scholarly journals Escherichia coli Strains Engineered for Homofermentative Production of d-Lactic Acid from Glycerol

2010 ◽  
Vol 76 (13) ◽  
pp. 4327-4336 ◽  
Author(s):  
Suman Mazumdar ◽  
James M. Clomburg ◽  
Ramon Gonzalez
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactate Dehydrogenase ◽  
Cell Mass ◽  
Lactate Production ◽  
Fumarate Reductase ◽  
Glycerol Metabolism ◽  
E Coli ◽  
Fuels And Chemicals ◽  
Microaerobic Conditions

ABSTRACT Given its availability and low price, glycerol has become an ideal feedstock for the production of fuels and chemicals. We recently reported the pathways mediating the metabolism of glycerol in Escherichia coli under anaerobic and microaerobic conditions. In this work, we engineer E. coli for the efficient conversion of glycerol to d-lactic acid (d-lactate), a negligible product of glycerol metabolism in wild-type strains. A homofermentative route for d-lactate production was engineered by overexpressing pathways involved in the conversion of glycerol to this product and blocking those leading to the synthesis of competing by-products. The former included the overexpression of the enzymes involved in the conversion of glycerol to glycolytic intermediates (GlpK-GlpD and GldA-DHAK pathways) and the synthesis of d-lactate from pyruvate (d-lactate dehydrogenase). On the other hand, the synthesis of succinate, acetate, and ethanol was minimized through two strategies: (i) inactivation of pyruvate-formate lyase (ΔpflB) and fumarate reductase (ΔfrdA) (strain LA01) and (ii) inactivation of fumarate reductase (ΔfrdA), phosphate acetyltransferase (Δpta), and alcohol/acetaldehyde dehydrogenase (ΔadhE) (strain LA02). A mutation that blocked the aerobic d-lactate dehydrogenase (Δdld) also was introduced in both LA01 and LA02 to prevent the utilization of d-lactate. The most efficient strain (LA02Δdld, with GlpK-GlpD overexpressed) produced 32 g/liter of d-lactate from 40 g/liter of glycerol at a yield of 85% of the theoretical maximum and with a chiral purity higher than 99.9%. This strain exhibited maximum volumetric and specific productivities for d-lactate production of 1.5 g/liter/h and 1.25 g/g cell mass/h, respectively. The engineered homolactic route generates 1 to 2 mol of ATP per mol of d-lactate and is redox balanced, thus representing a viable metabolic pathway.

Homofermentative Production of d- orl-Lactate in Metabolically Engineered Escherichia coli RR1

1999 ◽  
Vol 65 (4) ◽  
pp. 1384-1389 ◽  
Author(s):  
Dong-Eun Chang ◽  
Heung-Chae Jung ◽  
Joon-Shick Rhee ◽  
Jae-Gu Pan
Keyword(s):  
Escherichia Coli ◽  
Lactate Dehydrogenase ◽  
Lactobacillus Casei ◽  
Lactate Production ◽  
Anaerobic Conditions ◽  
E Coli ◽  
Fermentation Product ◽  
Dehydrogenase Gene ◽  
Mutant Strains ◽  
Optically Pure

ABSTRACT We investigated metabolic engineering of fermentation pathways inEscherichia coli for production of optically pured- or l-lactate. Several pta mutant strains were examined, and a pta mutant of E. coli RR1 which was deficient in the phosphotransacetylase of the Pta-AckA pathway was found to metabolize glucose tod-lactate and to produce a small amount of succinate by-product under anaerobic conditions. An additional mutation inppc made the mutant produce d-lactate like a homofermentative lactic acid bacterium. When the pta ppcdouble mutant was grown to higher biomass concentrations under aerobic conditions before it shifted to the anaerobic phase ofd-lactate production, more than 62.2 g ofd-lactate per liter was produced in 60 h, and the volumetric productivity was 1.04 g/liter/h. To examine whether the blocked acetate flux could be reoriented to a nonindigenousl-lactate pathway, an l-lactate dehydrogenase gene from Lactobacillus casei was introduced into apta ldhA strain which lacked phosphotransacetylase andd-lactate dehydrogenase. This recombinant strain was able to metabolize glucose to l-lactate as the major fermentation product, and up to 45 g of l-lactate per liter was produced in 67 h. These results demonstrate that the central fermentation metabolism of E. coli can be reoriented to the production of d-lactate, an indigenous fermentation product, or to the production of l-lactate, a nonindigenous fermentation product.

