A kind of method utilizing trans-anethole oxygenase to synthesize anisaldehyde

A technology of trans-anethole and brain oxygenase, which is applied in the field of preparation of anisaldehyde to achieve mild reaction conditions, reduce pollution, and broaden the range of enzyme sources

Active Publication Date: 2021-07-16
YULIN NORMAL UNIVERSITY
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The trans-anethole oxygenase of Pseudomonas putida JYR-1 is the only example that has been isolated so far, and there are no related enzymes reported in other strains that can use trans-anethole as the sole carbon source

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of method utilizing trans-anethole oxygenase to synthesize anisaldehyde
  • A kind of method utilizing trans-anethole oxygenase to synthesize anisaldehyde
  • A kind of method utilizing trans-anethole oxygenase to synthesize anisaldehyde

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) According to the amino acid sequence of CFIO01_02901 in the complete genome sequence information of the plant pathogen Colletotrichum fioriniae, a nucleotide sequence conforming to the codon preference of Escherichia coli was designed, the start codon was ATG, and the stop codon was ATG It is TAA, the length of the ORF is 993bp, and it is named as tao gene. Complete sequence synthesis of the tao gene (commissioned by Sangon Bioengineering Shanghai (Co., Ltd.)). The amino acid sequence of trans-anethole oxygenase from Colletotrichum fioriniae has 33% identity with that of Pseudomonas putida JYR-1.

[0038] (2) In order to place the ORF between SpeI and XhoI of pET-41a, a small sequence of TCAACTAGTGGTTCTGGT was added in front of the 5' end of the ORF, and CTCGAGCAC was added after the 3' end of the ORF, and a 1020bp fragment was commissioned by Shanghai Sangong ( 993+18+9=1020bp), cut with SpeI and XhoI and inserted into pET-41a(+) (purchased from Novagen) between S...

Embodiment 2

[0047] Example 2 Functional verification of the recombinant vector pET41-tao of the oxygenase gene tao

[0048] After the expression was induced in step (3) of Example 1, the bacterial solution was centrifuged at 10,000 r / min and 4° C. for 10 min to collect the bacterial cells, resuspended with basic medium, washed and centrifuged. The basic medium formula is: 1.2g (NH 4 ) 2 SO 4 , 0.1g CaCl 2 ·H 2 O, 0.1g MgSO 4 ·7H 2 O, 0.01g FeSO 4 ·7H 2 O, 0.2g K 2 HPO 4 , 0.1g KH 2 PO 4 and 1200mL distilled water. Wash once more with minimal medium. Add appropriate amount of basic medium and 0.5mmol / L glucose to make OD 600 =2.0, then add 1mmol / L trans-anethole, and react for 24 hours under the conditions of 25°C and 250r / min stirring speed to generate anisaldehyde. Extract with an equal volume of ethyl acetate for more than 30 minutes, collect the organic phase, and use it for the determination of anisaldehyde. The result shows that 0.7 mmol / L anisaldehyde is produced. It...

Embodiment 3

[0050] The difference between this example and Example 1 is that the reaction condition of step (7) is: react at 25° C. for 1 hour.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention discloses a method for synthesizing anisaldehyde by using trans-anethole oxygenase, which comprises the following steps: (1) constructing an engineering strain containing a nucleotide sequence encoding trans-anethole oxygenase by using a genetic engineering method; The nucleotide sequence of the trans-anethole oxygenase is derived from pine needle anthracnose, and the nucleotide sequence is shown in SEQ ID NO.1; (2) using the engineering strain to cultivate and separate and purify to obtain the trans-anethole oxygenase (3) using trans-anethole oxygenase to catalyze the oxygenation reaction of trans-anethole to obtain anisaldehyde. The invention uses enzymes to catalyze trans-anethole to generate anisaldehyde, the reaction conditions are mild, the environmental pollution caused by the reaction system is reduced, and the obtained products can be classified as natural fragrances.

Description

technical field [0001] The invention relates to a method for preparing anisaldehyde, in particular to a method for synthesizing anisaldehyde by trans-anethole oxygenase. Background technique [0002] Star anise oil, also known as star anise oil and fennel oil, is an aromatic essential oil extracted from star anise fruit or branches and leaves. It is a colorless, transparent or light yellow oily liquid at room temperature. special fragrance. Star anise oil contains 80-90% trans-anethole. The chemical name of trans-anethole is p-propenylanisole, and the molecular formula is C 10 h 12 O, the molecular weight is 148.2. Anisaldehyde (p-anisaldehyde, anisaldehyde) is also known as p-methoxybenzaldehyde, 4-methoxybenzaldehyde, p-anisaldehyde, anisaldehyde, and its molecular formula is C 8 h 8 o 2 , with a long-lasting hawthorn aroma. It can be used as the main fragrance in hawthorn flower, sunflower and lilac essence; it can be used as fragrance agent in lily of the valley ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/24
CPCC12P7/24
Inventor 林谦
Owner YULIN NORMAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap