Reduction of lactate production in Lactococcus lactis, a combined strategy: metabolic engineering by introducing foreign alanine dehydrogenase gene and hemin addition

2006 ◽  
Vol 23 (7) ◽  
pp. 947-953 ◽  
Author(s):  
Agustin Krisna Wardani ◽  
Keisuke Nagahisa ◽  
Hiroshi Shimizu ◽  
Suteaki Shioya
Keyword(s):  
Metabolic Engineering ◽  
Lactococcus Lactis ◽  
Lactate Production ◽  
Alanine Dehydrogenase ◽  
Dehydrogenase Gene ◽  
Combined Strategy

scholarly journals Metabolic Engineering of Mannitol Production in Lactococcus lactis: Influence of Overexpression of Mannitol 1-Phosphate Dehydrogenase in Different Genetic Backgrounds

2004 ◽  
Vol 70 (7) ◽  
pp. 4286-4292 ◽  
Author(s):  
H. Wouter Wisselink ◽  
Astrid E. Mars ◽  
Pieter van der Meer ◽  
Gerrit Eggink ◽  
Jeroen Hugenholtz
Keyword(s):  
Metabolic Engineering ◽  
Lactococcus Lactis ◽  
High Performance ◽  
Nuclear Magnetic Resonance Analysis ◽  
Resting Cells ◽  
Resonance Analysis ◽  
Glucose Depletion ◽  
Dehydrogenase Gene ◽  
Mannitol Production ◽  
Phosphofructokinase Activity

ABSTRACT To obtain a mannitol-producing Lactococcus lactis strain, the mannitol 1-phosphate dehydrogenase gene (mtlD) from Lactobacillus plantarum was overexpressed in a wild-type strain, a lactate dehydrogenase(LDH)-deficient strain, and a strain with reduced phosphofructokinase activity. High-performance liquid chromatography and 13C nuclear magnetic resonance analysis revealed that small amounts (<1%) of mannitol were formed by growing cells of mtlD-overexpressing LDH-deficient and phosphofructokinase-reduced strains, whereas resting cells of the LDH-deficient transformant converted 25% of glucose into mannitol. Moreover, the formed mannitol was not reutilized upon glucose depletion. Of the metabolic-engineering strategies investigated in this work, mtlD-overexpressing LDH-deficient L. lactis seemed to be the most promising strain for mannitol production.

scholarly journals Overproduction of Heterologous Mannitol 1-Phosphatase: a Key Factor for Engineering Mannitol Production by Lactococcus lactis

2005 ◽  
Vol 71 (3) ◽  
pp. 1507-1514 ◽  
Author(s):  
H. Wouter Wisselink ◽  
Antoine P. H. A. Moers ◽  
Astrid E. Mars ◽  
Marcel H. N. Hoefnagel ◽  
Willem M. de Vos ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Lactate Dehydrogenase ◽  
Lactococcus Lactis ◽  
Clear Correlation ◽  
Dehydrogenase Gene ◽  
Mannitol Production ◽  
Key Factor ◽  
Phosphofructokinase Activity ◽  
Overexpression System ◽  
Substrate Concentrations

ABSTRACT To achieve high mannitol production by Lactococcus lactis, the mannitol 1-phosphatase gene of Eimeria tenella and the mannitol 1-phosphate dehydrogenase gene mtlD of Lactobacillus plantarum were cloned in the nisin-dependent L. lactis NICE overexpression system. As predicted by a kinetic L. lactis glycolysis model, increase in mannitol 1-phosphate dehydrogenase and mannitol 1-phosphatase activities resulted in increased mannitol production. Overexpression of both genes in growing cells resulted in glucose-mannitol conversions of 11, 21, and 27% by the L. lactis parental strain, a strain with reduced phosphofructokinase activity, and a lactate dehydrogenase-deficient strain, respectively. Improved induction conditions and increased substrate concentrations resulted in an even higher glucose-to-mannitol conversion of 50% by the lactate dehydrogenase-deficient L. lactis strain, close to the theoretical mannitol yield of 67%. Moreover, a clear correlation between mannitol 1-phosphatase activity and mannitol production was shown, demonstrating the usefulness of this metabolic engineering approach.

scholarly journals Metabolic engineering and synthetic biology employing Lactococcus lactis and Bacillus subtilis cell factories

2019 ◽  
Vol 59 ◽  
pp. 1-7 ◽  
Author(s):  
Amanda Y van Tilburg ◽  
Haojie Cao ◽  
Sjoerd B van der Meulen ◽  
Ana Solopova ◽  
Oscar P Kuipers
Keyword(s):  
Bacillus Subtilis ◽  
Metabolic Engineering ◽  
Synthetic Biology ◽  
Lactococcus Lactis ◽  
Subtilis Cell ◽  
Cell Factories

scholarly journals Redirecting Reductant Flux into Hydrogen Production via Metabolic Engineering of Fermentative Carbon Metabolism in a Cyanobacterium

