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Method for preparing gardenia blue pigment by utilizing immobilized yeast cells

A technology for immobilizing yeast and gardenia blue pigment, applied in biochemical equipment and methods, methods based on microorganisms, immobilized on/in organic carriers, etc., can solve the problem of reducing enzyme activity, quality and purity of gardenia blue Unsatisfactory, difficult separation of yeast and products, etc., to achieve the effect of increasing stability, strong anti-pollution ability, and improving quality and purity

Active Publication Date: 2012-07-11
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The method for producing gardenia blue in the prior art mainly adopts geniposide, amino acid and the saccharomycete producing β-glucosidase, and the yeast of the saccharomycete producing β-glucosidase is directly dropped into the geniposide solution and the amino acid solution In the process, the geniposide produced by β-glucosidase hydrolysis reacts with glutamic acid to generate gardenia blue pigment solution. Since there are not only β-glucosidase but also other substances in the yeast, it is directly In the geniposide solution and amino acid solution, impurities will be introduced, and it is difficult to separate the yeast from the product; on the other hand, the enzyme leaves the yeast cell and is directly exposed to the liquid, which reduces the activity of the enzyme. Therefore, the gardenia obtained by this method Blue quality and purity are not ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] a) Inoculate 3mL of β-glucosidase-producing yeast strain into a Erlenmeyer flask containing 100mL of YPDL liquid medium, and ferment and culture it in a shaker at 28°C and 180r / min for 72 hours to obtain a bacterial liquid. After the bacterial liquid is centrifuged, take Add the lower layer of bacterial solution to 2% sodium alginate solution and mix well to form a sodium alginate-yeast suspension, drop the sodium alginate-yeast suspension into a 1mol / L calcium chloride solution to solidify, Form immobilized pellets with a diameter of 1-2mm, the curing temperature is 20°C, and the curing time is 10h; the immobilized pellets are placed in 0.01mol / L calcium chloride solution to harden to form immobilized yeast cells, and the hardening time is 1h; store the prepared immobilized yeast cells at 4°C for later use;

[0033] b) Weigh 100g gardenia powder and dissolve it in 1L distilled water, filter, add the filtrate to a column composed of polystyrene non-polar macroporous ads...

Embodiment 2

[0036] a) Inoculate 5mL of β-glucosidase-producing yeast strains into a Erlenmeyer flask containing 100mL of YPDL liquid medium, ferment and culture them in a shaker at 28°C and 180r / min for 72 hours to obtain bacterial liquid, and after centrifugation, Take the lower layer of bacterial solution and add it to 4% sodium alginate solution and mix well to form a sodium alginate-yeast suspension, and drop the sodium alginate-yeast suspension into a 3mol / L calcium chloride solution to solidify , forming immobilized pellets with a diameter of 1-2mm, the curing temperature is 30°C, and the curing time is 15h; the immobilized pellets are placed in 0.05mol / L calcium chloride solution to harden to form immobilized yeast cells, and the curing time is for 3 hours; store the prepared immobilized yeast cells at 4°C for later use;

[0037] b) Weigh 200g gardenia powder and dissolve it in 5L distilled water, filter, add the filtrate to a column composed of polystyrene type non-polar macroporo...

Embodiment 3

[0040] a) Inoculate 7mL of β-glucosidase-producing yeast strain into a Erlenmeyer flask containing 100mL of YPDL liquid medium, and ferment and culture it in a shaker at 28°C and 180r / min for 72 hours to obtain a bacterial liquid. After the bacterial liquid is centrifuged, take Add the lower layer of bacterial solution to 6% sodium alginate solution and mix well to form a sodium alginate-yeast suspension, drop the sodium alginate-yeast suspension into a 4mol / L calcium chloride solution to solidify, Form immobilized pellets with a diameter of 1-2 mm, the curing temperature is 37°C, and the curing time is 18 hours; the immobilized pellets are placed in 0.1mol / L calcium chloride solution to harden to form immobilized yeast cells, and the hardening time is 5h; store the prepared immobilized yeast cells at 4°C for later use;

[0041] b) Weigh 200g gardenia powder and dissolve it in 10L distilled water, filter, add the filtrate to a column composed of polystyrene type non-polar macr...

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Abstract

The invention relates to a method for preparing gardenia blue pigment by utilizing immobilized yeast cells. The method comprises the following steps of: a) inoculating a yeast strain for producing beta-glucuroide to a culture medium, fermenting to obtain a bacterial liquid, centrifuging the bacterial liquid, embedding a substratum bacterial liquid by using a sodium alginate solution to form a sodium alginate-saccharomycete suspension, and dripping the sodium alginate-saccharomycete suspension into a calcium chloride solution dropwise to form the immobilized yeast cells; b) stirring and mixing a geniposide solution, an amino acid solution and the immobilized yeast cells to obtain a gardenia blue solution; and c) adding the gardenia blue solution into a column which consists of macroporous alkalescent styrene anion exchange resin, eluting by using an ethanol solution, collecting eluent, concentrating the eluent, and spray-drying a concentrated solution to obtain gardenia blue powder. According to the method, a saccharomycete fermentation liquid is immobilized directly without treatment, so that the problems of difficulty in separation of yeast and products, high possibility of loss and the like by the conventional method for performing fermentation by free yeast are solved, and the purity of the gardenia blue is improved.

Description

technical field [0001] The invention relates to a method for preparing natural pigment, in particular to a method for preparing gardenia blue pigment by using immobilized yeast cells. Background technique [0002] At present, there are hundreds of kinds of natural pigments developed in various countries in the world, and more than 40 kinds have been approved for use in my country, but most of them are red and yellow, and the output of natural blue pigments in my country is not high. Gardenia blue pigment is a kind of safe and non-toxic edible natural pigment made from the fruit of the Rubiaceae plant Gardenia jasminoides by using bioengineering technology. It is heat-resistant, light-resistant, acid-resistant and alkali-resistant, has a wide range of pH applications, and is easy to dissolve Suitable for water and low alcohol, it can be used alone instead of chemically synthesized blue pigment, and can also be mixed with red and yellow pigments to form green, purple, brown an...

Claims

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Application Information

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IPC IPC(8): C12P17/16C12N11/10C12N11/04C12R1/865
Inventor 李忠卜素居荣华马飞飞季红峰卢锦峰
Owner NANJING FORESTRY UNIV
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