In Vitro Inhibition of the Growth of Salmonella typhimurium and Escherichia coli 0157:H7 by Bacteria Isolated from the Cecal Contents of Adult Chickens

1991 ◽  
Vol 54 (7) ◽  
pp. 496-501 ◽  
Author(s):  
ARTHUR HINTON ◽  
GEORGE E. SPATES ◽  
DONALD E. CORRIER ◽  
MICHAEL E. HUME ◽  
JOHN R. DELOACH ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Salmonella Typhimurium ◽  
Mixed Cultures ◽  
Enterococcus Durans ◽  
E Coli ◽  
Pure Cultures ◽  
In Vitro Inhibition ◽  
Lactic Acids

A Veillonella species and Enterococcus durans were isolated from the cecal contents of adult broilers. Mixed cultures of Veillonella and E. durans inhibited the growth of Salmonella typhimurium and Escherichia coli 0157:H7 on media containing 2.5% lactose (w/v). The growth of S. typhimurium or E. coli 0157:H7 was not inhibited by mixed cultures containing Veillonella and E. durans on media containing only 0.25% lactose or by pure cultures of Veillonella or E. durans on media containing either 0.25% or 2.5% lactose. The mixed cultures of Veillonella and E. durans produced significantly (P<0.05) more acetic, propionic, and lactic acids in media containing 2.5% lactose than in media containing 0.25% lactose. The inhibition of the enteropathogens was related to the production of lactic acid from lactose by the E. durans and the production of acetic and propionic acids from lactic acid by the Veillonella.

Inactivation of Escherichia coli O157:H7, Salmonella enterica Serotype Enteritidis, and Listeria monocytogenes on Lettuce by Hydrogen Peroxide and Lactic Acid and by Hydrogen Peroxide with Mild Heat

2002 ◽  
Vol 65 (8) ◽  
pp. 1215-1220 ◽  
Author(s):  
CHIA-MIN LIN ◽  
SARAH S. MOON ◽  
MICHAEL P. DOYLE ◽  
KAY H. McWATTERS
Keyword(s):  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Lactic Acid ◽  
Listeria Monocytogenes ◽  
Salmonella Enterica ◽  
Salmonella Enteritidis ◽  
Sensory Characteristics ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Log Reduction

Iceberg lettuce is a major component in vegetable salad and has been associated with many outbreaks of foodborne illnesses. In this study, several combinations of lactic acid and hydrogen peroxide were tested to obtain effective antibacterial activity without adverse effects on sensory characteristics. A five-strain mixture of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis, and Listeria monocytogenes was inoculated separately onto fresh-cut lettuce leaves, which were later treated with 1.5% lactic acid plus 1.5% hydrogen peroxide (H2O2) at 40°C for 15 min, 1.5% lactic acid plus 2% H2O2 at 22°C for 5 min, and 2% H2O2 at 50°C for 60 or 90 s. Control lettuce leaves were treated with deionized water under the same conditions. A 4-log reduction was obtained for lettuce treated with the combinations of lactic acid and H2O2 for E. coli O157:H7 and Salmonella Enteritidis, and a 3-log reduction was obtained for L. monocytogenes. However, the sensory characteristics of lettuce were compromised by these treatments. The treatment of lettuce leaves with 2% H2O2 at 50°C was effective not only in reducing pathogenic bacteria but also in maintaining good sensory quality for up to 15 days. A ≤4-log reduction of E. coli O157:H7 and Salmonella Enteritidis was achieved with the 2% H2O2 treatment, whereas a 3-log reduction of L. monocytogenes was obtained. There was no significant difference (P > 0.05) between pathogen population reductions obtained with 2% H2O2 with 60- and 90-s exposure times. Hydrogen peroxide residue was undetectable (the minimum level of sensitivity was 2 ppm) on lettuce surfaces after the treated lettuce was rinsed with cold water and centrifuged with a salad spinner. Hence, the treatment of lettuce with 2% H2O2 at 50°C for 60 s is effective in initially reducing substantial populations of foodborne pathogens and maintaining high product quality.

scholarly journals Lactic Acid Bacteria (Lactobacillus bulgaricus and Streptococcus thermophillus) in Yogurt Inhibit the Growth of Escherichia coli, Salmonella typhimurium, and Shigella sp. In Vitro

