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Method for producing nerol through microorganism fermentation

A technology of microbial fermentation and nerol, applied in biochemical equipment and methods, botany equipment and methods, fermentation, etc., can solve the problems of environmental pollution, high cost, consumption of petroleum resources, etc., achieve pollution reduction, easy transformation, The effect of fast metabolism

Active Publication Date: 2019-05-28
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, nerol is mainly synthesized by chemical method, but the current chemical synthesis method not only has high cost, but also causes environmental pollution and consumes precious petroleum resources

Method used

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  • Method for producing nerol through microorganism fermentation
  • Method for producing nerol through microorganism fermentation
  • Method for producing nerol through microorganism fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method utilizing microbial fermentation to produce nerol, comprising the steps of:

[0034] 1) Synthesize the genes NDPS1 and GmNES encoding neryl pyrophosphate synthase and nerol synthase:

[0035] The genes NDPS1 (Solanum lycopersicumND PS1 gene sequence shown in SEQ ID NO.1) and GmNES (Glycine max GmNES gene sequence shown in SEQ ID NO.3) encoding neroliyl pyrophosphate synthase and nerol synthase were provided by Nanjing Synthesized by Jinweizhi Company.

[0036] Using the synthesized GmNES gene as a template, the GmNES gene fragment was extended by PCR using the following primers:

[0037] GmNES upstream primer: CTGGTGCCGCGCGGCAGCCATATGGACAACATCTCATCAA

[0038] GmNES downstream primer: TCCACCAGTCATGCTAGCCATATTATTCAATCACGAACTGCA

[0039] The amplified GmNES gene fragment was cloned into pET28a at the NdeI site by a seamless cloning method to obtain the plasmid pET28a-GmNES.

[0040] Using the synthesized NDPS1 gene as a template, the NDPS1 gene fragment was ex...

Embodiment 2

[0051] A method utilizing microbial fermentation to produce nerol, comprising the steps of:

[0052] 1) Acquisition of 3-hydroxy-3-methylglutaryl-CoA reductase gene tHMG1 and hydroxymethylglutaryl-CoA synthetase gene ERG13:

[0053] Using the genome of Saccharomyces cerevisiae as a template, each gene fragment was extended by PCR using the following primers, and the restriction sites are underlined:

[0054] tHMG1 upstream primer: CCGCTCTAGAACTAGTGGATCAATAAGGAGATATACCATGGACCAATTGGTGAAAAC

[0055] tHMG1 downstream primer: GAATTCCTGCAGCCCGGGGGATCCTTAGGATTTAATGCAGGTGA

[0056] ERG13 upstream primer: CTGCATTAAATCCTAAGGATCAATAAGGAGATATACCATGAAACTCTCAACTAAACT

[0057] ERG13 downstream primer: GAATTCCTGCAGCCCGGGGGATCCTTATTTTTTAACATCGTAAG.

[0058] 2) Acquisition of plasmid pLZ02:

[0059] The tHMG1 gene fragment amplified in step 1) was cloned into pBBR1mcs at the BamHI site by a seamless cloning method to obtain the plasmid pBBR1mcs-tHMG1, the schematic diagram of which is shown...

Embodiment 3

[0065] A method utilizing microbial fermentation to produce nerol, comprising the steps of:

[0066] 1) Acquisition of genes IDI1, MVD1, ERG8 and ERG12 of isopentenyl diphosphate isomerase, mevalonate pyrophosphate decarboxylase, phosphomevalonate kinase and mevalonate kinase:

[0067] Using the genome of Saccharomyces cerevisiae as a template, each gene fragment was extended by PCR using the following primers, and the restriction sites are underlined:

[0068] IDI1 upstream primer: ACTTTAAGAAGGAGATATACATATGACTGCCGACAACAATAGTA

[0069] IDI1 downstream primer: TCCACCAGTCATGCTAGCCATATTATAGCATTCTATGAATTTGC

[0070] MVD1 upstream primer: CAGCAAATGGGTCGCGGATCAATAAGGAGATATACCATGACCGTTTACACAGCATC

[0071] MVD1 downstream primer: TCGACGGAGCTCGAATTCGGATCCTTATTCCTTTGGTAGACCAG

[0072] EGR8 upstream primer: CTACCAAAGGAATAAGGATCAATAAGGAGATATACCATGTCAGAGTTGAGAGCCTT

[0073] ERG8 downstream primer: TCGACGGAGCTCGAATTCGGATCCTTATTTATCAAGATAAGTTT

[0074] ERG12 upstream primer: TATCTTGATAA...

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Abstract

The invention belongs to the field of production of renewable spices and foods, and chemical raw materials and in particular relates to a method for producing nerol through microorganism fermentation.The method comprises the following step: expressing neryl pyrophosphate synthase and nerol synthase genes in microorganisms to produce the nerol. By adopting the method provided by the invention, thenerol is directly synthesized through modifying the microorganisms and can be obtained, without the need of chemical synthesis, and the pollution to the environment is reduced; the method is a renewable, low-consumption and environment-friendly technology. The method for producing the nerol through the microorganisms can be directly applied to industrial production.

Description

technical field [0001] The invention belongs to the fields of renewable spices, food and chemical raw material production, and in particular relates to a method for producing nerol by microbial fermentation. Background technique [0002] Nerol, molecular formula C10H18O, molecular weight 154.25, is a monoterpene present in many essential oils such as lemongrass and hops, and was originally isolated from neroli oil. Neroliol has a pleasant aroma of roses and orange blossoms. The aroma is relatively mild, with a slight lemon-like fruity aroma. Neroliol is an isomer of geraniol, and its aroma is softer and more beautiful than geraniol. Clear with fresh notes of citrus and citrus. Nerol is a kind of precious spice with rose fragrance. It is used to prepare rose-type and orange blossom-type floral essences. It is widely used in the preparation of diet, food, and daily chemical high-end essences. It is also the synthesis of other important spices. It is an intermediate product a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C12N15/60C12N15/54C12N15/53C12N15/61C12P7/04
Inventor 柳志杰宗朕李冬生汪超高冰徐宁胡勇祁勇刚周梦舟吴茜
Owner HUBEI UNIV OF TECH
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