scholarly journals Engineering a Cyanobacterial Cell Factory for Production of Lactic Acid

2012 ◽  
Vol 78 (19) ◽  
pp. 7098-7106 ◽  
Author(s):  
S. Andreas Angermayr ◽  
Michal Paszota ◽  
Klaas J. Hellingwerf
Keyword(s):  
Lactic Acid ◽  
Lactate Dehydrogenase ◽  
Lactic Acid Production ◽  
Lactate Production ◽  
Cell Factory ◽  
Wild Type ◽  
Extracellular Medium ◽  
Content Type ◽  
Bulk Chemicals ◽  
Versatile Tool

ABSTRACTMetabolic engineering of microorganisms has become a versatile tool to facilitate production of bulk chemicals, fuels, etc. Accordingly, CO2has been exploited via cyanobacterial metabolism as a sustainable carbon source of biofuel and bioplastic precursors. Here we extended these observations by showing that integration of anldhgene fromBacillus subtilis(encoding anl-lactate dehydrogenase) into the genome ofSynechocystissp. strain PCC6803 leads tol-lactic acid production, a phenotype which is shown to be stable for prolonged batch culturing. Coexpression of a heterologous soluble transhydrogenase leads to an even higher lactate production rate and yield (lactic acid accumulating up to a several-millimolar concentration in the extracellular medium) than those for the singleldhmutant. The expression of a transhydrogenase alone, however, appears to be harmful to the cells, and a mutant carrying such a gene is rapidly outcompeted by a revertant(s) with a wild-type growth phenotype. Furthermore, our results indicate that the introduction of a lactate dehydrogenase rescues this phenotype by preventing the reversion.

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

2015 ◽  
Vol 82 (4) ◽  
pp. 1295-1304 ◽  
Author(s):  
S. Andreas Angermayr ◽  
Aniek D. van der Woude ◽  
Danilo Correddu ◽  
Ramona Kern ◽  
Martin Hagemann ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactate Dehydrogenase ◽  
Acid Production ◽  
Lactic Acid Production ◽  
Cell Factory ◽  
Growth Stages ◽  
Microbial Cell Factory ◽  
Content Type ◽  
Acid Consumption ◽  
Glycolate Dehydrogenase

ABSTRACTBoth enantiomers of lactic acid,l-lactic acid andd-lactic acid, can be produced in a sustainable way by a photosynthetic microbial cell factory and thus from CO2, sunlight, and water. Several properties of polylactic acid (a polyester of polymerized lactic acid) depend on the controlled blend of these two enantiomers. Recently, cyanobacteriumSynechocystissp. strain PCC6803 was genetically modified to allow formation of either of these two enantiomers. This report elaborates on thed-lactic acid production achieved by the introduction of ad-specific lactate dehydrogenase from the lactic acid bacteriumLeuconostoc mesenteroidesintoSynechocystis. A typical batch culture of this recombinant strain initially shows lactic acid production, followed by a phase of lactic acid consumption, until production “outcompetes” consumption at later growth stages. We show thatSynechocystisis able to used-lactic acid, but notl-lactic acid, as a carbon source for growth. Deletion of the organism's putatived-lactate dehydrogenase (encoded byslr1556), however, does not eliminate this ability with respect tod-lactic acid consumption. In contrast,d-lactic acid consumption does depend on the presence of glycolate dehydrogenase GlcD1 (encoded bysll0404). 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.

scholarly journals Major Role of NAD-Dependent Lactate Dehydrogenases in the Production of l-Lactic Acid with High Optical Purity by the Thermophile Bacillus coagulans

2014 ◽  
Vol 80 (23) ◽  
pp. 7134-7141 ◽  
Author(s):  
Limin Wang ◽  
Yumeng Cai ◽  
Lingfeng Zhu ◽  
Honglian Guo ◽  
Bo Yu
Keyword(s):  
Lactic Acid ◽  
Lactate Dehydrogenase ◽  
Acid Production ◽  
Lactic Acid Production ◽  
Optical Purity ◽  
Bacillus Coagulans ◽  
Lactobacillus Delbrueckii ◽  
Content Type ◽  
High Optical Purity ◽  
Optically Pure

