[Site-directed mutagenesis enhances the activity of benzylidene acetone synthase of polyketide synthase from Polygonum cuspidatum]. 2023

Zhimin He, and Wenrui Ma, and Liping Yu, and Heshu Lü, and Mingfeng Yang
Key Laboratory for Northern Urban Agriculture of Ministry of Agriculture and Rural Affairs, Beijing University of Agriculture, Beijing 102206, China.

Polygonum cuspidatum polyketide synthase 1 (PcPKS1) has the catalytic activity of chalcone synthase (CHS) and benzylidene acetone synthase (BAS), which can catalyze the production of polyketides naringenin chalcone and benzylidene acetone, and then catalyze the synthesis of flavonoids or benzylidene acetone. In this study, three amino acid sites (Thr133, Ser134, Ser33) that may affect the function of PcPKS1 were identified by analyzing the sequences of PcPKS1, the BAS from Rheum palmatum and the CHS from Arabidopsis thaliana, as well as the conformation of the catalytic site of the enzyme. Molecular modification of PcPKS1 was carried out by site-directed mutagenesis, and two mutants were successfully obtained. The in vitro enzymatic reactions were carried out, and the differences in activity were detected by high performance liquid chromatography (HPLC). Finally, mutants T133LS134A and S339V with bifunctional activity were obtained. In addition to bifunctional activities of BAS and CHS, the modified PcPKS1 had much higher BAS activity than that of the wild type PcPKS1 under the conditions of pH 7.0 and pH 9.0, respectively. It provides a theoretical basis for future use of PcPKS1 in genetic engineering to regulate the biosynthesis of flavonoids and raspberry ketones.

UI MeSH Term Description Entries
D005419 Flavonoids A group of phenyl benzopyrans named for having structures like FLAVONES. 2-Phenyl-Benzopyran,2-Phenyl-Chromene,Bioflavonoid,Bioflavonoids,Flavonoid,2-Phenyl-Benzopyrans,2-Phenyl-Chromenes,2 Phenyl Benzopyran,2 Phenyl Benzopyrans,2 Phenyl Chromene,2 Phenyl Chromenes
D000096 Acetone A colorless liquid used as a solvent and an antiseptic. It is one of the ketone bodies produced during ketoacidosis.
D000217 Acyltransferases Enzymes from the transferase class that catalyze the transfer of acyl groups from donor to acceptor, forming either esters or amides. (From Enzyme Nomenclature 1992) EC 2.3. Acyltransferase
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D016297 Mutagenesis, Site-Directed Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion. Mutagenesis, Oligonucleotide-Directed,Mutagenesis, Site-Specific,Oligonucleotide-Directed Mutagenesis,Site-Directed Mutagenesis,Site-Specific Mutagenesis,Mutageneses, Oligonucleotide-Directed,Mutageneses, Site-Directed,Mutageneses, Site-Specific,Mutagenesis, Oligonucleotide Directed,Mutagenesis, Site Directed,Mutagenesis, Site Specific,Oligonucleotide Directed Mutagenesis,Oligonucleotide-Directed Mutageneses,Site Directed Mutagenesis,Site Specific Mutagenesis,Site-Directed Mutageneses,Site-Specific Mutageneses
D045468 Fallopia japonica A plant species of the family POLYGONACEAE. Itadori tea is prepared from the root of this genus. Polygonum cuspidatum,Japanese Knotweed,Knotweed, Japanese,Fallopia japonicas,Japanese Knotweeds,Knotweeds, Japanese,Polygonum cuspidatums,cuspidatum, Polygonum,japonicas, Fallopia
D048630 Polyketide Synthases Large enzyme complexes composed of a number of component enzymes that are found in STREPTOMYCES which biosynthesize MACROLIDES and other polyketides. Polyketide Synthase,6-Deoxyerythronolide-B Synthase,Epothilone Polyketide Synthase,Erythromycin Polyketide Synthase,Griseusin Polyketide Synthase,Niddamycin Polyketide Synthase,Polyketide Synthase L1,Polyketide Synthase WA,Rifamycin Polyketide Synthase,Sterigmatocystin Polyketide Synthase,Type I Polyketide Synthase,Type II Polyketide Beta-Ketoacyl Synthase,Urdamycin Polyketide Synthase,WdPKS1 Protein,WhiE Polyketide Synthase,6 Deoxyerythronolide B Synthase,Polyketide Synthase, Epothilone,Polyketide Synthase, Erythromycin,Polyketide Synthase, Griseusin,Polyketide Synthase, Niddamycin,Polyketide Synthase, Rifamycin,Polyketide Synthase, Sterigmatocystin,Polyketide Synthase, Urdamycin,Polyketide Synthase, WhiE,Protein, WdPKS1,Synthase L1, Polyketide,Synthase WA, Polyketide,Synthase, 6-Deoxyerythronolide-B,Synthase, Epothilone Polyketide,Synthase, Erythromycin Polyketide,Synthase, Griseusin Polyketide,Synthase, Niddamycin Polyketide,Synthase, Polyketide,Synthase, Rifamycin Polyketide,Synthase, Sterigmatocystin Polyketide,Synthase, Urdamycin Polyketide,Synthase, WhiE Polyketide,Synthases, Polyketide,Type II Polyketide Beta Ketoacyl Synthase

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