Research Terms
This genetically modified cyanobacterial host system uses Synechocystis as the host bacteria to produce structurally diverse natural products. Cyanobacterial natural products and their analogs are useful chemicals in health, agriculture and other industries. While some available cyanobacterial strains have been employed in producing valuable chemicals, e.g., biofuels, commodity chemicals, and biomaterials, the yield of natural products from native cyanobacteria is often low. Additionally, a problem exists to produce these chemicals in commonly used bacterial systems because they can have difficulty reading the inserted cyanobacterial genetic material. In this regard, the use of genetically modified cyanobacterial systems for the production of cyanobacterial natural products is enticing, but is still in its infancy.
Researchers at the University of Florida have developed a cyanobacterial system that efficiently reads the inserted genetic material and generates a high yield of the target products and its analogs, providing a variety of potentially viable compounds for research and development or commercial applications.
An easily customizable engineered cyanobacterial system that photosynthetically produces diverse chemicals for a variety of applications such as biofuels, commodity chemicals, biomaterials and active ingredients
The cyanobacterial system uses the cyanobacterium Synechocystis as the host to produce bioactive chemicals. Synechocystis is genetically modified to produce the target natural products and analogs. Cyanobacterial systems economically produce compounds since they only need sunlight, carbon dioxide, and water for growth and chemical production. The cyanobacterium can be engineered with gene clusters to photosynthetically overproduce compounds of interest. Proof of principle of this technology was demonstrated by engineering Synechocystis as a host to produce the sunscreen ingredient shinorine. The target genes responsible for shinorine synthesis were isolated from the filamentous cyanobacterium Fischerella which is a native producer of shinorine. The engineered strains produce ten-fold more shinorine than the native cyanobacterium.
This drug platform employs living multicellular assemblies that can deliver drug molecules in controlled patterns to specific tissues in order to perform complex therapeutic functions. Unlike standard small-molecule drugs and biopharmaceuticals, living drugs are therapeutic systems composed of living cells, which are able to perform medical functions. Most living drugs use genetically engineered cells of a single type, as with CAR T cell therapy, which has been successful in treating certain cancers, and stem cell-based therapies, which have treated other diseases effectively. However, relying on just one cell limits the potential functionality of living drugs.
Researchers at the University of Florida have developed a system to create multicellular living drugs made up of two or more types of alive cells (i.e., drug-secreting and supporting cells) that can work together to perform complex therapeutic functions. The choice of alive cells includes genetically programmed prokaryotic cells, eukaryotic cells, or a mixture of both. This new class of medicine employs a capsule that hydrolyzes in specific tissue areas to release the multiple types of cells selectively, controlling dose and duration. The multiple types of cells can perform various functions, including secreting specific drug molecules in programmed combinations or sensing a disease via associated biomarkers.
New class of multicellular living drugs that increase retention, viability, and drug-secreting efficacy in targeted host tissues
Multicellular living drugs are isolated micro-ecosystems comprised of nano-bio-engineered assemblies of living cells, which include three major components to act as medicines. A cross-linked polymer boundary shell provides protection for the cells and controls the uptake of nutrient molecules. Functional cells within the shell are programmed to perform a variety of tasks such as synthesizing molecular drugs, assisting in metabolism, and transporting substances into the extracellular space. The third component is a collection of supporting cells, programmed genetic circuits that maintain homeostasis in the living drugs, regulate the release of products from functional cells, and eliminate the living drugs once their therapeutic purpose is over.
This nitration system produces large quantities of pharmaceutically relevant nitrated tryptophan and other nitroaromatics using biocatalysts. Nitrated tryptophan and other nitroaromatics are essential components of a wide variety of pharmaceuticals, including anti-cancer and anti-Parkinson drugs. The most common nitroaromatic, nitrobenzene, is valued at $9.3 billion globally, with a predicted increase in value of 5.6% from 2020 to 2027. Production of nitroaromatics classically requires environmentally hazardous nitric acid, creating safety concerns and generating large quantities of acidic waste. Advanced technologies enable nitration without nitric acid, but these nitration systems have not produced large enough quantities of nitroaromatics to be applicable in the pharmaceutical industry.
