Sphingomonas sp., extracellular product thereof, preparation method and application thereof

A sphingomonas, extracellular product technology, applied in the preparation of its extracellular products and extracellular products, extracellular products in skin cell proliferation, sphingomonas, repair and anti-aging aspects. In the field of application, it can solve the problem of low effect of active ingredients, and achieve the effect of delaying cell aging, repairing cell aging, and promoting the proliferation of epidermal cells.

Active Publication Date: 2019-01-18
JALA GROUP CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This extraction method limits the active ingredient to come from bacteria, the effect of the active ingredient is low, and the extraction process requires the use of organic reagents, which poses certain health risks to workers and consumers

Method used

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  • Sphingomonas sp., extracellular product thereof, preparation method and application thereof
  • Sphingomonas sp., extracellular product thereof, preparation method and application thereof
  • Sphingomonas sp., extracellular product thereof, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Screening and identification of Sphingomonas: After enrichment from Riduo hot spring water in the Himalayas, use tryptic soybean agar plate (OXOID, casein trypsin hydrolyzate 15.0g / L, soybean papain digest 5.0g / L, sodium chloride 5.0g / L, agar 15.0g / L) separation, screening for suspected single colonies, and then performing 2 times of purification of streaked strains; the purified strains were sent to Shanghai Meiji Biotechnology Co., Ltd. For identification of bacterial species, DNA was extracted using a bacterial genomic DNA kit (AxyPrep, bacterial genomic DNA miniprep kit AP-MN-BT-GDNA-250) as a template for PCR amplification. The primers used in PCR were 16S universal primer 27F (the sequence is 5'-AGA GTT TGA TCM TGG CTC AG-3') and 1492R (the sequence is 5'-TAC GGY TAC CTT GTT ACG ACT T-3').

[0030] PCR system:

[0031] Reagent

volume

10×Ex Taq buffer

2.0μl

2.5mM dNTP Mix

1.6μl

10mM 27F

0.8μl

10mM 1492R

0.8...

Embodiment 2

[0036] Extracellular product preparation: configuration medium (sucrose 20g / L, yeast extract 1g / L, K 2 HPO 4 2g / L,MgSO 4 ·7H 2(00.1g / L, regulate pH about 7.0) subpackage 50ml is in 250ml Erlenmeyer flask, get the sphingomonas (Sphingomonas sp.) CCTCC NO.M 2017211 JA-MS-001-A- that embodiment 1 screens out 04 strains in a constant temperature shaking incubator at 28-37°C (30°C is the best), about 220rpm for 18-24h, strain concentration OD 600 =0.6~0.8, inoculation ratio 1:100 (volume ratio), after fermentation, centrifuge at 5000rpm for 10~15min, remove the bacteria, collect the supernatant, which is the crude extract of extracellular products, pre-freeze overnight at -20°C, and vacuum freeze After drying overnight, the extracellular product powder was obtained, which was frozen at -20°C. When in use, sterile deionized water was configured to 10 mg / ml for subsequent experiments.

[0037] The medium used for the above-mentioned fermentation can also be a conventional mediu...

Embodiment 3

[0040] Keratinocyte culture: Human keratinocytes (Keratinocyte, KC) use complete medium (Thermo) at 37°C in 5% CO 2 Culture in a cell culture box, and inoculate after trypsinization when the plating rate reaches 90%.

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PUM

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Abstract

The invention provides a Sphingomonas sp., an extracellular product thereof, and a preparation method. The sphingomonas sp. Is Sphingomonas sp. JA. Deposited as CCTCC NO: M 2017211. MS-001-A-04 strain. The invention also discloses the application of the extracellular product of Sphingomonas sp. In skin cell proliferation, repair and anti-aging, as well as an anti-aging preparation containing the extracellular product of Sphingomonas sp. For promoting cell proliferation and/or migration.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a sphingomonas, its extracellular product and the preparation of the extracellular product, and the application of the extracellular product in skin cell proliferation, repair and aging delay. Background technique [0002] Sphingomonas was isolated and identified by scientists in 1990. Until 2001, it was divided into four genera in taxonomy due to its physical characteristics and phylogenetic characteristics: Sphingomonas, Sphingomonas, and Sphingomonas. Sphingobium, Novosphingobium, Sphigopyxis. Sphingomonas (Sphingomonas) is a Gram-negative bacterium, rod-shaped, typical aerobic, chemically heterotrophic, with a unique respiratory metabolic pathway-coenzyme Q10, and the cell membrane mainly contains glycosphingolipids instead of lipids Polysaccharides, which usually secrete pigments that give colonies a yellow color. Sphingomonas spp. has received increasing attention...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04A61Q19/08C12R1/01
CPCA61Q19/08A61K8/99C12P1/04C12R2001/01C12N1/205
Inventor 齐佩瑾吴越宋肖洁周春霞王宇史晓婷
Owner JALA GROUP CORPORATION
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