ABSTRACTBacillus coagulans2-6 is an excellent producer of optically purel-lactic acid. However, little is known about the mechanism of synthesis of the highly optically purel-lactic acid produced by this strain. Three enzymes responsible for lactic acid production—NAD-dependentl-lactate dehydrogenase (l-nLDH; encoded byldhL), NAD-dependentd-lactate dehydrogenase (d-nLDH; encoded byldhD), and glycolate oxidase (GOX)—were systematically investigated in order to study the relationship between these enzymes and the optical purity of lactic acid.Lactobacillus delbrueckiisubsp.bulgaricusDSM 20081 (ad-lactic acid producer) andLactobacillus plantarumsubsp.plantarumDSM 20174 (adl-lactic acid producer) were also examined in this study as comparative strains, in addition toB. coagulans. The specific activities of key enzymes for lactic acid production in the three strains were characterizedin vivoandin vitro, and the levels of transcription of theldhL,ldhD, and GOX genes during fermentation were also analyzed. The catalytic activities ofl-nLDH andd-nLDH were different inl-,d-, anddl-lactic acid producers. Onlyl-nLDH activity was detected inB. coagulans2-6 under native conditions, and the level of transcription ofldhLinB. coagulans2-6 was much higher than that ofldhDor the GOX gene at all growth phases. However, for the twoLactobacillusstrains used in this study,ldhDtranscription levels were higher than those ofldhL. The high catalytic efficiency ofl-nLDH toward pyruvate and the high transcription ratios ofldhLtoldhDandldhLto the GOX gene provide the key explanations for the high optical purity ofl-lactic acid produced byB. coagulans2-6.

scholarly journals Synthesis and initial screening of lactate dehydrogenase inhibitor activity of 1,3-benzodioxole derivatives

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Dicky Annas ◽  
Se-Yun Cheon ◽  
Mohammad Yusuf ◽  
Sung-Jin Bae ◽  
Ki-Tae Ha ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactate Dehydrogenase ◽  
Cancer Cells ◽  
Aerobic Glycolysis ◽  
Dominant Role ◽  
Lactic Acid Production ◽  
Human Pancreatic Cancer ◽  
Drug Candidate ◽  
Acid Fermentation ◽  
High Level

AbstractCancer is one of the main causes of mortality in the world. Many cancer cells produce ATP through high-level lactic acid fermentation catalyzed by lactate dehydrogenase (LDH), which converts pyruvic acid to lactic acid. LDH plays a dominant role in the Warburg effect, wherein aerobic glycolysis is favored over oxidative phosphorylation. Due to the high lactic acid production level in cancer cells, LDH-targeting could be a potential cancer treatment strategy. A few approaches, such as drug treatment, reportedly inhibited LDH activity. In this study, we describe new 1,3-benzodioxole derivatives that might be potential small molecule candidates for LDHA inhibition. The synthesis was carried out by trans-esterification between aryl ester and alcohol groups from piperonyl alcohol. Compounds 2 and 10 exhibited a selective LDHA IC50 value of 13.63 µM and 47.2 µM, respectively. Whereas only compound 10 showed significant cytotoxicity in several lines of cancer cells, especially in human pancreatic cancer PANC-1 cells. These synthesized compounds possess 2 aromatic rings and –CF3 moiety, which expectedly contributes to LDHA inhibition. The presented products have the potential to become a promising LDHA inhibitor drug candidate.

scholarly journals Isolation and Expression of Lactate Dehydrogenase Genes from Rhizopus oryzae

2000 ◽  
Vol 66 (6) ◽  
pp. 2343-2348 ◽  
Author(s):  
Christopher D. Skory
Keyword(s):  
Lactic Acid ◽  
Nucleotide Sequence Identity ◽  
Rhizopus Oryzae ◽  
Lactic Acid Production ◽  
Lactate Production ◽  
Protein Sequencing ◽  
Sequence Comparisons ◽  
Sequence Identity ◽  
Fermentative Growth ◽  
Lactate Dehydrogenases

