Team:BUCT-China
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Research and Construction of Fatty Acids and Glyoxylic Acid Oscillators
Background
The GntR family is a class of genes with potential transcriptional regulatory functions located in the genome of a cell[1]. Many members of the GntR family have been shown to play an important role in the survival of various strains such as Escherichia coli, Vibrio cholerae, and Mycobacterium tuberculosis under various stressful conditions.[2]
Glyoxylic acid is an important organic chemical intermediate. It is widely used in the production of high value-added aromatic aldehydes such as vanillin and ethyl vanillin. It is also used in the production of biomodulator allantoin, herbicide glyphosate, and Glucosin, valerate and insecticide quetiapine; in medicine and food, glyoxylic acid is used in the production of antihypertensive drug alotrol, broad-spectrum antibiotic amoxicillin and feed additive calcium pantotheate; in addition, it also For the production of high-efficiency water treatment agent hydroxyphosphoric acid, in other fields, such as leather and electroplating applications are still in the development stage.[3]
The chemical synthesis of glyoxylic acid has been developed for many years, and the related research has not stopped. The popularity in the factory is higher, but the chemical synthesis method still brings many drawbacks, so the biosynthesis method has received more and more attention. The use of biological enzymes as catalysts to oxidize glycolic acid to produce glyoxylic acid is one of the most active methods for biosynthesis of glyoxylic acid in recent years. Based on this, the biosynthesis pathway of glyoxylic acid extracts highly active enzymes from plants, directly converts to glyoxylic acid, and then uses immobilized enzymes to produce them. The most potential is to construct high-yield genetically engineered bacteria. The glyoxylate operon designed by us can improve the efficiency of gene expression and provide a feasible research route and theoretical basis for the efficient expression of this operon gene, which is of great significance for constructing higher-yielding engineering bacteria in the future.
Description
The purpose of this experimental study was to study the key sequences, mechanism of action, and site of action of some transcriptional regulators in the GntR/FadR family. Based on this research, a new transcriptional regulatory factor was designed as a new biological component. And together with other components to construct a biological module or gene loop, through the dynamic regulation of the metabolic processes in the microbial cells, improve the yield and quality of the target product production.
The purpose of this project is to complete the research and construction of fatty acid and glyoxylate operons, enrich the IGEM biocomponent library, further utilize the completed operons, and combine the standard biological modules (Biobricks) in the component library to construct gene loops and establish An effective mathematical model that enables the prediction, manipulation, and measurement of sophisticated artificial biological systems..
Synthetic biology
Synthetic biology refers to the study of artificial biological systems based on genetic engineering and engineering methods of systems biology, from gene fragments, DNA molecules, gene regulatory networks and signal transduction pathways to the manual design and synthesis of cells, similar to modern integrated chemical engineering.[4] Apply engineering principles and methods to biotechnology fields such as genetic engineering and cell engineering. People connect "genes" into a network, allowing cells to accomplish various tasks envisioned by designers, "programming" cells, and synthetic biology. Limited to genetic engineering, genetic splicing, but the beginning of the construction of the genetic code, in order to use synthetic genetic factors to construct new organisms.
Research progress
We have obtained the complete sequence of fatty acid ec, glyoxylic acid, fatty acid bh operon and found a suitable substrate for fatty acid ec-hydroxy fatty acid, thus giving the fatty acid ec operon application value. On this basis, we carried out a series of treatments on the predicted glyoxylate operator sequence and fatty acid ec gene sequence, and successfully constructed a functionally complete glyoxylate operon and fatty acid ec operon.
Citation
[1]Jialing Hu,Lei Zhao,Min Yang. A GntR family transcription factor positively regulates mycobacterial isoniazid resistance by controlling the expression of a putative permease[J]. BMC Microbiology,2015,15(1).
[2]Zhiqiang Li,Shuli Wang,Hui Zhang,Jinliang Zhang,Li Xi,Junbo Zhang,Chuangfu Chen. Transcriptional regulator GntR of Brucella abortus regulates cytotoxicity, induces the secretion of inflammatory cytokines and affects expression of the type IV secretion system and quorum sensing system in macrophages[J]. World Journal of Microbiology and Biotechnology,2017,33(3).
[3]He Yuting. Research Progress in Biosynthesis of Glyoxylic Acid[J]. Life Science Research, 2003(s1):29-32
[4]Carmen McLeod,Brigitte Nerlich. Synthetic biology, metaphors and responsibility[J]. Life Sciences, Society and Policy,2017,13(1).