Gene participating in synthesis of cationic peptide compound and application thereof
A technology for cationic peptides and compounds, applied in the field of genes involved in the synthesis of cationic peptide compounds
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Embodiment 1
[0098] Example 1 Acquisition of genes involved in the synthesis of cationic peptide compounds
[0099] A group of genes involved in the synthesis of cationic peptide compounds can be defined as containing 11 genes. These 11 genes are thioesterase protein gene, polyketide synthase gene, ketoreductase gene, non-ribosomal peptide synthetase gene (NRPS gene) (6 kinds), L-threonine-3-dehydrogenase gene , 2-amino-3-ketobutyrate CoA ligase gene.
[0100] Among the above-mentioned group of genes, the enzyme encoded by the NRPS gene has the function of forming the basic skeleton of the cationic peptide compound. That is, the enzyme forms a peptide backbone consisting of 13 amino acids, a fatty acid, and an aminoalcohol: [3-methyl-2-oxopentanoic acid (α-KIL)-dehydrothreonine (Thr)-methionine Acid (Met) - Ornithine (Orn) - Isoleucine (Ile) - Valine (Val) - Valine (Val) - Lysine (Lys) - Valine (Val) - Leucine acid (Leu)-lysine (Lys)-tyrosine (Tyr)-leucine (Leu)-valine (Val)]. The enzy...
Embodiment 2
[0136] Example 2 Functional evaluation of domains in each module
[0137] When the first A domain differs from the amino acid sequence of SEQ ID NO: 1, whether it can function as the A domain corresponding to 3-methyl-2-oxopentanoic acid can be evaluated as follows.
[0138] First, a mutant gene was designed to encode a first mutant A domain designed to be different from the amino acid sequence of SEQ ID NO:1. This mutant gene is expressed in an appropriate host, and it is confirmed whether a compound having the basic peptide skeleton of the above cationic peptide compound is synthesized in the host or in the metabolites in the culture supernatant. If among the metabolites there is a compound with the basic peptide backbone for the synthesis of the above-mentioned cationic peptide compounds, the designed first mutant A domain can be evaluated to function as the A domain corresponding to 3-methyl-2-oxopentanoic acid. function. Likewise, when the second A domain to the fourtee...
Embodiment 3
[0155] Example 3 Analysis of Gene Domain Structure
[0156] This example uses the antiSMASH program to analyze the domain structure of the protein (SEQ ID NO: 48-60) encoded by the gene involved in the synthesis of cationic peptide compounds of the present invention. See figure 2 ,From figure 2 It can be seen from the figure that the database in the antiSMASH program contains the structural domains in the protein encoded by the gene involved in the synthesis of cationic peptide compounds of the present invention.
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