH 4.5. As genetic accessibility of B. smithii had not been described previously, six other B. smithii strains from the DSMZ culture collection were tested for electroporation efficiencies, and we found the type strain DSM 4216 T and strain DSM 460 to be transformable. The transformation protocol for B. smithii isolate ET 138 was optimized to obtain approximately 5 ؋ 10 3 colonies per g plasmid pNW33n. Genetic accessibility combined with robust acid production capacities on C 5 and C 6 sugars at a relatively broad pH range make B. smithii ET 138 an attractive biocatalyst for the production of lactic acid and potentially other green chemicals. Green chemicals are sustainable bio-based alternatives for chemicals based on fossil resources. Biomass is considered an attractive renewable resource for the production of such green chemicals. Major challenges for using biomass are to develop costeffective production processes and to convert substrates that go beyond first-generation pure sugars. From this perspective, the use of microbial fermentation processes that convert lignocellulosic sugars is gaining considerable attention. For particular products natural producers can be used, but often Escherichia coli and Saccharomyces cerevisiae are used as platform organisms because of their well-known physiology and ease of engineering for the production of many different products (1, 2).In general, most organisms used for the production of green chemicals and fuels are mesophiles, and current processes still are based mainly on first-generation feedstocks, i.e., sucrose from sugar beet or sugarcane or glucose derived from starches from corn or tapioca. Although the engineering of platform organisms broadens the spectrum of possible substrates (3), the efficiency of the process could be increased by the use of organisms naturally capable of degrading lignocellulose-derived sugars (4). To further increase the efficiency and reduce the costs of the microbial production of green chemicals, modera...

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Evolution of D-lactate dehydrogenase activity from glycerol dehydrogenase and its utility for D-lactate production from lignocellulose

Cited by 69 publications

References 37 publications

Glycerol Dehydrogenase Plays a Dual Role in Glycerol Metabolism and 2,3-Butanediol Formation in Klebsiella pneumoniae

Glycerol Dehydrogenase Plays a Dual Role in Glycerol Metabolism and 2,3-Butanediol Formation in Klebsiella pneumoniae

Chirality Matters: Synthesis and Consumption of the d -Enantiomer of Lactic Acid by Synechocystis sp. Strain PCC6803

Isolation and Screening of Thermophilic Bacilli from Compost for Electrotransformation and Fermentation: Characterization of Bacillus smithii ET 138 as a New Biocatalyst

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