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Sphingomonas and method for producing carotenoids therewith

A technology of sphingomonas, sphingobiumsp.kib, which is applied in the preparation of carotenoids and in the field of biology, and can solve the problems of long cycle time, low animal utilization rate, and large floor area

Inactive Publication Date: 2020-08-11
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The animal utilization rate of chemical synthesis is low; the waste extraction cost of crustaceans is high, and the yield is low; the growth conditions of Haematococcus pluvialis are harsh, the cycle is long, and the land occupation is large

Method used

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  • Sphingomonas and method for producing carotenoids therewith
  • Sphingomonas and method for producing carotenoids therewith
  • Sphingomonas and method for producing carotenoids therewith

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Breeding of Sphingomonas phytoene-producing strain SP-Phy.

[0082] The starting strain KIB of Sphingomonas: isolated by our laboratory, is a pollutant on the plate medium when the marine microalgae Aurantiochytrium sp.SK4 is cultivated. The plate medium formula is: 5g glucose, 2g yeast extract, 50% artificial seawater, 8g agar powder and distilled water are fully mixed and the volume is adjusted to 1L with distilled water; sterilized at 121 degrees Celsius for 20 minutes; cultured in artificial seawater: 30gNaCl, 0.7gKCl, 10.8gMgCl 2 ·6H 2 O, 2.638gMgSO4, 0.756gCaCl 2 Mix well with distilled water and make up to 1L with distilled water. The bacterium is bright in color, and the colony is smooth and round. Pick a single clone in LB culture medium, cultivate at 28 degrees Celsius and 220 rpm. Sphingobium sp. was preserved on April 25, 2016 in the General Microbiology Center of China Committee for Culture Collection of Microorganisms (Address: Institute of Microbiology...

Embodiment 2

[0102] Breeding of Sphingomonas lycopene-producing strain SP-Lyc.

[0103] (1) The starting strain of Sphingomonas was inoculated into the slant of LB agar medium, and cultured at 28° C. for 48 hours. LB agar medium is peptone 10g / L, yeast extract 5g / L, sodium chloride 10g / L, pH 6.5;

[0104] (2) Pick a ring of bacteria from the activated slant and inoculate it into the shake flask, culture at 28°C, 220rpm for 12-18h, so that OD600=0.7-0.9, wherein the shake flask medium is TB medium: 24g of yeast extract / L, peptone 12g / L, glucose 4g / L, K 2 HPO 4 ·3H 2 O 16.416g / L,KH 2 PO 4 2.312g / L, pH 6.5;

[0105] (3) Take 10 mL of the above-mentioned cultured bacterial solution, collect the bacterial cells by centrifugation, wash the bacterial cells twice with an equal volume of phosphate buffered saline solution, and collect the washed bacterial liquid by centrifugation;

[0106] (4) NTG mutagenesis: add 10 mL of phosphate-buffered saline to the cells obtained by centrifugation, ...

Embodiment 3

[0122] Breeding of zeaxanthin-producing strain SP-Zea from Sphingomonas sp.

[0123] (1) Inoculate the sphingomonas strain KIB into the slant of LB agar medium, and culture it at 28°C for 48 hours. The LB agar medium is 10g / L of peptone, 5g / L of yeast extract, and 10g / L of sodium chloride. L, pH 6.5;

[0124] (2) Pick a ring of bacteria from the activated slant and inoculate it into a shake flask, culture at 28° C., 220 rpm for 12-18 hours to make OD600 = 0.7-0.9, wherein the shake flask medium is TB. TB medium is yeast extract 24g / L, peptone 12g / L, glucose 4g / L, K 2 HPO 4 ·3H 2 O 16.416g / L, KH 2 PO 4 2.312g / L, pH 6.5;

[0125] (3) Take 10 mL of the above-mentioned cultured bacterial solution, collect the bacterial cells by centrifugation, wash the bacterial cells twice with an equal volume of phosphate buffered saline solution, and collect the washed bacterial liquid by centrifugation;

[0126] (4) NTG mutagenesis: add 10 mL of phosphate-buffered saline to the cells o...

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Abstract

The invention provides a sphingomonas with a preservation number of CGMCC No.12394; a method for cultivating a carotenoid-producing strain, its application in the production of carotenoids, and its use in the industrial production of astaxanthin in the application. In the sphingomonas producing nodoc xanthin of the present invention, the content of nodoc xanthin can reach 3.98±0.5 mg / g, accounting for 90% or more of the total pigment. The colorless phytoene-producing bacteria SP-Phy, the red lycopene-producing bacteria SP-Lyc, and the yellow zeaxanthin-producing bacteria SP-Zea selected and bred by the present invention are successively passed on for more than 5 generations respectively. The yields of lycopene, lycopene and zeaxanthin are stable, and the content can reach 3.7±0.5mg / g, 3.9±0.5mg / g, 4.0±0.5mg / g respectively, accounting for 90% and above of the total pigment; The produced zeaxanthin-producing strain SP-Zea introduced the ketolase gene and transformed it into an astaxanthin-producing strain SPZ-AST, with an astaxanthin content of 4.1±0.5 mg / g, accounting for 90% or more of the total pigment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the field of carotenoid preparation, more specifically, to a nodoc xanthin-producing bacterium Sphingomonas and a method for producing carotenoids with it. Background technique [0002] Carotenoids are highly unsaturated polyenes containing a series of conjugated double bonds and methyl branches. The color of a pigment varies with the number of conjugated double bonds. Due to their unique chemical structures, carotenoids play an important role in healthcare. Carotenoids have anti-oxidation effects, can scavenge singlet oxygen and peroxide free radicals, and reduce free radical damage to nucleic acids, proteins, and lipids, thereby delaying aging and preventing diseases such as thrombosis and arteriosclerosis. Carotenoids can enhance immune function, increase the activity of T lymphocytes, and assist B lymphocytes to produce antibodies. Many medical studies have shown that caroteno...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N15/01C12N1/21C12P23/00C12R1/01
CPCC12N9/0077C12N15/01C12P23/00C12Y114/15802C12N1/205C12R2001/01
Inventor 黄俊潮刘萌萌叶景润赵启超
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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