Efficiency Unleashed, Innovation Empowered with Zymomonas mobilis Engineering Service

Zymomonas mobilis is a parthenogenetic anaerobic Gram-negative bacillus with many unique physiological characteristics and excellent industrial production properties. It is the only microorganism known to be able to utilize the entner-doudoroff (ED) pathway under anaerobic conditions, and it has excellent properties such as high sugar uptake, ethanol yield, and ethanol tolerance. It has been emphasized in recent years as a cellular factory for bioenergy production from cellulosic biomass biorefineries. Thus, Zymomonas mobilis has great potential as a high-performing Bacterial Chassis Cell for the Production of biochemicals.

Quality Zymomonas mobilis Engineering Service

At CD BioGlyco, we conduct a range of research on Zymomonas mobilis including physiological, genetic, and metabolic engineering modifications, and provide a specialized Zymomonas mobilis engineering service to broaden its substrate utilization and product generation.

Fig.1 The process of Zymomonas mobilis engineering. (CD BioGlyco)

Genetic modification

When adapting Zymomonas mobilis, the two main approaches can be categorized as random and rational design.

• Random mutagenesis is a method of positive genetic modification. Specific methods include physical mutagenesis, chemical mutagenesis, transposon mutagenesis, and so on. We screen for specific mutants after mutagenesis.

  • Among them, transposable factors on suicide vectors can be used to mutate the Zymomonas mobilis chromosome and construct stable Zymomonas mobilis strains.

  • Chemical mutagens such as nitrosoguanidine, (NTG), caffeine, and ethyl methane sulfonate (EMS) can be used to mutate the Zymomonas mobilis genome and obtain strains with better properties.

• Rational design is the transfer of candidate biological components, logical circuits, and metabolic pathways into chassis host cells or direct modification of chassis cells based on cognition or conclusions inferred from learning from systems biology data, and is often achieved using methods such as plasmid overexpression or gene editing.

  • Expression of endogenous and exogenous genes in Zymomonas mobilis using vector plasmids is a common approach.

  • Multiple DNA fragments based on scarless ligation technology provide multi-gene, multi-pathway assembly.

  • Homologous recombination provides for the construction of multiple genetically inactivated strains.

  • Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas technology is a powerful tool for multiple genome modifications such as gene deletion and replacement, in situ modification, large fragment deletion, and simultaneous multiple gene editing.

Shuttle plasmid construction

• Plasmids derived from Escherichia coli and Zymomonas mobilis are integrated to obtain a shuttle plasmid that replicates in both Escherichia coli and Zymomonas mobilis, such that the plasmid exists and propagates in both hosts.

• A host-wide plasmid based on Gram-negative bacillus with replicons and mob genes that are all recognized within Gram-negative bacillus for transformation into Zymomonas mobilis using a splice transformation assay.

The above two plasmids contain replicons for replication in Escherichia coli/ Gram-negative bacillus and Zymomonas mobilis, specific resistance screening genes as well as enzymatic cleavage sites to efficiently construct the Zymomonas mobilis metabolic pathway.

Efficient Screening Platform construction

• Simultaneous characterization of the effects of multiple conditions on strain growth by establishing a micro growth monitoring system in Zymomonas mobilis.

• In combination with the use of fully automated screening technology, it can be used for rapid and automated screening of modified dominant strains.

• Application of biosensing-based high-throughput screening technology to Zymomonas mobilis enables isolation of candidate strains of Zymomonas mobilis.

Applications

• Zymomonas mobilis engineering can be widely used for screening of Zymomonas mobilis mutants and genetic libraries, optimization of fermentation conditions, and systems biology research.
• Zymomonas mobilis engineering can be used to explore biological components, logic circuits, metabolic pathways, and cell factory performance in genetics.
• Zymomonas mobilis engineering can be used to enhance the productivity of Zymomonas mobilis and expand the range of biochemicals produced from Zymomonas mobilis.

Advantages

• We combine various methods such as bioinformatics and machine learning to mine biological components from multi-omics data and provide biological services with a wide range of applications.
• We have first-class high-throughput assembly technology of metabolic pathways, integrate multi-omics data of Chassis Cells, and provide custom experimental solutions for clients.
• We provide clients with detailed interpretation of experimental reports and professional after-sales service.

At CD BioGlyco, we have advanced synthetic biology technology and first-class experimental platforms to provide quality Chassis Cell Development and research services to our clients. Our synthetic biology services have been well received by our clients. Please feel free to contact us if you would like to know more about synthetic biology.

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