Method for preparing vanillyl alcohol by utilizing bacterial reduced vanillin

Abstract

The invention relates to a method for preparing vanillyl alcohol by utilizing bacterial reduced vanillin. The method comprises the following steps: (1) extracting a loop of bacillus aceticus from an agar slant culture medium for storing a strain, inoculating the bacillus aceticus to a liquid seed culture medium, and performing culturing for 12 to 24 hours at 20 to 30 DEG C to activate the strain; (2) inoculating 6 to 10 volume percent of seed solution into a liquid fermentation culture medium, and performing shaking table culturing or standing culturing for 12 to 48 hours at 20 to 30 DEG C; (3) when a great amount of flocculate is generated in the liquid fermentation culture medium, adding a filtered and disinfected vanillin solution into the culture medium until the final concentration of vanillin in the culture medium reaches 0.1 to 3.0 mmol / L, and performing shaking table culturing or standing culturing for 0.5 to 7 days at 20 to 35 DEG C to obtain the vanillyl alcohol. According to the method, the conversion rate is high, and can reach 50 to 98 percent, reaction conditions are mild and environment-friendly, and the method has a broad application prospect.

Description

technical field

[0001] The invention belongs to the field of preparation of vanillyl alcohol, in particular to a method for preparing vanillyl alcohol by reducing vanillin with bacteria. Background technique

[0002] Vanillyl alcohol, also known as vanillyl alcohol, vanillic alcohol, 4-hydroxy-3-methoxybenzyl alcohol, has a molecular weight of 154.16 and has a creamy sweet and coconut aroma. Compared with vanillin, the aroma is softer, longer-lasting, and more stable in nature. Vanillic alcohol is a recognized fragrance in the world and is suitable for almost all fragrance types. It can be widely used in food, tobacco and wine, beverages and daily chemical products, with unique and outstanding sweetness and aroma.

[0003] It generally exists in the tubers of Gastrodia elata Bl., an orchid plant, and is the active ingredient in Gastrodia elata. In terms of drug efficacy, it has obvious sedative effect, can reduce the spontaneous activity of mice, can prolong the sleep tim...

Examples

Embodiment 1

[0018] (1) Activated bacteria

[0019] Take a ring of Acetobacter xylinum ATCC23770 from the agar slant medium where the strains are preserved, and insert it into the liquid seed medium (mannitol, glucose, sucrose or glycerin 25g, peptone 3g, yeast extract 5g, water 1L, pH3 .5, sterilized at 121°C for 20 minutes), cultured at 160 rpm at 30°C for 12 hours to activate the bacteria;

[0020] (2) Expansion of bacterial strain cells

[0021] Add the seed liquid containing 6% (v / v) into the liquid medium (glucose, mannitol, sucrose or glycerin 25g, peptone 3g, yeast extract 5g, water 1L, pH3.5, sterilized at 121°C for 20min) medium, cultured on a shaking table at 20°C and 100r / min or statically cultured for 12 hours before use;

[0022] (3) Adding vanillin to prepare vanillyl alcohol

[0023] After a large amount of flocs are formed in the culture medium, add the sterile filtered vanillin solution to the culture medium respectively, so that the final concentration of vanillin in ...

Embodiment 2

[0027] (1) Activated bacteria

[0028] Take a ring of Acetobacter xylinum ATCC23770 from the agar slant medium where the strains are preserved, and insert it into the liquid seed medium (mannitol, glucose, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH 5.5, sterilized at 121°C for 20 minutes), cultured at 160 rpm at 30°C for 18 hours to activate the strains;

[0029] (2) Expansion of bacterial strain cells

[0030] Add the seed liquid containing 8% (v / v) into the liquid medium (glucose, mannitol, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH5.5, sterilized at 121°C for 20min ) in a shaker culture at 30°C and 250r / min or static culture for 18 hours before use;

[0031] (3) Adding vanillin to prepare vanillyl alcohol

[0032] After a large amount of flocs are formed in the culture medium, add the sterile filtered vanillin solution to the culture medium respectively, so that the final concentration of vanillin in the culture medium ...

