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Method for extracting pyrroloquinoline quinone from fermentation liquor

A technology of pyrroloquinoline quinone and fermentation liquid, which is applied in the field of biological fermentation engineering, can solve problems such as unstable product content, increased production costs, and high maintenance costs, and achieves the effects of easy automatic operation, reduced usage, and reduced equipment maintenance costs

Inactive Publication Date: 2020-07-24
山东惠仕莱生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above-mentioned several extraction methods all used means such as the adsorption resin of different properties, gel resin, polyamide resin and magnetic adsorption separation in different procedures, and PQQ is purified, and the adsorption resin is in the actual operation process, due to the need to Collect and elute the active peak of PQQ, so the actual operation process has higher requirements for production technical operators, which may lead to unstable product content. At the same time, there will be obvious dead adsorption in the resin after use, and it must be used after each use. Concentration of organic solvent to regenerate, in order to ensure the next separation performance, high maintenance costs, increase production costs

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 5L fermentation broth, PQQ content is 10g, centrifuge to collect 4.91L of filtrate, adjust the pH value to 3 with 10% hydrochloric acid, add 50g of diatomaceous earth, stir at 15°C for 1.5h, filter and collect 5L of filtrate, and adjust with 10% NaOH solution pH value is 8, add 250g of activated carbon, stir at 15°C for 2 hours, filter with ceramic membrane, the pore size of the ceramic membrane is 30-40nm, collect 8L of filtrate, and treat the filtrate with an ultrafiltration membrane with a cut-off flow of 1000Da to obtain 16L of filtrate, continue to use molecular weight cut-off Treat the filtrate with a 200Da nanofiltration membrane, concentrate until the PQQ content is 6g / L, collect 1.5L of the concentrated solution, continue to concentrate under reduced pressure, control the material temperature at 20°C, concentrate until the PQQ content is 20g / L, and use 10% dilute hydrochloric acid Adjust the pH value to 3.5, and crystallize at 2°C for 36 hours to obtain a PQQ pr...

Embodiment 2

[0030] 5L fermentation broth, PQQ content is 10g, centrifuge to collect 4.93L of filtrate, adjust the pH value to 3.5 with 10% hydrochloric acid, add 100g of diatomaceous earth, stir at 15°C for 1.5h, filter and collect 5L of filtrate, and then adjust with 10% NaOH solution The pH value is 7.5, add 150g of activated carbon, stir at 15°C for 1 hour, filter with ceramic membrane, the pore size of the ceramic membrane is 30-40nm, collect 8L of filtrate, and treat the filtrate with an ultrafiltration membrane with a cut-off flow of 800Da to obtain 16L of filtrate, continue to use molecular weight cut-off Treat the filtrate with a 300Da nanofiltration membrane, concentrate until the PQQ content is 5g / L, collect 2L of the concentrate, continue to concentrate under reduced pressure, control the temperature of the material at 20°C, concentrate until the PQQ content is 10g / L, and adjust with 10% dilute hydrochloric acid The pH value was 3, crystallized at 4°C for 24 hours, and the purit...

Embodiment 3

[0032] 5L fermentation broth, PQQ content is 10g, centrifuge to collect 4.8L of filtrate, adjust the pH value to 3.3 with 10% hydrochloric acid, add 80g of diatomaceous earth, stir at 15°C for 1.5h, filter and collect 5L of filtrate, and then adjust with 10% NaOH solution The pH value is 7.5, add 200g of activated carbon, stir at 15°C for 1.5h, filter with a ceramic membrane, the pore size of the ceramic membrane is 30-40nm, collect 8L of filtrate, and treat the filtrate with an ultrafiltration membrane with a cut-off flow of 900Da to obtain 16L of filtrate, continue to use cut-off Treat the filtrate with a molecular weight of 250Da nanofiltration membrane, concentrate until the PQQ content is 5.5g / L, collect 1.8L of the concentrated solution, continue to concentrate under reduced pressure, control the temperature of the material at 25°C, concentrate until the PQQ content is 15g / L, and use 10% Adjust the pH value to 3 with dilute hydrochloric acid, and crystallize at 4°C for 48...

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Abstract

A method for extracting pyrroloquinoline quinone from fermentation broth mainly comprises the following steps: pretreating the fermentation broth to obtain a filtrate, adjusting the pH value of the filtrate to 2.5-3.5 with an acid, adding an adsorbent, and carrying out first filtration to obtain a filtrate; adjusting the pH value of the filtrate to 7.5-8, adding an adsorbent, and performing filtering for the second time to obtain a filtrate; and carrying out ultrafiltration and nanofiltration on the filtrate to obtain a nanofiltration concentrated solution, carrying out reduced pressure concentration on the nanofiltration concentrated solution, adjusting the pH value to 3-3.5, and carrying out low-temperature crystallization to obtain the product. The purity of the final product PQQ is greater than or equal to 98%, the total yield is greater than or equal to 85%, and multistage membrane equipment is adopted in the extraction process, so that the extraction operation is simpler, the automatic operation is easier, the equipment maintenance cost is reduced, the use amount of organic solvents in production is reduced, and the production cost of enterprises is reduced.

Description

technical field [0001] The invention relates to the technical field of biological fermentation engineering, in particular to a method for extracting pyrroloquinoline quinone from fermentation broth. Background technique [0002] Pyrroloquinoline quinone (Pyrroloquinoline quinone, PQQ) is a water-soluble near-position quinone compound, which is the third redox compound discovered after pyridine nucleotides (NAD, NADP) and riboflavin (FMN, FAD). Enzyme coenzyme. PQQ widely exists in living organisms, has the functions of stimulating the growth of nerve factors, scavenging free radicals, etc., and has important development prospects in the fields of food and medical health products. In the early days, PQQ was mainly produced by chemical synthesis, but the synthesis method has disadvantages such as complicated steps, high cost, and large pollution, and is gradually being replaced by microbial fermentation. [0003] There are many bacterial species that can ferment and produce ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D471/04
CPCC07D471/04
Inventor 孙荣刘建民任明张雷
Owner 山东惠仕莱生物科技有限公司
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