CN-121991917-A - Furanochromene polyketone Compound biosynthesis gene and application thereof

Abstract

The invention belongs to the fields of genetic engineering and biosynthesis, and in particular relates to biosynthesis of polyketides. The invention provides genes which have key effects on ring opening of modification groups after benzene rings in the biosynthesis of polyketides, and are named as sipD, sipE and sipH and encoded polypeptides thereof. Provides the application of the polypeptide in the biosynthesis of polyketides.

Inventors

• LV JIANMING
• HUANG JIAHUA
• WANG GAOQIAN
• LI SHAOYANG
• HU DAN
• GAO HAO

Assignees

• 暨南大学

Dates

Publication Date

20260508

Application Date

20260211

Claims (10)

1. 1. An isolated polypeptide or a composition comprising the same, characterized in that the polypeptide is selected from the amino acid sequences shown in at least one of SEQ ID NOs 1-3, 30, 34.
2. 2. An isolated polynucleotide or a composition comprising the same, wherein the polynucleotide encodes the polypeptide of claim 1, preferably a nucleotide sequence selected from at least one of SEQ ID NOs 4-6, 31, 35.
3. 3. A vector comprising the polynucleotide of claim 2.
4. 4. A recombinant cell comprising the vector of claim 3, said cell being a fungal cell, preferably selected from the group consisting of Simplicillium fungal, aspergillus oryzae cells, or a bacterial cell, preferably said bacterial cell being an e.
5. 5. A method for preparing a polyketide compound shown in a formula (I), which comprises the step of contacting a polypeptide or a composition containing the polypeptide or the composition according to claim 1, a vector according to claim 3 or a recombinant cell according to claim 4 with a compound of a orosequoyitol and/or a orosequoyitol acid, wherein the structures of the compound of the orosequoyitol and the compound of the formula (I) are shown in the following formula: 、 、 Wherein, the R 1 is selected from hydrogen, hydroxy, methyl, hydroxymethyl, methoxy; r 2 is selected from hydrogen, hydroxy, methyl, hydroxymethyl, carboxy; R 3 is selected from hydrogen, acetoxy, hydroxymethyl, aldehyde, carboxyl; r 4 is selected from hydrogen.
6. 6. The process according to claim 5, wherein the polypeptide of SEQ ID NO. 1 and/or SEQ ID NO. 2 is contacted with a compound of the type of sequoyins or derivatives thereof, or the polypeptide of SEQ ID NO.3 is contacted with a compound of the type of sequoyins or derivatives thereof, optionally using a peptide, composition, expression vector, recombinant cell comprising SEQ ID NO. 30 and/or SEQ ID NO. 34.
7. 7. The process for producing a polyketide of formula (I) according to claim 5 or 6, wherein the compound of formula (I) has the structure: 。
8. 8. Use of the polypeptide of claim 1, the polynucleotide of claim 2, the vector of claim 3 and/or the cell of claim 4 in the synthesis of polyketides having the structure: Wherein R 1 is selected from hydrogen, hydroxy, methyl, hydroxymethyl, methoxy, R 2 is selected from hydrogen, hydroxy, methyl, hydroxymethyl, carboxyl, R 3 is selected from hydrogen, acetoxy, hydroxymethyl, aldehyde, carboxyl, R 4 is selected from hydrogen.
9. 9. The use according to claim 8, characterized in that the compound of formula (I) is selected from 。
10. 10. A biosynthetic system for synthesizing polyketides of formula (I), said system comprising at least one of the following components (a) to (d): (a) The polypeptide or composition thereof of claim 1; (b) The polynucleotide of claim 2 or a composition thereof; (c) The vector of claim 3; (d) The recombinant cell of claim 4; Wherein the system further comprises a compound, oroxylin and/or oroxylin acid, or the system is configured to be able to produce a compound, oroxylin and/or oroxylin acid from an endogenous metabolic pathway of the host cell.

