Method for quickly producing carotenoid by utilizing microaerobic fermentation of rhodobacter sphaeroides

A technology of photosynthetic bacteria and micro-aerobic fermentation, applied in the field of fermentation, can solve problems such as affecting efficiency and time-consuming, and achieve the effect of short fermentation time and shortening fermentation cycle

Active Publication Date: 2015-07-29
SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although photosynthetic bacteria are rich in natural carotenoids, photosynthetic bacteria have not been really used to produce natural carotenoids on an industrial scale
One of the important reasons is that the current method of photosynthetic bacteria fermentation to produce natural carotenoids is the light fermentation method, which is very time-consuming, and the fermentation period is 1-2 weeks, or even longer, which seriously affects the efficiency.

Method used

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  • Method for quickly producing carotenoid by utilizing microaerobic fermentation of rhodobacter sphaeroides
  • Method for quickly producing carotenoid by utilizing microaerobic fermentation of rhodobacter sphaeroides
  • Method for quickly producing carotenoid by utilizing microaerobic fermentation of rhodobacter sphaeroides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Activation of photosynthetic bacteria

[0034] The photosynthetic bacteria (Rb.sphaeroides) stored at -80℃ were streaked and cultured on MMS solid medium, and cultivated at 30℃ for about 4 days to grow a single red colony; then, the single red colony was picked and inoculated with 10-20mL MMS In a 50mL Erlenmeyer flask of liquid culture medium, culture for 18h at 30°C and 200rpm until the photosynthetic bacteria grow to pink.

Embodiment 2

[0035] Example 2. Rapid production of carotenoids by microaerobic fermentation

[0036] With 8% inoculation amount, the photosynthetic bacteria activated in Example 1 were respectively inoculated into two 250 mL conical flasks containing 100 mL MMS medium, and aerobic fermentation was carried out to OD600 at a temperature of 30°C, a rotation speed of 250 rpm, and dark conditions. =0.8, then combine the two bottles of fermentation broth into one bottle so that the volume of the fermentation broth accounts for 80% of the total volume of the Erlenmeyer flask, and then continue micro-aerobic fermentation for 36 hours at a temperature of 30°C, a rotation speed of 150 rpm, and darkness. , Quickly induce a large amount of synthetic carotenoids to ferment, the color of the fermentation broth changes to dark red, such as figure 1 Shown.

[0037] In this example, the conditions for aerobic fermentation are 200-250 rpm and temperature 30-32°C, and the end point OD600 of aerobic fermentation i...

Embodiment 3

[0038] Example 3 Extraction of carotenoids

[0039] The fermentation broth of Example 2 was centrifuged at 10000 rpm and 4°C for 10 min, the supernatant was discarded, rinsed with distilled water once, dried and weighed, and the results showed that the bacterial cell yield exceeded 6 g / L. Compared with the production of carotenoids by the light fermentation method, this method only uses 36 hours of fermentation time, which greatly shortens the fermentation time. At the same time, the carotenoid biosynthesis can be quickly judged by the color of the fermentation broth.

[0040] Then add an appropriate amount of extractant with a volume ratio of acetone to methanol of 7:2 to resuspend the bacteria, and ultrasonically break; the ultrasonic breaking conditions are: power 300W, hit 3s, interval 5s, ice bath, lasting 30min. After crushing, centrifuge at 10000rpm and 4℃ for 10min, collect the supernatant, make up the extractant with the volume ratio of acetone to methanol 7:2 according to...

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Abstract

The invention discloses a method for quickly producing carotenoid by utilizing microaerobic fermentation of rhodobacter sphaeroides. The method comprises the following particular steps that the activated rhodobacter sphaeroides is inoculated to an MMS (methylmercuric sulfate) culture medium; aerobic culture is performed until OD600 (optical density 600) is 0.6-0.8; a culture condition is adjusted to be microaerobic; the carotenoid is induced to be quickly and greatly synthesized; and then extraction and purification are performed. The time for producing the carotenoid by the method is short and only 36h; the yield of thallus is up to 6g/L; the yield and the productivity of the carotenoid are 8.287mg/L and 1911mug/g respectively; the carotenoid has good capacity for removing DPPH (1,1-diphenyl-2-picryl-hydrazyl) radicals; and IC50 (half maximal inhibitory concentration) is about 8mug/mL.

Description

Technical field [0001] The invention belongs to the field of fermentation, and specifically relates to a method for rapidly producing carotenoids by using photosynthetic bacteria micro-aerobic fermentation. Background technique [0002] Carotenoids are a type of fat-soluble pigments, most of which are insoluble in water and soluble in organic solvents. They are a large class of pigments in the global ecosystem and are one of the most widely distributed and important pigments in nature. Carotenoids can increase the body's immune function and the connection and communication between cells, and have important functions such as antioxidant function, anti-cancer function, and energy transmission. Because of its antioxidant properties, it has become a new type of antioxidant active substance that has been widely concerned in the fields of international drugs, dietary supplements, and functional foods, and is known as the "new nobility" in the antioxidant era in the 21st century. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P23/00C12R1/01
Inventor 赵志平聂鑫张利肖长江赵阳刘兴波
Owner SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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