2010 ◽  
Vol 76 (15) ◽  
pp. 5032-5038 ◽  
Author(s):  
Kelsey McNeely ◽  
Yu Xu ◽  
Nick Bennette ◽  
Donald A. Bryant ◽  
G. Charles Dismukes
Keyword(s):  
Metabolic Engineering ◽  
Carbon Metabolism ◽  
Anaerobic Metabolism ◽  
Lactate Production ◽  
Energy Conversion Efficiency ◽  
Liquid Chromatography Mass Spectrometry ◽  
Wild Type ◽  
Photoautotrophic Growth ◽  
Principal Source ◽  
Anaerobic Conversion

ABSTRACT Some aquatic microbial oxygenic photoautotrophs (AMOPs) make hydrogen (H2), a carbon-neutral, renewable product derived from water, in low yields during autofermentation (anaerobic metabolism) of intracellular carbohydrates previously stored during aerobic photosynthesis. We have constructed a mutant (the ldhA mutant) of the cyanobacterium Synechococcus sp. strain PCC 7002 lacking the enzyme for the NADH-dependent reduction of pyruvate to d-lactate, the major fermentative reductant sink in this AMOP. Both nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography-mass spectrometry (LC-MS) metabolomic methods have shown that autofermentation by the ldhA mutant resulted in no d-lactate production and higher concentrations of excreted acetate, alanine, succinate, and hydrogen (up to 5-fold) compared to that by the wild type. The measured intracellular NAD(P)(H) concentrations demonstrated that the NAD(P)H/NAD(P)+ ratio increased appreciably during autofermentation in the ldhA strain; we propose this to be the principal source of the observed increase in H2 production via an NADH-dependent, bidirectional [NiFe] hydrogenase. Despite the elevated NAD(P)H/NAD(P)+ ratio, no decrease was found in the rate of anaerobic conversion of stored carbohydrates. The measured energy conversion efficiency (ECE) from biomass (as glucose equivalents) converted to hydrogen in the ldhA mutant is 12%. Together with the unimpaired photoautotrophic growth of the ldhA mutant, these attributes reveal that metabolic engineering is an effective strategy to enhance H2 production in AMOPs without compromising viability.

scholarly journals High-Level Acetaldehyde Production in Lactococcus lactis by Metabolic Engineering

2005 ◽  
Vol 71 (2) ◽  
pp. 1109-1113 ◽  
Author(s):  
Roger S. Bongers ◽  
Marcel H. N. Hoefnagel ◽  
Michiel Kleerebezem
Keyword(s):  
Metabolic Engineering ◽  
Lactococcus Lactis ◽  
Zymomonas Mobilis ◽  
Nadh Oxidase ◽  
Pyruvate Decarboxylase ◽  
Resting Cells ◽  
Anaerobic Conditions ◽  
High Level ◽  
Acetaldehyde Production ◽  
Combined Overexpression

ABSTRACT Efficient conversion of glucose to acetaldehyde is achieved by nisin-controlled overexpression of Zymomonas mobilis pyruvate decarboxylase (pdc) and Lactococcus lactis NADH oxidase (nox) in L. lactis. In resting cells, almost 50% of the glucose consumed could be redirected towards acetaldehyde by combined overexpression of pdc and nox under anaerobic conditions.

scholarly journals In silico gene knockout prediction using a hybrid of Bat algorithm and minimization of metabolic adjustment

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mei Yen Man ◽  
Mohd Saberi Mohamad ◽  
Yee Wen Choon ◽  
Mohd Arfian Ismail
Keyword(s):  
Metabolic Engineering ◽  
Gene Knockout ◽  
Lactate Production ◽  
Bat Algorithm ◽  
Host Cells ◽  
Knock Out ◽  
Genetic Manipulations ◽  
Microbial Strains ◽  
Metabolic Adjustment ◽  
Minimization Of Metabolic Adjustment

Abstract Microorganisms commonly produce many high-demand industrial products like fuels, food, vitamins, and other chemicals. Microbial strains are the strains of microorganisms, which can be optimized to improve their technological properties through metabolic engineering. Metabolic engineering is the process of overcoming cellular regulation in order to achieve a desired product or to generate a new product that the host cells do not usually need to produce. The prediction of genetic manipulations such as gene knockout is part of metabolic engineering. Gene knockout can be used to optimize the microbial strains, such as to maximize the production rate of chemicals of interest. Metabolic and genetic engineering is important in producing the chemicals of interest as, without them, the product yields of many microorganisms are normally low. As a result, the aim of this paper is to propose a combination of the Bat algorithm and the minimization of metabolic adjustment (BATMOMA) to predict which genes to knock out in order to increase the succinate and lactate production rates in Escherichia coli (E. coli).

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