Description

Furanochromene polyketone Compound biosynthesis gene and application thereof Technical Field The invention belongs to the fields of genetic engineering and biosynthesis, and in particular relates to biosynthesis of polyketides. Background Formula (I) is a class of polyketides, which are classified as Xyloketal-type compounds due to their characteristic [2,3-b ] furan chromene parent nucleus. Such compounds are currently isolated only in fungi, and more than 20 have been found. Modern pharmacological research shows that the compound can obviously promote angiogenesis, reduce blood pressure, prevent and treat atherosclerosis, and has outstanding treatment effect in various cardiovascular and cerebrovascular disease models. As in atherosclerosis-molded mice, the representative compound xylokeal B exhibits greater anti-atherosclerosis capacity than the positive drug simvastatin. Therefore, the compound has remarkable treatment potential for cardiovascular and cerebrovascular diseases, and deserves further research and development. However, the yield of the compound in host bacteria is low, the construction of the compound skeleton multi-chiral center cannot be simply and efficiently completed by the existing chemical synthesis method, and the supply problem limits the deep development and utilization of the compound. The biosynthesis method is simple to operate, is environment-friendly, and has great application potential in the aspect of synthesizing the active compound with a complex structure. However, the biosynthesis of the fungal furan chromene type natural product is less studied at present, and the biosynthesis pathway of Xyloketal compounds is not reported. Therefore, the field is urgent to elucidate the biosynthesis path of Xyloketal compounds, and lays a foundation for the efficient synthesis of the compounds. Disclosure of Invention The invention aims to provide a biosynthesis gene of polyketide. The invention provides an isolated polypeptide or a composition comprising the same, which is characterized in that the polypeptide is selected from the amino acid sequences shown in at least one of SEQ ID NO 1-3, 30 and 34. In the sense that it may be a single polypeptide or a composition consisting of two or three polypeptides, said polypeptide or composition comprising it may preferably comprise the necessary excipients for the preparation of the formulation. Specifically, the enzyme gene provided by the invention comprises: SipD peptide (amino acid sequence shown as SEQ ID NO: 1), an O-methyltransferase, responsible for catalyzing the methylation modification of specific hydroxyl groups of the precursor sedge aldehyde, is a key to directing the flow direction of the synthesis pathway. SipE peptide (amino acid sequence is shown as SEQ ID NO: 2), a flavoprotein monooxygenase, can catalyze methylation intermediate to generate Baeyer-Villiger oxidation reaction, and the ring opening reaction is a core step for constructing a complex oxygen heterocyclic skeleton of a target compound. SipH peptide (amino acid sequence shown as SEQ ID NO: 3), a decarboxylase, is responsible for catalyzing the decarboxylation of the other branched precursor, and provides the necessary building block for subsequent polymerization. SipA peptide (amino acid sequence shown as SEQ ID NO: 30) and SipB peptide (amino acid sequence shown as SEQ ID NO: 34) are responsible for synthesizing the key precursor compound, sequoyins. SipA and SipB cooperate to catalyze the assembly and cyclization of polyketone skeleton to generate sequoyial as a reaction core, thus providing a starting material for subsequent modification and polymerization. The polypeptide has synergistic action in the biosynthesis of target polyketide, and the action mechanism and flow are as follows: the optional precursor supply step SipA with SipB first catalyzes the formation of the common starting precursor, the fermentation of the recombinant cells comprising SipA and/or SipB to form the sequoyins can be performed using fermentation substances, or the acid can be synthesized in vitro using the substrates acetyl-CoA, malonyl-CoA, and ATP and NADPH can be used to reduce the sequoyins to sequoyins. The branch path I (modification path) is that a part of sedge aldehyde is subjected to O-methylation under SipD catalysis to generate a methylation intermediate, and the intermediate is subjected to key Baeyer-Villiger oxidation reaction under SipE catalysis to form a key linear alpha, beta unsaturated ketone intermediate through subsequent endogenous reduction, hydrolysis, demethylation and decarboxylation steps. Branch two (decarboxylation path) another portion of the precursor, which may be produced by the same or related pathways, undergoes decarboxylation under the catalysis of SipH, the product of which may be further methylated by SipD to form another polymeric unit. And finally, carrying out spontaneous polymerization on the linear alpha, beta unsaturated ketone interm