2021 ◽  
Vol 31 (4) ◽  
pp. 2
Author(s):  
IDSAP Peramiarti
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Salmonella Typhimurium ◽  
Pathogenic Bacteria ◽  
Test Method ◽  
P Value ◽  
E Coli ◽  
Significant Difference

Diarrhea is defecation with a frequency more often than usual (three times or more) a day (10 mL/kg/day) with a soft or liquid consistency, even in the form of water alone. Pathogenic bacteria, such as Escherichia coli, Salmonella typhimurium, and Shigella sp., play a role in many cases, to which antibiotics are prescribed as the first-line therapy. However, since antibiotic resistance cases are often found, preventive therapies are needed, such as consuming yogurt, which is produced through a fermentation process by lactic acid bacteria (LAB). This research aimed to determine the activity of lactic acid bacteria (Liactobacillus bulgaricus and Streptococcus thermophilus) in yogurt in inhibiting the growth of the pathogenic bacteria E. coli, S. typhimurium, and Shigella sp. The research applied in vitro with the liquid dilution test method and the true experimental design research method with post-test-only and control group design. The design was used to see the inhibitory effect of yogurt LAB on the growth of E. coli, S. typhimurium, and Shigell sp. to compare the effect of several different yogurt concentrations, namely 20%, 40%, 60%, and 80%. The results of the Least Significance Different analysis showed that there was a significant difference between yogurt with a concentration of 0% and that with various concentrations in inhibiting the growth of E. coli, S. typhimurium, and Shigella sp. with a p-value of &lt;0.05. Whereas, there was no significant difference in the various concentrations of yogurt in inhibiting the growth of the three kinds of bacteria with a p-value of &gt; 0.05.<p class="Default" align="center"> </p>

scholarly journals Antimicrobial activity of lactic acid bacteria isolated from garri on Escherichia coli strains isolated from clinical and environmental samples

2020 ◽  
Vol 12 (4) ◽  
pp. 357-365
Author(s):  
H.I. Atta ◽  
A. Gimba ◽  
T. Bamgbose
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactobacillus Acidophilus ◽  
Plant Pathogens ◽  
Resistant Bacteria ◽  
Fermented Foods ◽  
Environmental Isolates ◽  
E Coli

Abstract. The production of bacteriocins by lactic acid bacteria affords them the ability to inhibit the growth of bacteria; they are particularly important in the biocontrol of human and plant pathogens. Lactic acid bacteria have been frequently isolated from fermented foods due to the high acidity these foods contain. In this study, lactic acid bacteria were isolated from garri, a popular Nigerian staple food, which is fermented from cassava, and their antagonistic activity against clinical and environmental isolates of Escherichia coli was determined. The species of Lactobacillus isolated include: Lactobacillus plantarum (50%), Lactobacillus fermentum (20%), Lactobacillus acidophilus (20%), and Lactobacillus salivarius (10%). Growth inhibition of the strains of E.coli was observed in Lactobacillus plantarum that inhibited the growth of both. The clinical and environmental isolates of E. coli were inhibited by Lactobacillus plantarum, while Lactobacillus acidophilus showed activity against only the clinical isolate. The greatest zone of inhibition against the strains of E. coli was recorded by Lactobacillus acidophilus (22.7±1.53 mm). The bacteriocins produced by Lactobacillus species have a good potential in the biocontrol of pathogens, and should be the focus of further studies on antibiotic resistant bacteria.

scholarly journals Fermentative Utilization of Glycerol by Escherichia coli and Its Implications for the Production of Fuels and Chemicals

2007 ◽  
Vol 74 (4) ◽  
pp. 1124-1135 ◽  
Author(s):  
Abhishek Murarka ◽  
Yandi Dharmadi ◽  
Syed Shams Yazdani ◽  
Ramon Gonzalez
Keyword(s):  
Escherichia Coli ◽  
Anaerobic Fermentation ◽  
Glycerol Metabolism ◽  
Nuclear Magnetic Resonance Analysis ◽  
Glycerol Fermentation ◽  
Glycerol Utilization ◽  
Negative Effect ◽  
Balance Analysis ◽  
Fuels And Chemicals ◽  
High Degree