Researchers at the University of Florida have developed a nitration system that uses biocatalysts rather than nitric acid to produce large quantities of nitroaromatics at relatively low costs. This system is environmentally friendly and can be scaled to meet industrial pharmaceutical production requirements.
Efficient, industrial-scale production of nitroaromatics essential in pharmaceuticals using biocatalysts instead of environmentally-devastating nitric acid
The nitration system is a whole-cell biocatalytic process that uses an engineered strain of E. coli containing a nitration biocatalyst, a nitric oxide synthase, and a glucose dehydrogenase to produce nitro tryptophan and other complex chemicals in large quantities. This system eliminates the need for nitric acid used in traditional chemical nitration systems and improves on other biocatalytic nitration systems by using an engineered strain of E. coli that includes a nitric oxide synthase gene, significantly reducing production costs.
These biocatalysts can generate novel nitroaromatic end products and building blocks for the synthesis of more complex compounds in an environmentally friendly manner. Nitro aromatic and heterocyclic derivatives are important industrial chemicals that are used as food additives, pesticides, herbicides, polymers, dyes and pharmaceuticals. Small nitroaromatics are common building blocks of complex molecules in chemical synthesis and chemical aromatic nitration is widely used in organic synthesis. However, the current processes are not environmentally sound and have other challenges including poor selectivity, low yield, and generation of multiple isomers and by-products. Researchers at the University of Florida have developed a biocatalyst system that is capable of transferring a nitro group onto an indole moiety of a variety of tryptophan analogues with greater efficiency, a higher degree of selectivity, and minimal environmental impact.
Biocatalyst-based aromatic nitration reaction transfers the nitro functional group to the indole structural core motif with greater efficiency and a higher degree of selectivity than current processes. These biocatalysts can generate nitroaromatics in an environmentally friendly manner and this approach can be used for the generation of novel nitroaromatics.
This nitration process utilizes biocatalysts to add a nitro group to aromatic molecules. The biocatalysts are engineered as “self-sufficient enzymes,” meaning they are able to perform their function without adding any auxiliary redox proteins. Researchers create them by fusing the nitration-promoting P450 with reductase domains of irrelevant P450s from Bacillus megatorium and Rhodococcus species. The engineered self-sufficient cytochrome P450 enzymes are capable of nitrating the indole of L-tryptophan analogues carrying one or more substituents on the indole, using nitric oxide and oxygen as co-substrates.
The recombinant glycan-binding protein Ab-Y3, found in an edible, common mushroom, is useful for detecting, isolating, purifying, and screening proteins with specific N-glycans of medical significance, such as immunoglobins or IgG proteins. Glycan binding proteins are critical for a number of biological functions and are promising drug candidates and tools for the detection and treatment of cancer and many other diseases. Recombinant glycoprotein therapies are becoming increasingly common, including in applications to fight COVID-19, and the market for profiling and characterizing glycosylated molecules should reach $2.5 billion by 2030. Due to limitations in analytical techniques, high structural diversity, and rapid evolution of glycans, knowledge of glycan-binding proteins is generally limited to only a handful of proteins.
Researchers at the University of Florida have developed a system for producing a glycan-binding protein to detect, isolate, and purify certain N-glycosylated proteins. The glycan-binding protein detects unique glycosylation patterns and can screen for therapeutic proteins.
Glycan binding protein derived from low-cost mushrooms to isolate, purify, and screen for certain therapeutic proteins with N-glycans
This yeast expression system produces high yields of the glycan-binding protein, Agaricus bisporus lectin, occurring naturally in the common mushroom species, Agaricus bisporus. The produced glycan-binding protein has a unique binding specificity for glycans with Man3GlcNAc2 core structure and Gal-GlcNAc branches in human cell line-expressed recombinant proteins. Agaricus bisporus lectin can be useful for detecting, isolating, and purifying therapeutic proteins and antibodies with medically important complex N-glycans.
