Engineering Escherichia coli biofilm to increase contact surface for shikimate and L-malate production

AbstractMicrobial organelles are a promising model to promote cellular functions for the production of high-value chemicals. However, the concentrations of enzymes and nanoparticles are limited by the contact surface in single Escherichia coli cells. Herein, the definition of contact surface is to improve the amylase and CdS nanoparticles concentration for enhancing the substrate starch and cofactor NADH utilization. In this study, two biofilm-based strategies were developed to improve the contact surface for the production of shikimate and L-malate. First, the contact surface of E. coli was improved by amylase self-assembly with a blue light-inducible biofilm-based SpyTag/SpyCatcher system. This system increased the glucose concentration by 20.7% and the starch-based shikimate titer to 50.96 g L−1, which showed the highest titer with starch as substrate. Then, the contact surface of E. coli was improved using a biofilm-based CdS-biohybrid system by light-driven system, which improved the NADH concentration by 83.3% and increased the NADH-dependent L-malate titer to 45.93 g L−1. Thus, the biofilm-based strategies can regulate cellular functions to increase the efficiency of microbial cell factories based on the optogenetics, light-driven, and metabolic engineering. Graphical Abstract

Download Full-text

Escherichia coli as a platform microbial host for systems metabolic engineering

Essays in Biochemistry ◽

10.1042/ebc20200172 ◽

2021 ◽

Author(s):

Dongsoo Yang ◽

Cindy Pricilia Surya Prabowo ◽

Hyunmin Eun ◽

Seon Young Park ◽

In Jin Cho ◽

...

Keyword(s):

Escherichia Coli ◽

Natural Products ◽

Metabolic Engineering ◽

Food Ingredients ◽

E Coli ◽

Renewable Biomass ◽

Microbial Cell Factories ◽

Cell Factories ◽

Systems Metabolic Engineering ◽

Microbial Host

Abstract Bio-based production of industrially important chemicals and materials from non-edible and renewable biomass has become increasingly important to resolve the urgent worldwide issues including climate change. Also, bio-based production, instead of chemical synthesis, of food ingredients and natural products has gained ever increasing interest for health benefits. Systems metabolic engineering allows more efficient development of microbial cell factories capable of sustainable, green, and human-friendly production of diverse chemicals and materials. Escherichia coli is unarguably the most widely employed host strain for the bio-based production of chemicals and materials. In the present paper, we review the tools and strategies employed for systems metabolic engineering of E. coli. Next, representative examples and strategies for the production of chemicals including biofuels, bulk and specialty chemicals, and natural products are discussed, followed by discussion on materials including polyhydroxyalkanoates (PHAs), proteins, and nanomaterials. Lastly, future perspectives and challenges remaining for systems metabolic engineering of E. coli are discussed.

Download Full-text

De novo Biosynthesis of Tyrosol Acetate and Hydroxytyrosol Acetate from Glucose in Engineered Escherichia Coli

10.21203/rs.3.rs-287007/v1 ◽

2021 ◽

Author(s):

Daoyi Guo ◽

Xiao Fu ◽

Yue Sun ◽

Xun Li ◽

Hong Pan

Keyword(s):