ABSTRACT Availability, low prices, and a high degree of reduction make glycerol an ideal feedstock to produce reduced chemicals and fuels via anaerobic fermentation. Although glycerol metabolism in Escherichia coli had been thought to be restricted to respiratory conditions, we report here the utilization of this carbon source in the absence of electron acceptors. Cells grew fermentatively on glycerol and exhibited exponential growth at a maximum specific growth rate of 0.040 ± 0.003 h−1. The fermentative nature of glycerol metabolism was demonstrated through studies in which cell growth and glycerol utilization were observed despite blocking several respiratory processes. The incorporation of glycerol in cellular biomass was also investigated via nuclear magnetic resonance analysis of cultures in which either 50% U-13C-labeled or 100% unlabeled glycerol was used. These studies demonstrated that about 20% of the carbon incorporated into the protein fraction of biomass originated from glycerol. The use of U-13C-labeled glycerol also allowed the unambiguous identification of ethanol and succinic, acetic, and formic acids as the products of glycerol fermentation. The synthesis of ethanol was identified as a metabolic determinant of glycerol fermentation; this pathway fulfills energy requirements by generating, in a redox-balanced manner, 1 mol of ATP per mol of glycerol converted to ethanol. A fermentation balance analysis revealed an excellent closure of both carbon (∼95%) and redox (∼96%) balances. On the other hand, cultivation conditions that prevent H2 accumulation were shown to be an environmental determinant of glycerol fermentation. The negative effect of H2 is related to its metabolic recycling, which in turn generates an unfavorable internal redox state. The implications of our findings for the production of reduced chemicals and fuels were illustrated by coproducing ethanol plus formic acid and ethanol plus hydrogen from glycerol at yields approaching their theoretical maximum.

scholarly journals Microbiological Condition of Retail Beef Steaks: A Canadian Survey

2018 ◽  
Vol 7 (4) ◽  
pp. 1 ◽  
Author(s):  
Xianqin Yang ◽  
Julia Devos ◽  
Hui Wang ◽  
Mark Klassen
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Microbial Quality ◽  
Indicator Organisms ◽  
Mean Values ◽  
E Coli ◽  
Upper Limit ◽  
Brochothrix Thermosphacta ◽  
Beef Steaks

The second national baseline microbiological survey of beef steaks offered for retail in Canada was conducted in 2015. A total of 621 steaks of four types (cross rib, CR; inside round, IR; striploin, SL; top sirloin, TS) collected from 135 retail stores in five cities across Canada were tested. Swab samples each from swabbing the entire upper surface of each steak were processed for enumeration of seven groups of indicator organisms: total aerobes (AER), psychrotrophs (PSY), lactic acid bacteria (LAB), pseudomonads (PSE), Brochothrix thermosphacta (BRO), coliforms (COL) and Escherichia coli (ECO). The overall mean values (log CFU/100 cm2) were 5.17±1.29, 4.92±1.36, 4.79±1.42, 3.26±1.49, 2.34±1.88, and 0.80±1.05 for AER, PSY, LAB, PSE, BRO, and COL, respectively. ECO were not recovered from 87.3% of the steaks and when there was recovery, the numbers were mostly ≤ 1 log CFU/100 cm2. Strong correlation was found between the log numbers of AER and PSY, of AER and LAB, and of PSY and LAB, while the correlation between the log numbers of COL and ECO was weak. The numbers of COL and ECO from different groups of steak types or from different cities were not substantially different. Of the four types of steaks, IR had the lowest median values for AER, PSY, LAB, PSE and BRO, followed by CR. The microbiological condition of retail beef steaks in this survey was on par with that in the previous one, with very low numbers of generic E. coli being recovered from very few steaks and the indicators for microbial quality being at numbers much lower than the upper limit for shelf life of beef.

scholarly journals Properties of human testis-specific lactate dehydrogenase expressed from Escherichia coli

1991 ◽  
Vol 273 (3) ◽  
pp. 587-592 ◽  
Author(s):  
K M LeVan ◽  
E Goldberg
Keyword(s):  
Escherichia Coli ◽  
Lactate Dehydrogenase ◽  
Prokaryotic Expression ◽  
Specific Activity ◽  
Human Testis ◽  
Reading Frame ◽  
E Coli ◽  
Heat Stable ◽  
Prokaryotic Expression Vector ◽  
Tac Promoter

The cDNA encoding the C4 isoenzyme of lactate dehydrogenase (LDH-C4) was engineered for expression in Escherichia coli. The Ldh-c open reading frame was constructed as a cassette for production of the native protein. The modified Ldh-c cDNA was subcloned into the prokaryotic expression vector pKK223-3. Transformed E. coli cells were grown to mid-exponential phase, and induced with isopropyl beta-D-thiogalactopyranoside for positive regulation of the tac promoter. Induced cells expressed the 35 kDa subunit, which spontaneously formed the enzymically active 140 kDa tetramer. Human LDH-C4 was purified over 200-fold from litre cultures of cells by AMP and oxamate affinity chromatography to a specific activity of 106 units/mg. The enzyme was inhibited by pyruvate concentrations above 0.3 mM, had a Km for pyruvate of 0.03 mM, a turnover number (nmol of NADH oxidized/mol of LDH-C4 per min at 25 degrees C) of 14,000 and was heat-stable.

