Production method for alpha-arbutin with high optical purity and glucan with high adhesion

Abstract

The invention discloses a co-production method for alpha-arbutin with high optical purity and glucan with high adhesion by using leuconostoc pseudomesenteroides. The method takes sucrose and hydroquinone as substrates to produce the alpha-arbutin and the glucan by fermenting in one step under the weak acid environment, and comprises the following steps: 1) first level seed culture: inoculating a leuconostoc pseudomesenteroides solution in a seed medium with 0.1 to 0.2% inoculation amount, and cultivating at 37 DEG C and 250rpm for 10 to 12 hours; 2) second level seed culture: transmitting a first level seed solution to the seed medium with 1 to 2% inoculation amount, and cultivating at 37 DEG C and 250rpm for 10 to 12 hours; 3) fermentation culture: inoculating a second level seed solution in a fermentation medium with 5 to 8% inoculation amount, adjusting pH in a fermentation process to be 6.0 to 6.8, blowing air in the whole fermentation process, controlling the air flow to be 0.1 to 0. 3vvm, and cultivating at 37 DEG C and 250 to 300rpm for 12 to 16 hours. According to the method, the alpha-arbutin with high optical purity and the glucan with high adhesion can be directly obtained by the fermentation of bacterial strain, and the method is high in substrate use rate, simple and quick in reaction, mild in condition, low in energy consumption, and suitable for industrial production.

Description

technical field

[0001] The invention relates to the technical field of biological fermentation, in particular to a method for producing alpha-arbutin with high optical purity and high-adhesive glucan by a strain of Leuconostoc mesenteroides. Background technique

[0002] There are two isomers of arbutin: α-arbutin and β-arbutin, both of which are epimers, chemically named 4-hydroxyphenyl-α-D-glucopyranoside and 4 -Hydroxyphenyl-β-D-glucopyranoside, the direction of the oxyglycoside bonds in space is opposite. Studies have shown that the strength and safety of α-arbutin in inhibiting tyrosinase is far greater than that of β-arbutin, and the whitening effect is 9 to 10 times that of the latter. Therefore, α-arbutin, as a high-efficiency cosmetic whitening agent, has been gradually used by major brands in the world: in 2002, Peutaharm launched a new active skin whitening agent containing α-arbutin, Japan Shiseido, DHC, etc. The brand has also launched a series of cosmetics co...

Examples

Embodiment 1

[0026] Leuconostocpseudomesenteroides (Leuconostocpseudomesenteroides) G123 was fermented in a 3L fermenter with a liquid volume of 2L, the fermentation condition was an initial pH of 6.5, the pH was controlled at 6.0 to 6.8 throughout the fermentation process, and 15g / L hydroquinone was sterilized separately and supplemented 10 times , 90g / L sucrose is sterilized separately and supplemented with hydroquinone at the same time, divided into 10 times, specifically including the following steps:

[0027] (1) Seed liquid culture:

[0028] First-level seed culture: inoculate the Leuconostoc pseudoenteritis bacteria liquid in the glycerol storage tube with an inoculum amount of 0.1-0.2% into the seed medium, and incubate at 37°C and 250rpm for 11 hours;

[0029] Secondary seed culture: transfer from the primary seed solution to the seed medium with an inoculation amount of 1-2%, and cultivate for 11 hours at 37°C and 250rpm;

[0030] The seed medium is: peptone 5.0g / L, yeast powder...

Embodiment 2

[0039] Leuconostoc pseudoenteritidis G123 was fermented in a 3L fermenter with a liquid volume of 2L. The fermentation condition was an initial pH of 6.5, and the pH was controlled at 6.0 to 6.8 throughout the fermentation process. 30g / L hydroquinone was sterilized separately and added in 10 times, 150g / L L sucrose is sterilized separately and supplemented with hydroquinone at the same time, divided into 10 times, specifically including the following steps:

[0040] (1) Seed liquid culture:

[0041] First-level seed culture: inoculate the Leuconostoc pseudoenteritis bacteria liquid in the glycerol storage tube with an inoculum amount of 0.1-0.2% into the seed medium, and incubate at 37°C and 250rpm for 10 hours;

[0042] Secondary seed culture: transfer from the primary seed solution to the seed medium with an inoculation amount of 1-2%, and cultivate for 10 hours at 37°C and 250rpm;

[0043] The seed medium is: peptone 4.0g / L, yeast powder 4.0g / L, sodium acetate 4.0g / L, Twee...

