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Methods of using O-methyltransferase for biosynthetic production of pterostilbene

A methyltransferase, biosynthesis technology, used in medicine and pharmacology industry, food field

Active Publication Date: 2016-11-09
CONAGEN INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present disclosure solves the technical problem of producing pterostilbene in a cell system such as yeast or bacteria

Method used

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  • Methods of using O-methyltransferase for biosynthetic production of pterostilbene
  • Methods of using O-methyltransferase for biosynthetic production of pterostilbene
  • Methods of using O-methyltransferase for biosynthetic production of pterostilbene

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Experimental program
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Embodiment 1

[0053] Construction of bacterial expression vectors

[0054] The PCR product of ROMT was cloned into pETite N-His SUMO Kan vector (Lucigen) according to the manufacturer's manual. The resulting plasmid with the right insert was confirmed by sequencing, Sumo-ROMT, and transformed into BL21(DE3) for heterologous gene expression.

[0055] To construct the 4CL::STS fusion gene, At4CL and VvSTS were fused using a PCR amplification strategy. The stop codon of 4CL was removed and a three amino acid linker (Gly-Ser-Gly) was introduced between the open reading frame of 4CL and the STS. This construction resulted in a 2.87 kb fusion gene construct encoding 4CL, tripeptide linker and STS. The fusion gene 4CL::STS cloned into Gateway entry vector using pCR8 / GW / TOPO TA cloning kit (Invitrogen) was transformed into single-use E. coli cells and then sequenced. The 4CL::STS fusion gene was amplified and cloned via BamHI / HindIII into the multiple cloning site of the pETDuet-1 vector, desi...

Embodiment 2

[0057] Construction of Yeast Expression Vectors

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Abstract

A biosynthetic method of making pterostilbene including expressing a 4- coumaratexoenzyme A ligase (4CL) in a cellular system, expressing a stilbene synthase (STS) in the cellular system, expressing a resveratrol O-methyltransferase (ROMT) in the cellular system, feeding p-coumaric acid to the cellular system, growing the cellular system in a medium, and producing pterostilbene.

Description

[0001] Cross References to Related Applications [0002] The present disclosure is a PCT patent application entitled Method of Using O-methyltransferase for Biosynthetic Production of Pterostilbene. This application claims priority to US Provisional Patent Application No. 61 / 898899, filed November 1, 2013, which is hereby incorporated by reference in its entirety. technical field [0003] The present disclosure is applicable to the food, medicine and pharmacology industries. The present disclosure generally relates to methods for the biosynthetic production of pterostilbene using O-methyltransferase (ROMT). Background technique [0004] Pterostilbene is a stilbene compound chemically related to resveratrol (resveratrol) and is found in blueberries and grapes. It belongs to a class of phytoalexins, agents produced by plants to fight infection. Based on animal studies, pterostilbene is believed to have anti-cancer, anti-hypercholesterolemic, anti-hypertriglyceridemic prope...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H9/00A01H11/00C12N15/52C12N5/04
CPCC12Y203/01095C12Y602/01012C12N9/1007C12N9/1029C12N9/93C12P7/22A61P25/28A61P31/00A61P35/00A61P3/06
Inventor 穆罕默德·瓦杜德·柏惠亚汪业春余晓丹
Owner CONAGEN INC
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