Escherichia Coli ◽

De Novo ◽

Virgin Olive Oil ◽

Carbon Sources ◽

Acetate Production ◽

E Coli ◽

Microbial Cell Factories ◽

Cell Factories ◽

Alcohol Acetyltransferase ◽

First Time

Abstract Background: Tyrosol and hydroxytyrosol derived from virgin olive oil and olives extract, have wide applications both as functional food components and as nutraceuticals. However, they have low bioavailability due to their low absorption and high metabolism in human liver and small intestine. Acetylation of tyrosol and hydroxytyrosol can effectively improve their bioavailability and thus increase their potential use in the food and cosmeceutical industries. There is no report on the bioproductin of tyrosol acetate and hydroxytyrosol acetate so far. Thus, it is of great significance to develop microbial cell factories for achieving tyrosol acetate or hydroxytyrosol acetate biosynthesis.Results: In this study, two de novo biosynthetic pathways for the production of tyrosol acetate and hydroxytyrosol acetate were constructed in Escherichia coli. First, an engineered E. coli that allows production of tyrosol from simple carbon sources was established. Four aldehyde reductases were compared, and it was found that yeaE is the best aldehyde reductase for tyrosol accumulation. Subsequently, the pathway was extended for tyrosol acetate production by further overexpression of alcohol acetyltransferase ATF1 for the conversion of tyrosol to tyrosol acetate. Finally, the pathway was further extended for hydroxytyrosol acetate production by overexpression of 4-hydroxyphenylacetate 3-hydroxylase HpaBC.Conclusion: We have successfully established the artificial biosynthetic pathway of tyrosol acetate and hydroxytyrosol acetate from fermentable sugars and demonstrated for the first time the direct fermentative production of tyrosol acetate and hydroxytyrosol acetate from glucose in engineered E. coli

Download Full-text

Reprogramming microbial populations using a programmed lysis system to improve chemical production

Nature Communications ◽

10.1038/s41467-021-27226-3 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Wenwen Diao ◽

Liang Guo ◽

Qiang Ding ◽

Cong Gao ◽

Guipeng Hu ◽

...

Keyword(s):

Escherichia Coli ◽

Division Of Labor ◽

Microbial Populations ◽

Chemical Production ◽

E Coli ◽

Microbial Cell Factories ◽

Cell Factories ◽

Phenotypic Structure ◽

Colicin M ◽

Butyrate Production

AbstractMicrobial populations are a promising model for achieving microbial cooperation to produce valuable chemicals. However, regulating the phenotypic structure of microbial populations remains challenging. In this study, a programmed lysis system (PLS) is developed to reprogram microbial cooperation to enhance chemical production. First, a colicin M -based lysis unit is constructed to lyse Escherichia coli. Then, a programmed switch, based on proteases, is designed to regulate the effective lysis unit time. Next, a PLS is constructed for chemical production by combining the lysis unit with a programmed switch. As a result, poly (lactate-co-3-hydroxybutyrate) production is switched from PLH synthesis to PLH release, and the content of free PLH is increased by 283%. Furthermore, butyrate production with E. coli consortia is switched from E. coli BUT003 to E. coli BUT004, thereby increasing butyrate production to 41.61 g/L. These results indicate the applicability of engineered microbial populations for improving the metabolic division of labor to increase the efficiency of microbial cell factories.

Download Full-text

Metabolic Engineering Escherichia coli for the Production of Lycopene

Molecules ◽

10.3390/molecules25143136 ◽

2020 ◽

Vol 25 (14) ◽

pp. 3136 ◽

Cited By ~ 2

Author(s):

Zhaobao Wang ◽

JingXin Sun ◽

Qun Yang ◽

Jianming Yang

Keyword(s):

Escherichia Coli ◽

Metabolic Engineering ◽

Regulatory Networks ◽

Carbon Sources ◽

Operation Technique ◽

Mva Pathway ◽

E Coli ◽

Microbial Cell Factories ◽

Cell Factories ◽

Lycopene Production

Lycopene, a potent antioxidant, has been widely used in the fields of pharmaceuticals, nutraceuticals, and cosmetics. However, the production of lycopene extracted from natural sources is far from meeting the demand. Consequently, synthetic biology and metabolic engineering have been employed to develop microbial cell factories for lycopene production. Due to the advantages of rapid growth, complete genetic background, and a reliable genetic operation technique, Escherichia coli has become the preferred host cell for microbial biochemicals production. In this review, the recent advances in biological lycopene production using engineered E. coli strains are summarized: First, modification of the endogenous MEP pathway and introduction of the heterogeneous MVA pathway for lycopene production are outlined. Second, the common challenges and strategies for lycopene biosynthesis are also presented, such as the optimization of other metabolic pathways, modulation of regulatory networks, and optimization of auxiliary carbon sources and the fermentation process. Finally, the future prospects for the improvement of lycopene biosynthesis are also discussed.

