Gene Editing Services for Ensifer adhaerens

Gene Editing Services for Ensifer adhaerens

1. Research Background

Ensifer adhaerens is an environmental microorganism with unique adhesion ability and broad metabolic potential, exhibiting great application prospects in agriculture, environmental remediation and synthetic biology. This strain can efficiently colonize soil particles, plant roots and biofilm interfaces. Its strong adhesive properties and versatile substrate degradation capability enable it to play vital roles in plant growth promotion, heavy metal passivation and organic pollutant degradation.

In recent years, with in-depth research on rhizosphere microbiome and bioremediation mechanisms, Ensifer adhaerens, as a model adhesive bacterium, has become a research hotspot for bacterial surface interaction, quorum sensing and synthetic biology engineering of non-model microorganisms.

Based on homologous recombination technology, we provide professional genome editing services for Ensifer adhaerens, including targeted gene knockout, large-fragment deletion, site-directed point mutation, and site-specific gene integration/overexpression. We can deliver verified positive mutant strains in multiple genetic backgrounds, aiming to offer efficient and reliable genetic engineering solutions for fundamental research and agricultural biotechnology development.

2. Strain Characteristics and Biological Background

(1) Gram staining characteristics Ensifer adhaerens is a Gram-negative bacterium with a typical cell wall structure consisting of an outer membrane and a thin peptidoglycan layer.

(2) Physiological and biochemical characteristicsThe cells are short rod-shaped, with peritrichous or subpolar flagella and strong motility. Its most prominent feature is the rich surface adhesion proteins and exopolysaccharides, which support firm attachment to various abiotic/biotic surfaces and stable biofilm formation. This strain is aerobic or facultatively anaerobic, with an optimal growth temperature of 28–30°C, and can maintain high metabolic activity under oligotrophic environments.

(3) Ecological and industrial significanceWidely distributed in soil, freshwater and plant rhizosphere, Ensifer adhaerens is capable of degrading organic pollutants such as aromatic hydrocarbons, pesticides and plastic additives. It also secretes siderophores, plant growth hormones (e.g., IAA) and ACC deaminase, which facilitate plant growth and abiotic stress tolerance. It possesses promising application value in agricultural microbial inoculants, soil bioremediation and biofilm regulation.

3. Homologous Recombination Gene Editing Technology

Homologous recombination is a precise gene editing strategy relying on the endogenous DNA repair system of microorganisms. It utilizes intrinsic homologous sequences to mediate the integration of exogenous DNA fragments, thereby achieving accurate modification of target genes. This technology supports targeted gene subcloning, insertion, knockout, replacement and site-directed point mutation.

Figure 1 Multiple target gene editing strategies based on homologous recombination

4. Core Application Areas

(1) Environmental bioremediationPrecisely knock out competitive metabolic branch genes in aromatic hydrocarbon degradation pathways (e.g., catechol 2,3-dioxygenase paralogs) to reduce the accumulation of toxic intermediates and enhance the degradation efficiency of polycyclic aromatic hydrocarbons, pesticides and plasticizers.

(2) Plant-microbe interaction researchIntegrate plant hormone synthesis gene clusters (e.g., iaaM, ipdC) or fluorescent marker genes (e.g., gfp, mCherry) into genomic safe harbors (e.g., attTn7 locus) to construct growth-promoting engineered strains for rhizosphere colonization. These strains can be applied to in situ monitoring of rhizosphere colonization dynamics and hormone secretion patterns.

(3) Biofilm and adhesion mechanism researchPrecisely modify genes encoding surface adhesion proteins (e.g., adrA, eps gene cluster) and c-di-GMP metabolic regulatory genes via point mutation and small-fragment deletion, to reveal the molecular regulatory network underlying biofilm formation on soil particles and plant root surfaces.

(4) Synthetic biology chassis strain constructionCombined with homologous recombination and CRISPR/Cas9 iterative editing, we systematically knock out non-essential metabolic bypass genes and competitive exopolysaccharide synthesis genes, to construct low-adhesion and high-efficiency protein-secretion chassis strains. Such strains can be used for heterologous biosynthesis of complex natural products and the development of environmental microbial biosensors.

5. Gene Editing Project Overview

5.1 Core Services

(1) Markerless gene knockoutTargeted gene knockout via professional homologous recombination editing system.

(2) Site-specific gene knock-inPrecise gene knock-in at designated genomic loci.

(3) Genomic gene overexpressionSite-specific integration and overexpression of target genes in the genome.

5.2 Technical Advantages

(1) High editing efficiencyEfficient target gene modification with rational antibiotic screening strategy.

(2) High targeting accuracyHomologous recombination ensures precise editing at designated genomic sites.

(3) One-stop full-process validationCovering experimental design, vector construction, bacterial transformation, mutant screening and genotypic identification.

6. Delivery Standards

(1) PCR amplification and Sanger sequencing validation reports of target loci

(2) Two cryovials of engineered strain glycerol stocks

(3) Transcription level detection data (for overexpression projects)

(4) Official project completion report

7. Case Study

We have long provided professional gene editing services for top universities, research institutions and biotechnology enterprises worldwide. A typical case is shown below:

Case: Genome Modification of Ensifer adhaerens

Project Content: Targeted replacement of specific functional genes in the Ensifer adhaerens genome.

Figure 2 Sequencing verification of successful target gene replacement

8. References

[1] Zhao, Y.X., et al. (2021). Biodegradation of flonicamid by Ensifer adhaerens CGMCC 6315 and enzymatic characterization of the nitrile hydratases involved. Microb Cell Fact.

[2] Mohammad Oves, et al. (2017). Ensifer adhaerens for heavy metal bioaccumulation, biosorption, and phosphate solubilization under metal stress condition. Journal of the Taiwan Institute of Chemical Engineers.

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