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Lignans hydroxylase

A technology of lignans and hydroxylation, applied in the direction of oxidoreductase, enzyme, enzyme, etc., can solve unclear problems

Inactive Publication Date: 2009-07-01
SUNTORY HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, (+)-Larretricin hydroxylase is a hydroxylase of lignans without oxygen at the 9 (9') position of the furan ring. Therefore, for the 9 Hydroxylase remains unclear

Method used

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Examples

Experimental program
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preparation example Construction

[0168] The preparation method (production method) of the transformant or cells is not particularly limited, for example, a method of introducing the above-mentioned recombinant vector into a host and then transforming it can be exemplified. In addition, the organisms to be transformed are not particularly limited, and may be various microorganisms, plants, or animals listed above for the host cells.

[0169] The transformant or cell according to the present invention is characterized in that the composition of lignans and / or hydroxylated lignans naturally present in the transformant or cell is changed. The transformant or cell involved in the present invention is preferably derived from a plant or its progeny or a tissue derived from a plant or its progeny, particularly preferably sesame, forsythia or flax. Such transformants or cells can increase or decrease the hydroxylated lignan content in lignan-producing organisms by using the method for controlling the hydroxylated lign...

Embodiment 1

[0234] (Example 1: Preparation of sesame cDNA library)

[0235] Total RNA was extracted from sesame seeds using the RNeasy Plant Mini Kit (Qiagen) according to the manufacturer's recommended method. Then, 5 µg of poly A(+) RNA was obtained using the Oligotex MAG mRNA Purification Kit (TaKaRa). cDNA libraries were prepared from poly A(+) RNA using the ZAP Express cDNA Synthesis Kit and Zap Express cDNA Gigapack 3 Gold Cloning Kit (Stratagene) according to the manufacturer's recommended method. The prepared library was 1 x 10 7 pfu / ml (reference: Ono et al., Proc. Natl. Acad. Sci. USA 103, 10116-10121.2006).

Embodiment 2

[0236] (Example 2: Preparation of Hybridization Probes)

[0237] After extracting total RNA from Arabidopsis thaliana using RNeasy Plant Mini Kit (QIAGEN), use SuperScript according to the conditions recommended by the manufacturer TM First—Strand Synthesis System for RT-PCR (Invitrogen), cDNA was synthesized from 1 μg of total RNA. The 2-oxoglutarate-dependent dioxygenase (hereinafter 20G-dioxygenase)-like gene At3g13610 of Arabidopsis was used At3g13610-Fw (sequence number 1) and At3g13610-Rv primers (sequence number 2), and The amplified fragments were used as screening probes.

[0238] Serial No. 1: At3g13610-Fw: 5'-ATG GCT CCA ACA CTC TTG ACA ACC CAA-3'

[0239] Serial No. 2: At3g13610-Rv: 5'-TCA GAT CTT GGC GTA ATC GAC GGT TTT-3'

[0240] A non-radioactive isotope DIG-nucleic acid detection system (Roche) was used according to the PCR conditions recommended by the manufacturer, and a DIG label was introduced into the fragments obtained by RT-PCR. Specifically, the c...

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Abstract

Disclosed is an enzyme having a lignan-hydroxylating activity, particularly an enzyme capable of catalyzing the reaction for transferring a hydroxy group into a lignan, preferably an enzyme capable of catalyzing the hydroxylation reaction from piperitol into 9-hydroxypiperitol or from pinoresinol into 9-hydroxypinoresinol. Specifically disclosed are: a polypeptide having a lignan-hydroxylating activity; a polynucleotide encoding the polypeptide; a vector or transformant carrying the polynucleotide; a method for producing a polypeptide having a lignan-hydroxylating activity by using the transformant; and others. The transformant having the polynucleotide introduced therein in an expressible manner is useful for the hydroxylation of a lignan or the production of a product by using lignan hydroxide in the fields of foods and various industries.

Description

technical field [0001] The present invention relates to an enzyme having hydroxyl group-introducing activity to lignans, and a method using the enzyme. Background technique [0002] Lignans are secondary metabolites of vascular plants (for example, sesamin and sesamolin, etc.), and are widely distributed in plants. So far, it has been reported that lignans are contained in seeds, fruits, cut ears, tubers and / or roots of plants, etc., and mainly act on the biological defense system in plants. Since this lignan has strong antioxidant properties, etc., it is attracting attention that it has physiological and pharmacological functions in a wide range other than plants. Lignans have the following structure, which is C 6 -C 3 The phenylpropanoid compound of the skeleton undergoes two molecular polymerization structures, and the 8,8' bonding type is the most common [see Lignans, D.C.Ayres and J.D.Loike (1990)]. [0003] Representative lignans include (+)-pinoresinol, (+)-sesami...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/11C12N15/63C12N5/10C12P17/18A01H1/00C07D493/04
CPCC12N15/8255A23L1/3053C12N9/0071C12N9/0004C07D493/04C12P17/181A23L33/18
Inventor 小野荣一郎冈田亚砂子福井祐子
Owner SUNTORY HLDG LTD
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