Bio-enzyme synthesis method and application of cosmetic-grade hexose-6-phosphoric acid composition

A cosmetic-grade, bio-enzymatic technology, applied in the field of biological composition preparation, can solve problems such as different biochemical metabolism pathways and differences in anti-aging functional components, and achieve the effects of reducing process difficulty, saving production costs, and reducing process routes

Inactive Publication Date: 2022-07-01
北京焉支山科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biochemical metabolism pathways of the two are different, which makes the

Method used

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  • Bio-enzyme synthesis method and application of cosmetic-grade hexose-6-phosphoric acid composition
  • Bio-enzyme synthesis method and application of cosmetic-grade hexose-6-phosphoric acid composition
  • Bio-enzyme synthesis method and application of cosmetic-grade hexose-6-phosphoric acid composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1: Genetically engineered bacteria I prepare hexokinase crude enzyme liquid by fermentation

[0067] (1) Bacterial fermentation:

[0068] The genetically engineered bacteria I capable of heterologously expressing polyphosphate-dependent mannokinase were inoculated into 50 mL of LB liquid medium (50 mg / L Kana), fermented for 8 h, at a temperature of 37 °C, and at a stirring speed of 180 rpm; The fermentation product was inoculated into 2L of LB liquid medium (50mg / L Kana) according to 1‰, and the air flow was controlled to be 4Nm under the conditions of temperature 30℃ and pH 7 3 / h; after the dissolved oxygen in the fermentation system is stabilized at 10%, an inducer IPTG is added, and the above fermentation conditions are maintained for 12 hours to obtain a fermentation broth.

[0069] (2) Preparation of crude enzyme solution:

[0070] The fermentation broth was centrifuged at 4°C and a rotating speed of 8000rpm to collect the cells, and the cells were re...

Embodiment 2

[0076] Example 2: Preparation of phosphate mannose isomerase crude enzyme liquid by fermentation of genetically engineered bacteria II

[0077] Bacterial fermentation:

[0078] The genetically engineered bacteria II capable of heterologously expressing phosphomannose isomerase was inoculated into 50 mL of LB liquid medium (50 mg / L Kana), and fermented for 8 h at a temperature of 37°C and a stirring speed of 180 rpm; 0.5‰ was inoculated into 2L of LB liquid medium (50mg / L Kana), and the air flow was controlled to be 4Nm under the conditions of temperature of 30℃ and pH of 7 3 / h; after the dissolved oxygen in the fermentation system is stabilized at 10%, an inducer IPTG is added, and the above fermentation conditions are maintained for 12 hours to obtain a fermentation broth.

[0079] Preparation of crude enzyme solution:

[0080] Carry out according to the method described in Example 1.

[0081] Enzyme activity assay:

[0082] Accurately weigh 1mL of the crude enzyme solut...

Embodiment 3

[0086] Embodiment three: the joint fermentation preparation of mixed enzyme crude enzyme liquid

[0087] (1) Bacterial fermentation:

[0088]The genetically engineered bacteria I capable of heterologously expressing polyphosphate-dependent mannokinase was inoculated into 50 mL of LB liquid culture medium (50 mg / L Kana); at the same time, the heterologous expression of mannose phosphate isomer The genetically engineered bacteria II of the enzyme was inoculated into 50 mL of LB liquid medium (50 mg / LKana); the fermentation conditions of both were 37 ° C, 8 h, and 180 rpm; and then the fermentation product of the genetically engineered bacteria I was 1‰, genetically engineered The fermentation product of Bacteria II was inoculated into 2L of LB liquid medium (50mg / L Kana) according to 0.5‰, and the air flow was controlled to be 4Nm under the conditions of temperature of 30℃ and pH of 7. 3 / h; after the dissolved oxygen in the fermentation system is stabilized at 10%, an inducer ...

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Abstract

The embodiment of the invention discloses a biological enzyme synthesis method and application of a cosmetic-grade hexose-6-phosphoric acid composition, and the method comprises the following steps: transforming a coding gene 17 of polyphosphate dependent kinase into escherichia coli to construct a genetically engineered bacterium I; the method comprises the following steps: transforming a coding gene BSMPI of phosphomannose isomerase into bacillus subtilis, and constructing a genetically engineered bacterium II; carrying out combined fermentation on the genetically engineered bacterium I and the genetically engineered bacterium II to prepare a crude enzyme solution of the compound enzyme; and carrying out enzyme catalytic reaction on the crude enzyme liquid of the compound enzyme to prepare the cosmetic-grade hexose-6-phosphoric acid composition. According to the method, biological enzyme does not need to be purified, and an enzyme catalysis product does not need to be refined and dried, so that the production and application cost is greatly reduced; in addition, cell experiments, skin tests and other means prove that the hexose-6-phosphoric acid composition disclosed by the invention has good effects of improving cell energy metabolism efficiency, promoting skin repair and regeneration, relieving skin wrinkles and the like.

Description

technical field [0001] The invention relates to the technical field of biological composition preparation, in particular to a biological enzymatic synthesis method and application of a cosmetic grade hexose-6-phosphate composition. Background technique [0002] Delaying skin aging is one of the claims of many cosmetic products. Skin aging is generally considered to be the result of a combination of factors including exogenous environmental stimuli and endogenous gene attenuation. At present, there are free radical theory, aging gene theory and mitochondrial DNA damage theory in this field to theoretically explain and physiologically verify the phenomenon of skin aging. No matter which theory or theory, its research foundation is based on the metabolism of skin surface cells, and the material and energy metabolism process involving mitochondria is one of the focuses of research. [0003] On the other hand, hexose phosphate is one of the important substances for mitochondria...

Claims

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Application Information

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IPC IPC(8): C12P19/02C12P19/24C12P39/00C12N15/75C12N15/70C12N15/54C12N15/61A61Q19/08A61K8/64A61K8/60A61K8/34A61K8/24C12R1/125C12R1/19
CPCC12P19/02C12P19/24C12P39/00C12N15/70C12N15/75C12N9/1205C12N9/90A61K8/60A61K8/24A61K8/64A61K8/345A61Q19/08C12Y207/01007C12Y503/01008A61K2800/5922
Inventor 殷昊
Owner 北京焉支山科技有限公司
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