ABSTRACT Rhizopus oryzae is used for industrial production of lactic acid, yet little is known about the genetics of this fungus. In this study I cloned two genes, ldhA and ldhB, which code for NAD+-dependent l-lactate dehydrogenases (LDH) (EC 1.1.1.27 ), from a lactic acid-producing strain of R. oryzae. These genes are similar to each other and exhibit more than 90% nucleotide sequence identity and they contain no introns. This is the first description of ldh genes in a fungus, and sequence comparisons revealed that these genes are distinct from previously isolated prokaryotic and eukaryotic ldh genes. Protein sequencing of the LDH isolated from R. oryzae during lactic acid production confirmed that ldhA codes for a 36-kDa protein that converts pyruvate to lactate. Production of LdhA was greatest when glucose was the carbon source, followed by xylose and trehalose; all of these sugars could be fermented to lactic acid. Transcripts fromldhB were not detected when R. oryzae was grown on any of these sugars but were present when R. oryzae was grown on glycerol, ethanol, and lactate. I hypothesize thatldhB encodes a second NAD+-dependent LDH that is capable of converting l-lactate to pyruvate and is produced by cultures grown on these nonfermentable substrates. BothldhA and ldhB restored fermentative growth toEscherichia coli (ldhA pfl) mutants so that they grew anaerobically and produced lactic acid.

scholarly journals Metabolic Engineering of Lactobacillus helveticus CNRZ32 for Production of Purel-(+)-Lactic Acid

2000 ◽  
Vol 66 (9) ◽  
pp. 3835-3841 ◽  
Author(s):  
Kari Kyl�-Nikkil� ◽  
Mervi Hujanen ◽  
Matti Leisola ◽  
Airi Palva
Keyword(s):  
Lactic Acid ◽  
Structural Gene ◽  
Acid Production ◽  
Wild Type Strain ◽  
Lactic Acid Production ◽  
Gene Replacement ◽  
Lactobacillus Helveticus ◽  
Statistical Experimental Design ◽  
Wild Type ◽  
In The Wild

ABSTRACT Expression of d-(−)-lactate dehydrogenase (d-LDH) and l-(+)-LDH genes (ldhDand ldhL, respectively) and production ofd-(−)- and l-(+)-lactic acid were studied inLactobacillus helveticus CNRZ32. In order to develop a host for production of pure l-(+)-isomer of lactic acid, twoldhD-negative L. helveticus CNRZ32 strains were constructed using gene replacement. One of the strains was constructed by deleting the promoter region of the ldhD gene, and the other was constructed by replacing the structural gene ofldhD with an additional copy of the structural gene (ldhL) of l-LDH of the same species. The resulting strains were designated GRL86 and GRL89, respectively. In strain GRL89, the second copy of the ldhL structural gene was expressed under the ldhD promoter. The twod-LDH-negative strains produced onlyl-(+)-lactic acid in an amount equal to the total lactate produced by the wild type. The maximum l-LDH activity was found to be 53 and 93% higher in GRL86 and GRL89, respectively, than in the wild-type strain. Furthermore, process variables forl-(+)-lactic acid production by GRL89 were optimized using statistical experimental design and response surface methodology. The temperature and pH optima were 41�C and pH 5.9. At low pH, when the growth and lactic acid production are uncoupled, strain GRL89 produced approximately 20% more lactic acid than GRL86.

scholarly journals Lactate Dehydrogenase Is the Key Enzyme for Pneumococcal Pyruvate Metabolism and Pneumococcal Survival in Blood

2014 ◽  
Vol 82 (12) ◽  
pp. 5099-5109 ◽  
Author(s):  
Paula Gaspar ◽  
Firas A. Y. Al-Bayati ◽  
Peter W. Andrew ◽  
Ana Rute Neves ◽  
Hasan Yesilkaya
Keyword(s):  
Lactate Dehydrogenase ◽  
Lactate Production ◽  
Virulence Gene ◽  
Redox Balance ◽  
Central Metabolism ◽  
Content Type ◽  
Virulence Gene Expression ◽  
Key Enzyme