Embodiment 3

[0048] Leuconostoc pseudoenteritidis G123 was fermented in a 3L fermenter with a liquid volume of 2L. The fermentation condition was an initial pH of 6.5. The pH was controlled at 6.0 to 6.8 throughout the fermentation process. 10g / L hydroquinone was sterilized separately and added in 10 times, 80g / L L sucrose is sterilized separately and supplemented with hydroquinone at the same time, divided into 10 times, specifically including the following steps:

[0049] (1) Seed liquid culture:

[0050] First-level seed culture: inoculate the Leuconostoc pseudoenteritis bacteria solution in the glycerol storage tube with an inoculation amount of 0.1-0.2% into the seed medium, and cultivate at 37°C and 250rpm for 12 hours;

[0051] Secondary seed culture: transfer from the primary seed solution to the seed medium with an inoculation amount of 1-2%, and cultivate for 12 hours at 37°C and 250rpm;

[0052]The seed medium is: peptone 8.0g / L, yeast powder 8.0g / L, sodium acetate 8.0g / L, Twee...

technical field

[0001] The invention relates to the technical field of biological fermentation, in particular to a method for producing alpha-arbutin with high optical purity and high-adhesive glucan by a strain of Leuconostoc mesenteroides. Background technique

[0002] There are two isomers of arbutin: α-arbutin and β-arbutin, both of which are epimers, chemically named 4-hydroxyphenyl-α-D-glucopyranoside and 4 -Hydroxyphenyl-β-D-glucopyranoside, the direction of the oxyglycoside bonds in space is opposite. Studies have shown that the strength and safety of α-arbutin in inhibiting tyrosinase is far greater than that of β-arbutin, and the whitening effect is 9 to 10 times that of the latter. Therefore, α-arbutin, as a high-efficiency cosmetic whitening agent, has been gradually used by major brands in the world: in 2002, Peutaharm launched a new active skin whitening agent containing α-arbutin, Japan Shiseido, DHC, etc. The brand has also launched a series of cosmetics co...

Examples

Embodiment 1

[0026] Leuconostocpseudomesenteroides (Leuconostocpseudomesenteroides) G123 was fermented in a 3L fermenter with a liquid volume of 2L, the fermentation condition was an initial pH of 6.5, the pH was controlled at 6.0 to 6.8 throughout the fermentation process, and 15g / L hydroquinone was sterilized separately and supplemented 10 times , 90g / L sucrose is sterilized separately and supplemented with hydroquinone at the same time, divided into 10 times, specifically including the following steps:

[0027] (1) Seed liquid culture:

[0028] First-level seed culture: inoculate the Leuconostoc pseudoenteritis bacteria liquid in the glycerol storage tube with an inoculum amount of 0.1-0.2% into the seed medium, and incubate at 37°C and 250rpm for 11 hours;

[0029] Secondary seed culture: transfer from the primary seed solution to the seed medium with an inoculation amount of 1-2%, and cultivate for 11 hours at 37°C and 250rpm;

[0030] The seed medium is: peptone 5.0g / L, yeast powder...

Embodiment 2

[0039] Leuconostoc pseudoenteritidis G123 was fermented in a 3L fermenter with a liquid volume of 2L. The fermentation condition was an initial pH of 6.5, and the pH was controlled at 6.0 to 6.8 throughout the fermentation process. 30g / L hydroquinone was sterilized separately and added in 10 times, 150g / L L sucrose is sterilized separately and supplemented with hydroquinone at the same time, divided into 10 times, specifically including the following steps:

[0040] (1) Seed liquid culture:

[0041] First-level seed culture: inoculate the Leuconostoc pseudoenteritis bacteria liquid in the glycerol storage tube with an inoculum amount of 0.1-0.2% into the seed medium, and incubate at 37°C and 250rpm for 10 hours;

[0042] Secondary seed culture: transfer from the primary seed solution to the seed medium with an inoculation amount of 1-2%, and cultivate for 10 hours at 37°C and 250rpm;

[0043] The seed medium is: peptone 4.0g / L, yeast powder 4.0g / L, sodium acetate 4.0g / L, Twee...

Embodiment 3

[0048] Leuconostoc pseudoenteritidis G123 was fermented in a 3L fermenter with a liquid volume of 2L. The fermentation condition was an initial pH of 6.5. The pH was controlled at 6.0 to 6.8 throughout the fermentation process. 10g / L hydroquinone was sterilized separately and added in 10 times, 80g / L L sucrose is sterilized separately and supplemented with hydroquinone at the same time, divided into 10 times, specifically including the following steps:

[0049] (1) Seed liquid culture:

[0050] First-level seed culture: inoculate the Leuconostoc pseudoenteritis bacteria solution in the glycerol storage tube with an inoculation amount of 0.1-0.2% into the seed medium, and cultivate at 37°C and 250rpm for 12 hours;

[0051] Secondary seed culture: transfer from the primary seed solution to the seed medium with an inoculation amount of 1-2%, and cultivate for 12 hours at 37°C and 250rpm;

[0052]The seed medium is: peptone 8.0g / L, yeast powder 8.0g / L, sodium acetate 8.0g / L, Twee...

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