Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for selectively separating cinnamic acid in phenylalanine bioconversion broth and cyclically utilizing conversion broth

A technology for biotransformation and phenylalanine, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as low biotransformation rate, reduce substrate inhibition, reduce extraction cost, process simple effect

Active Publication Date: 2016-01-06
NANJING FORESTRY UNIV
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are not many studies on the preparation of biological methods. One of the factors is that cinnamic acid has an obvious inhibitory effect on phenylalanine ammonia lyase, and the low biotransformation rate leads to a higher concentration of phenylalanine and With a lower concentration of cinnamic acid, finding a method to quickly and effectively separate cinnamic acid from phenylalanine solution is not only beneficial to relieve the product inhibition phenomenon in the reaction, but also has great significance for the industrialization of biological preparation of cinnamic acid

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
  • Method for selectively separating cinnamic acid in phenylalanine bioconversion broth and cyclically utilizing conversion broth
  • Method for selectively separating cinnamic acid in phenylalanine bioconversion broth and cyclically utilizing conversion broth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] 1. Weigh 0.6g of phenylalanine, put it into a 50ml Erlenmeyer flask, add 30ml of Tris buffer, adjust the pH to 9.0 with hydrochloric acid, and add 0.26g of recombinant Escherichia coli containing phenylalanine ammonia-lyase [ References: ZangY., JiangT., CongY., ZhengZJ., OuyangJ*. Molecular Characterization of a Recombinant Zeamays Phenylalanine Ammonia-Lyase (ZmPAL2) and Its Application in trans-Cinnamic Acid Production from L-Phenylalanine. Appl Biochem Biotechnol., 2015, 196:924 , shake well, react at a temperature of 55° C. for 2 h, and rotate at 200 rpm.

[0030] 2. Obtain a whole-cell transformation solution with an L-phenylalanine content of 5.69 g / L and a cinnamic acid content of 14.94 g / L, centrifuge at 8000 rpm for 5 minutes for solid-liquid separation, and mark the supernatant as supernatant I, solid-phase bacteria The body was marked as Sludge II and removed from the reaction system for the secondary reaction.

[0031] 3. Add analytically pure hydrochloric...

Embodiment 2

[0036] 1. Weigh 1.2g of phenylalanine, put it into a 50ml Erlenmeyer flask, add 30ml of Tris buffer, adjust the pH to 9.0 with hydrochloric acid, and add 0.26g of recombinant Escherichia coli containing phenylalanine ammonia-lyase, The final concentration OD is 20, shake well, and react at a temperature of 55° C. for 2 hours with a rotation speed of 200 rpm.

[0037] 2. Obtain the whole-cell transformation solution with the L-phenylalanine content of 23.50g / L and the cinnamic acid content of 13.19g / L, centrifuge at 8000rpm for 5min for solid-liquid separation, and mark the supernatant as supernatant I, solid-phase bacteria The body is marked as Sludge II, removed from the reaction system and used for the secondary reaction.

[0038]3. Add analytically pure hydrochloric acid (36%) to the supernatant liquid I at room temperature to adjust the pH to 4.5, oscillate and stir for 5 minutes, and rotate at 150 rpm.

[0039] 4. Centrifuge the solution I with white precipitate at 8000r...

Embodiment 5

[0054] Embodiment 5: the infrared chromatogram of cinnamic acid finished product

[0055] The sample that embodiment 2 obtains absolutely dry carries out infrared chromatographic analysis, and the results are shown in figure 1 shown.

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

No PUM Login to View More

Abstract

The invention discloses a method for selectively utilizing cinnamic acid in phenylalanine bioconversion broth and cyclically utilizing conversion broth. The method comprises the following steps: performing solid-liquid separation on a phenylalanine bioconversion broth to respectively obtain a clear liquid I and cell thallus; regulating the pH value of the clear liquid I to 4.5 with acid, and stirring the clear liquid I at room temperature until white precipitate is separated; performing solid-liquid separation on the mixed system to obtain a clear liquid II and precipitation, and performing washing and freeze-drying on the precipitation to obtain a cinnamic acid sample; and regulating the pH value of the clear liquid II to the value required by bioconversion with alkali, mixing the clear liquid II with the cell thallus, and continuously performing bioconversion to prepare the cinnamic acid. By adopting the method, the method can be used for reducing the inhibition effect of a primer and cyclically utilizing the conversion broth and cells, the yield is up to 95 percent, and the product purity is up to over 98 percent. The method is simple and easy in process, and can be used for greatly reducing the extraction cost and realizing multiple times of cyclic utilization of resting cells and conversion broth.

Description

technical field [0001] The invention belongs to the technical field of bioseparation, and relates to a method for separating products and reusing the transformation solution in the process of preparing cinnamic acid through biotransformation. Background technique [0002] Cinnamic acid, also known as β-phenylacrylic acid, 3-phenyl-2-acrylic acid. It is an organic acid isolated from cinnamon bark or benzoin. Phenylacrylic acid produced by the deamination degradation of phenylalanine in plants. [0003] The main uses of cinnamic acid: a. Application in flavors and fragrances: cinnamic acid itself is a kind of spice, which has a good aroma retention effect. It is usually used as a fragrance ingredient to make the aroma of the main spice more volatile. Essence, cherry essence, fruit essence, and flower essence are blended and used; it can be used as an aromatic mixture in soap, shampoo, washing powder, and daily cosmetics; b. The role in medicine: cinnamic acid can be used to ...

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
IPC IPC(8): C12P7/40C12R1/19
Inventor 欧阳嘉臧颖郑兆娟李鑫朱均均徐勇勇强
Owner NANJING FORESTRY UNIV