Embodiment 3

[0034] (1) Activated bacteria

[0035] Take a ring of Acetobacter xylinum ATCC23770 from the agar slant medium where the strains are preserved, and insert it into the liquid seed medium (mannitol, glucose, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH 5.5, sterilized at 121°C for 20 minutes), cultured at 30°C at 160 rpm for 24 hours to activate the strains;

[0036] (2) Expansion of bacterial strain cells

[0037] Add the seed liquid containing 10% (v / v) into the liquid medium (mannitol, glucose, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH5.5, sterilized at 121°C for 20min ) in a shaker culture at 25°C and 150r / min or static culture for 48 hours before use;

[0038] (3) Adding vanillin to prepare vanillyl alcohol

[0039] When a large amount of flocs are formed in the culture medium, add the sterilized and filtered vanilla solution to the culture medium to make the final concentration of vanilla in the culture medium reach 3.0...

technical field

[0001] The invention belongs to the field of preparation of vanillyl alcohol, in particular to a method for preparing vanillyl alcohol by reducing vanillin with bacteria. Background technique

[0002] Vanillyl alcohol, also known as vanillyl alcohol, vanillic alcohol, 4-hydroxy-3-methoxybenzyl alcohol, has a molecular weight of 154.16 and has a creamy sweet and coconut aroma. Compared with vanillin, the aroma is softer, longer-lasting, and more stable in nature. Vanillic alcohol is a recognized fragrance in the world and is suitable for almost all fragrance types. It can be widely used in food, tobacco and wine, beverages and daily chemical products, with unique and outstanding sweetness and aroma.

[0003] It generally exists in the tubers of Gastrodia elata Bl., an orchid plant, and is the active ingredient in Gastrodia elata. In terms of drug efficacy, it has obvious sedative effect, can reduce the spontaneous activity of mice, can prolong the sleep tim...

Examples

Embodiment 1

[0018] (1) Activated bacteria

[0019] Take a ring of Acetobacter xylinum ATCC23770 from the agar slant medium where the strains are preserved, and insert it into the liquid seed medium (mannitol, glucose, sucrose or glycerin 25g, peptone 3g, yeast extract 5g, water 1L, pH3 .5, sterilized at 121°C for 20 minutes), cultured at 160 rpm at 30°C for 12 hours to activate the bacteria;

[0020] (2) Expansion of bacterial strain cells

[0021] Add the seed liquid containing 6% (v / v) into the liquid medium (glucose, mannitol, sucrose or glycerin 25g, peptone 3g, yeast extract 5g, water 1L, pH3.5, sterilized at 121°C for 20min) medium, cultured on a shaking table at 20°C and 100r / min or statically cultured for 12 hours before use;

[0022] (3) Adding vanillin to prepare vanillyl alcohol

[0023] After a large amount of flocs are formed in the culture medium, add the sterile filtered vanillin solution to the culture medium respectively, so that the final concentration of vanillin in ...

Embodiment 2

[0027] (1) Activated bacteria

[0028] Take a ring of Acetobacter xylinum ATCC23770 from the agar slant medium where the strains are preserved, and insert it into the liquid seed medium (mannitol, glucose, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH 5.5, sterilized at 121°C for 20 minutes), cultured at 160 rpm at 30°C for 18 hours to activate the strains;

[0029] (2) Expansion of bacterial strain cells

[0030] Add the seed liquid containing 8% (v / v) into the liquid medium (glucose, mannitol, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH5.5, sterilized at 121°C for 20min ) in a shaker culture at 30°C and 250r / min or static culture for 18 hours before use;

[0031] (3) Adding vanillin to prepare vanillyl alcohol

[0032] After a large amount of flocs are formed in the culture medium, add the sterile filtered vanillin solution to the culture medium respectively, so that the final concentration of vanillin in the culture medium ...

Embodiment 3

[0034] (1) Activated bacteria

[0035] Take a ring of Acetobacter xylinum ATCC23770 from the agar slant medium where the strains are preserved, and insert it into the liquid seed medium (mannitol, glucose, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH 5.5, sterilized at 121°C for 20 minutes), cultured at 30°C at 160 rpm for 24 hours to activate the strains;

[0036] (2) Expansion of bacterial strain cells

[0037] Add the seed liquid containing 10% (v / v) into the liquid medium (mannitol, glucose, sucrose or fructose 25g, tryptone 3g, yeast extract 5g, water 1L, pH5.5, sterilized at 121°C for 20min ) in a shaker culture at 25°C and 150r / min or static culture for 48 hours before use;

[0038] (3) Adding vanillin to prepare vanillyl alcohol

[0039] When a large amount of flocs are formed in the culture medium, add the sterilized and filtered vanilla solution to the culture medium to make the final concentration of vanilla in the culture medium reach 3.0...

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