ABSTRACTStreptococcus pneumoniaeis a fermentative microorganism and causes serious diseases in humans, including otitis media, bacteremia, meningitis, and pneumonia. However, the mechanisms enabling pneumococcal survival in the host and causing disease in different tissues are incompletely understood. The available evidence indicates a strong link between the central metabolism and pneumococcal virulence. To further our knowledge on pneumococcal virulence, we investigated the role of lactate dehydrogenase (LDH), which converts pyruvate to lactate and is an essential enzyme for redox balance, in the pneumococcal central metabolism and virulence using an isogenicldhmutant. Loss of LDH led to a dramatic reduction of the growth rate, pinpointing the key role of this enzyme in fermentative metabolism. The pattern of end products was altered, and lactate production was totally blocked. The fermentation profile was confirmed byin vivonuclear magnetic resonance (NMR) measurements of glucose metabolism in nongrowing cell suspensions of theldhmutant. In this strain, a bottleneck in the fermentative steps is evident from the accumulation of pyruvate, revealing LDH as the most efficient enzyme in pyruvate conversion. An increase in ethanol production was also observed, indicating that in the absence of LDH the redox balance is maintained through alcohol dehydrogenase activity. We also found that the absence of LDH renders the pneumococci avirulent after intravenous infection and leads to a significant reduction in virulence in a model of pneumonia that develops after intranasal infection, likely due to a decrease in energy generation and virulence gene expression.

D-Lactic Acid Production from Xylose in Engineered Escherichia coli SZ470

2013 ◽  
Vol 641-642 ◽  
pp. 721-724
Author(s):  
Zhao Min Zheng ◽  
Tian Tian ◽  
Jin Hua Wang ◽  
Yong Ze Wang ◽  
Sheng De Zhou
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Acid Production ◽  
Shake Flask ◽  
Lactic Acid Production ◽  
Lactate Production ◽  
Pyruvate Decarboxylase ◽  
Genetically Engineered ◽  
Acetate Production ◽  
A Minor

WD100, knocked out adhE of Escherichia coli SZ470 and inserted ldhA into Escherichia coli WD01, was genetically engineered to utilize xylose. D-lactate production was investigated for shake flask cultures with xylose. In 64h WD100 produce 10.1g/L D-lactate in the shaking flask And it consumed 25g/L xylose during the ending of fermentation.This volumetric productivity with xylose is 0.14 g·L-1·h-1.Because of pyruvate decarboxylase (poxB) expressed in flask fermention,acetate production was up to 4.7g/L.Succinate,formate,ethanol was also produced as a minor product during fermentation.

scholarly journals Production of biopolymer precursors beta-alanine and L-lactic acid from CO2 with metabolically versatile Rhodococcus opacus DSM 43205

2020 ◽  
Author(s):  
Laura Salusjärvi ◽  
Leo Ojala ◽  
Gopal Peddinti ◽  
Michael Lienemann ◽  
Paula Jouhten ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Metabolic Engineering ◽  
Lactate Dehydrogenase ◽  
Acid Production ◽  
Lactic Acid Production ◽  
Water Electrolysis ◽  
Rhodococcus Opacus ◽  
Beta Alanine ◽  
Polymer Precursors ◽  
Autotrophic Bacteria

AbstractHydrogen oxidizing autotrophic bacteria are promising hosts for CO2 conversion into chemicals. In this work, we engineered the metabolically versatile lithoautotrophic bacterium Rhodococcus opacus strain DSM 43205 for synthesis of polymer precursors. Aspartate decarboxylase (panD) or lactate dehydrogenase (ldh) were expressed for beta-alanine or L-lactic acid production, respectively. The heterotrophic cultivations on glucose produced 25 mg L-1 beta-alanine and 742 mg L-1 L-lactic acid, while autotrophic cultivations with CO2, H2 and O2 resulted in the production of 1.8 mg L-1 beta-alanine and 146 mg L-1 L-lactic acid. Beta-alanine was also produced at 345 µg L-1 from CO2 in electrobioreactors, where H2 and O2 were provided by water electrolysis. This work demonstrates that R. opacus DSM 43205 can be readily engineered to produce chemicals from CO2 and provides base for its further metabolic engineering.

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