Influence of Sodium Chloride on Growth of Lactic Acid Bacteria and Subsequent Destruction of Escherichia coli O157:H7 during Processing of Lebanon Bologna

2001 ◽  
Vol 64 (8) ◽  
pp. 1145-1150 ◽  
Author(s):  
NAVEEN CHIKTHIMMAH ◽  
RAMASWAMY C. ANANTHESWARAN ◽  
ROBERT F. ROBERTS ◽  
EDWARD W. MILLS ◽  
STEPHEN J. KNABEL
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Low Ph ◽  
Escherichia Coli O157 ◽  
Quality Changes ◽  
E Coli ◽  
Fermented Product ◽  
Nacl Concentration ◽  
Different Levels

Due to undesirable quality changes, Lebanon bologna is often processed at temperatures that do not exceed 48.8°C (120°F). Therefore, it is important to study parameters that influence the destruction of Escherichia coli O157:H7 in Lebanon bologna. The objective of the present study was to determine the influence of curing salts (NaCl and NaNO2) on the destruction of E. coli O157:H7 during Lebanon bologna processing. Fermentation to pH 4.7 at 37.7°C reduced populations of E. coli O157:H7 by approximately 0.3 log10, either in the presence or absence of curing salts. Subsequent destruction of E. coli O157:H7 during heating of fermented product to 46.1°C was significantly reduced by the presence of 3.5% NaCl and 156 ppm NaNO2, compared to product without curing salts (P &lt; 0.01). The presence of a higher level of NaCl (5%) in Lebanon bologna inhibited the growth of lactic acid bacteria (LAB), which yielded product with higher pH (~5.0) and significantly reduced the destruction of E. coli O157:H7 even further (P &lt; 0.05). Lower concentrations of NaCl (0, 2.5%) yielded Lebanon bologna with higher LAB counts and lower pHs, compared to product with 5% NaCl. When lactic acid was used to adjust pH in product containing different levels of NaCl, it was determined that low pH was directly influencing destruction of E. coli O157:H7, not NaCl concentration.

pH-mediated control of anti-aggregation activities of cyanobacterial and E. coli chaperonin GroELs

2020 ◽  
Author(s):  
Tahmina Akter ◽  
Hitoshi Nakamoto
Keyword(s):  
Escherichia Coli ◽  
Lactate Dehydrogenase ◽  
Malate Dehydrogenase ◽  
Citrate Synthase ◽  
Ph Dependence ◽  
Chaperone Activity ◽  
Ph Change ◽  
Aggregation Assay ◽  
E Coli ◽  
Aggregation Activity

Abstract In contrast to Escherichia coli, cyanobacteria have multiple GroELs, the bacterial homologues of chaperonin/Hsp60. We have shown that cyanobacterial GroELs are mutually distinct and different from E. coli GroEL with which the paradigm for chaperonin structure/function has been established. However, little is known about regulation of cyanobacterial GroELs. This study investigated effect of pH (varied from 7.0 to 8.5) on chaperone activity of GroEL1 and GroEL2 from the cyanobacterium Synechococcus elongatus PCC7942 and E. coli GroEL. GroEL1 and GroEL2 showed pH dependency in suppression of aggregation of heat-denatured malate dehydrogenase, lactate dehydrogenase and citrate synthase. They exhibited higher anti-aggregation activity at more alkaline pHs. Escherichia coli GroEL showed a similar pH-dependence in suppressing aggregation of heat-denatured lactate dehydrogenase. No pH dependence was observed in all the GroELs when urea-denatured lactate dehydrogenase was used for anti-aggregation assay, suggesting that the pH-dependence is related to some denatured structures. There was no significant influence of pH on the chaperone activity of all the GroELs to promote refolding of heat-denatured malate dehydrogenase. It is known that pH in cyanobacterial cytoplasm increases by one pH unit following a shift from darkness to light, suggesting that the pH-change modulates chaperone activity of cyanobacterial GroEL1 and GroEL2.

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