A method for biosynthesizing piperonal with recombinant escherichia coli

Abstract

The invention relates to a heliotropin biosynthesis method utilizing recombinant escherichia coli, and belongs to the technical filed of biological engineering. The method is characterized in that anoxidase gene derived from pseudomonas is introduced into escherichia coli. The recombinant bacterium can catalyze heliotropin synthesis from propenylbenzene, and the concentrated of the produced heliotropin is 9.15 g / L. The method in which the heliotropin is synthesized by utilizing recombinant oxidase has characteristics of a short production period, and simple extraction, and has a good industrial production prospect.

Description

technical field

[0001] The invention relates to a method for biosynthesizing piperonal with recombinant Escherichia coli, belonging to the technical field of bioengineering. Background technique

[0002] Piperonal, also known as jasmonal, has a chemical name of 3,4-methylenedioxybenzaldehyde and has a sweet cherry and vanilla aroma. Piperonal is widely used in food flavors, cosmetics industry, pharmaceutical industry, agriculture and electroplating industry. Natural jasmonal mainly exists in plants such as melon, black locust, fragrant pepper, vanilla bean, etc., but the content is very small, and people cannot directly extract it from plants.

[0003] The synthesis of jasmonal in the industry is mainly divided into two categories: one is the semi-synthetic method using safrole and piperine extracted from plants as raw materials; the other is the use of catechol, p- Total synthesis of methyl phenol raw materials. Among them, the method of synthesizing piperonal with sasro...

Examples

Embodiment Construction

[0017] The following is the enzyme activity assay of recombinant bacteria and piperonal assay method

[0018] Enzyme activity measurement method: Add appropriate amount of induced recombinant bacteria and safrole to the reaction system, react at 30°C for 1 hour, take a certain volume of reaction solution, add an equal volume of methanol to mix, stop the reaction, and centrifuge at 1000rpm for 5 minutes. The supernatant was filtered through a 0.22 μM organic membrane, and the content of piperonal was detected by HPLC.

[0019] Definition of enzyme activity: The defined amount of enzyme required to generate 1mmol / L piperonal per 1h is 1U, and the specific enzyme activity is the enzyme activity contained in 1g of stem cells.

[0020] HPLC detection conditions: Amethst C18-H reverse column (4.6mm×250mm, 5μm), with 60% acetonitrile and 0.1% formic acid as the mobile phase, the temperature of the column oven is 35°C, the injection volume is 10μL, 270nm Detection of piperonal conten...

technical field

[0001] The invention relates to a method for biosynthesizing piperonal with recombinant Escherichia coli, belonging to the technical field of bioengineering. Background technique

[0002] Piperonal, also known as jasmonal, has a chemical name of 3,4-methylenedioxybenzaldehyde and has a sweet cherry and vanilla aroma. Piperonal is widely used in food flavors, cosmetics industry, pharmaceutical industry, agriculture and electroplating industry. Natural jasmonal mainly exists in plants such as melon, black locust, fragrant pepper, vanilla bean, etc., but the content is very small, and people cannot directly extract it from plants.

[0003] The synthesis of jasmonal in the industry is mainly divided into two categories: one is the semi-synthetic method using safrole and piperine extracted from plants as raw materials; the other is the use of catechol, p- Total synthesis of methyl phenol raw materials. Among them, the method of synthesizing piperonal with sasro...

Examples

Embodiment Construction

[0017] The following is the enzyme activity assay of recombinant bacteria and piperonal assay method

[0018] Enzyme activity measurement method: Add appropriate amount of induced recombinant bacteria and safrole to the reaction system, react at 30°C for 1 hour, take a certain volume of reaction solution, add an equal volume of methanol to mix, stop the reaction, and centrifuge at 1000rpm for 5 minutes. The supernatant was filtered through a 0.22 μM organic membrane, and the content of piperonal was detected by HPLC.

[0019] Definition of enzyme activity: The defined amount of enzyme required to generate 1mmol / L piperonal per 1h is 1U, and the specific enzyme activity is the enzyme activity contained in 1g of stem cells.

[0020] HPLC detection conditions: Amethst C18-H reverse column (4.6mm×250mm, 5μm), with 60% acetonitrile and 0.1% formic acid as the mobile phase, the temperature of the column oven is 35°C, the injection volume is 10μL, 270nm Detection of piperonal conten...

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