Download Full-text

Complete biosynthesis of a sulfated chondroitin in Escherichia coli

Nature Communications ◽

10.1038/s41467-021-21692-5 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Abinaya Badri ◽

Asher Williams ◽

Adeola Awofiranye ◽

Payel Datta ◽

Ke Xia ◽

...

Keyword(s):

Chondroitin Sulfate ◽

Scale Up ◽

Production Methods ◽

E Coli ◽

Microbial Cell Factories ◽

Cell Factories ◽

Microbial Product ◽

Dry Cell Weight ◽

One Step ◽

Dry Cell

AbstractSulfated glycosaminoglycans (GAGs) are a class of important biologics that are currently manufactured by extraction from animal tissues. Although such methods are unsustainable and prone to contamination, animal-free production methods have not emerged as competitive alternatives due to complexities in scale-up, requirement for multiple stages and cost of co-factors and purification. Here, we demonstrate the development of single microbial cell factories capable of complete, one-step biosynthesis of chondroitin sulfate (CS), a type of GAG. We engineer E. coli to produce all three required components for CS production–chondroitin, sulfate donor and sulfotransferase. In this way, we achieve intracellular CS production of ~27 μg/g dry-cell-weight with about 96% of the disaccharides sulfated. We further explore four different factors that can affect the sulfation levels of this microbial product. Overall, this is a demonstration of simple, one-step microbial production of a sulfated GAG and marks an important step in the animal-free production of these molecules.

Download Full-text

Genotypic and Phenotypic Traits That Distinguish Neonatal Meningitis-Associated Escherichia coli from Fecal E. coli Isolates of Healthy Human Hosts

Applied and Environmental Microbiology ◽

10.1128/aem.07869-11 ◽

2012 ◽

Vol 78 (16) ◽

pp. 5824-5830 ◽

Cited By ~ 35

Author(s):

Catherine M. Logue ◽

Curt Doetkott ◽

Paul Mangiamele ◽

Yvonne M. Wannemuehler ◽

Timothy J. Johnson ◽

...

Keyword(s):

Escherichia Coli ◽

Phylogenetic Group ◽

Neonatal Meningitis ◽

Phenotypic Traits ◽

Healthy Human ◽

Content Type ◽

Plasmid Content ◽

E Coli ◽

Healthy Humans ◽

Definition Of

ABSTRACTNeonatal meningitisEscherichia coli(NMEC) is one of the top causes of neonatal meningitis worldwide. Here, 85 NMEC and 204 fecalE. coliisolates from healthy humans (HFEC) were compared for possession of traits related to virulence, antimicrobial resistance, and plasmid content. This comparison was done to identify traits that typify NMEC and distinguish it from commensal strains to refine the definition of the NMEC subpathotype, identify traits that might contribute to NMEC pathogenesis, and facilitate choices of NMEC strains for future study. A large number ofE. colistrains from both groups were untypeable, with the most common serogroups occurring among NMEC being O18, followed by O83, O7, O12, and O1. NMEC strains were more likely than HFEC strains to be assigned to the B2 phylogenetic group. Few NMEC or HFEC strains were resistant to antimicrobials. Genes that best discriminated between NMEC and HFEC strains and that were present in more than 50% of NMEC isolates were mainly from extraintestinal pathogenicE. coligenomic and plasmid pathogenicity islands. Several of these defining traits had not previously been associated with NMEC pathogenesis, are of unknown function, and are plasmid located. Several genes that had been previously associated with NMEC virulence did not dominate among the NMEC isolates. These data suggest that there is much about NMEC virulence that is unknown and that there are pitfalls to studying single NMEC isolates to represent the entire subpathotype.

Download Full-text

Enteroaggregative Escherichia coli: epidemiology, virulence and detection

Journal of Medical Microbiology ◽

10.1099/jmm.0.46930-0 ◽

2007 ◽

Vol 56 (1) ◽

pp. 4-8 ◽

Cited By ~ 67

Author(s):

Andrej Weintraub

Keyword(s):

Escherichia Coli ◽

Tissue Culture ◽

Watery Diarrhoea ◽

Biological Assays ◽

E Coli ◽

The Past ◽

Sporadic Cases ◽

Trained Staff ◽

Definition Of ◽

Specific Virulence

Enteroaggregative Escherichia coli (EAEC) is a subgroup of diarrhoeagenic E. coli (DEC) that during the past decade has received increasing attention as a cause of watery diarrhoea, which is often persistent. EAEC have been isolated from children and adults worldwide. As well as sporadic cases, outbreaks of EAEC-caused diarrhoea have been described. The definition of EAEC is the ability of the micro-organism to adhere to epithelial cells such as HEp-2 in a very characteristic ‘stacked-brick’ pattern. Although many studies searching for specific virulence factor(s) unique for this category of DEC have been published it is still unknown why the EAEC cause persistent diarrhoea. In addition, the aggregative property of EAEC causes a lot of problems in serotyping due to the cells auto-agglutinating. The gold standard for identification of EAEC includes isolation of the agent and an adherence assay using tissue culture, viz. HEp-2 cells. This assay is in most cases reliable; however, emergence of ‘atypical’ EAEC has been described in several publications. In addition, the HEp-2 assay is time consuming, demands a tissue culture lab and trained staff. Several molecular biological assays have been described, however, none show 100 % specificity.

Download Full-text

Etude taxonomique d'entérobactéries appartenant ou apparentées à l'espèce Escherichia coli

Canadian Journal of Microbiology ◽

10.1139/m81-016 ◽

1981 ◽

Vol 27 (1) ◽

pp. 98-106 ◽

Cited By ~ 8

Author(s):

F. Gavini ◽

D. Izard ◽

P. A. Trinel ◽

B. Lefebvre ◽

H. Leclerc

Keyword(s):

Escherichia Coli ◽

Numerical Analysis ◽

Dna Hybridization ◽

Dna Relatedness ◽

E Coli ◽

Definition Of

Phenetic (numerical analysis) and genetic (DNA–DNA hybridization) studies were carried out on strains belonging or related to the species Escherichia coli. They have shown the diversity of its phenotypes, by the presence of plasmidic characters (citrate+, urease+, H2S+, tetrathionate reductase+, raffinose+, and saccharose+). New strains related phenetically to E. coli are also individualized. They showed less than 30% DNA relatedness with E. coli. A new definition of E. coli is presented.

Download Full-text

MazG, a Nucleoside Triphosphate Pyrophosphohydrolase, Interacts with Era, an Essential GTPase in Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.184.19.5323-5329.2002 ◽

2002 ◽

Vol 184 (19) ◽

pp. 5323-5329 ◽

Cited By ~ 58

Author(s):

Junjie Zhang ◽

Masayori Inouye

Keyword(s):

Escherichia Coli ◽

Direct Interaction ◽

Kanamycin Resistance ◽

Nucleoside Triphosphate ◽

Cellular Functions ◽

Genomic Libraries ◽

E Coli ◽

Yeast Two Hybrid System ◽

Nucleoside Triphosphate Pyrophosphohydrolase

ABSTRACT Era is an essential GTPase in Escherichia coli, and Era has been implicated in a number of cellular functions. Homologues of Era have been identified in various bacteria and some eukaryotes. Using the era gene as bait in the yeast two-hybrid system to screen E. coli genomic libraries, we discovered that Era interacts with MazG, a protein of unknown function which is highly conserved among bacteria. The direct interaction between Era and MazG was also confirmed in vitro, being stronger in the presence of GDP than in the presence of GTPγS. MazG was characterized as a nucleoside triphosphate pyrophosphohydrolase which can hydrolyze all eight of the canonical ribo- and deoxynucleoside triphosphates to their respective monophosphates and PPi, with a preference for deoxynucleotides. A mazG deletion strain of E. coli was constructed by replacing the mazG gene with a kanamycin resistance gene. Unlike mutT, a gene for another conserved nucleotide triphosphate pyrophosphohydrolase that functions as a mutator gene, the mazG deletion did not result in a mutator phenotype in E. coli.

Download Full-text