Method of producing vanillin by transforming isoeugenol via Saccharomyces cerevisiae
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A technology for isoeugenol and yeast, which is applied in the field of yeast transforming isoeugenol to produce vanillin, can solve the problems such as no yeast disclosure and report, and achieve the effect of satisfying the demand
Active Publication Date: 2016-01-06
YOUGE TIANCHENG BIOTECH YIWU CO LTD
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At present, there are no publications or reports on the conversion of isoeugenol to vanillin by yeast
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preparation example Construction
[0059] 2. Preparation of culture medium
[0060] Raw material preparation:
[0061] 10*YNB (13.4% yeast nitrogen source base medium, containing ammonium sulfate without amino acid): Dissolve 134g of YNB containing ammonium sulfate in 1000ml of water, heat the solution to completely dissolve YNB in water, filter and sterilize at 4°C Store in the refrigerator.
[0062] 500*B (0.02% biotin): Dissolve 20 mg of biotin in 100 ml of water, sterilize by filtration, and store in a refrigerator at 4°C.
[0063] 10*D (20% glucose): Dissolve 200g of D-glucose in 1000ml of water, and sterilize in an autoclave for 15 minutes or by filtration.
[0064] 10*M (5% methanol): Mix 5ml methanol with 95ml water, filter sterilize, and store in a refrigerator at 4°C.
[0065] 1. Preparation of YPD liquid medium (1 liter) (including: 1% yeast extract, 2% peptone, 2% glucose):
[0066] Dissolve 10 g of yeast extract and 20 g of peptone in 900 ml of water. (If it needs to be made into a slant or ...
Embodiment 1
[0078] In a 50mL Erlenmeyer flask, add 0.6g of isoeugenol, 10mL of fermentation broth, add 10mL of glycine / NaOH buffer solution with a pH of 10.4 and a glycine concentration of 200mM, cover the mouth with two layers of gauze, and incubate at 30°C and 200rpm The transformation was carried out by shaking on a shaking table for 48 hours, and the concentration of vanillin in the final reaction solution was determined to be 3.5 g / L.
Embodiment 2
[0080] In a 50mL Erlenmeyer flask, add 0.5g of isoeugenol, 10mL of fermentation broth, add 10mL of glycine / NaOH buffer solution with a pH of 10.4 and a glycine concentration of 200mM, cover the mouth with two layers of gauze, and incubate at 30°C and 200rpm The transformation was carried out by shaking on a shaking table for 48 hours, and the concentration of vanillin in the final reaction solution was determined to be 3.25 g / L.
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Abstract
The invention discloses a method of producing vanillin by transforming isoeugenol via Saccharomyces cerevisiae. The method includes: S1, constructing Saccharomyces cerevisiae for producing isoeugenol monooxygenase; S2, inoculating the constructed Saccharomyces cerevisiae to an YPD (yeast extract peptone dextrose) liquid medium for shake culture; S3, centrifugally collecting bacteria, washing the bacteria with normal saline, and subjecting the bacteria to shake culture in an MGY (minimal glycerol) liquid medium; S4, centrifugally collecting bacteria, washing the bacteria with normal saline, subjecting the bacteria to shake culture in a liquid MM (minimal medium) for 3-5 days with OD600>6; replenishing methanol each day, and adjusting pH with ammonia to obtain fermentation liquor; centrifuging the fermentation liquor selectively to collect supernate; S5, adding isoeugenol, the supernate or the fermentation liquor and buffer solution according to a ratio of (0.1-0.6) g:(9-18) mL:(9-18) mL into a reaction container, and allowing shake transformation at 20-30 DEG C for 12-48 hours to obtain the vanillin. The vanillin is industrially produced using Saccharomyces cerevisiae, and the method is applicable to the food field and the like and is satisfactory and safe.
Description
technical field [0001] The invention relates to the technical field of biotransformation, in particular to a method for yeast to transform isoeugenol to produce vanillin. Background technique [0002] Vanillin is one of the most widely used spices in the industry. It is widely used in the food industry. It can be used as the main raw material for aroma modification and fragrance fixation in food, toothpaste, soap, and tobacco; it can be used as an important raw material or Intermediates, which can be used in the manufacture of commonly used drugs for the treatment of hypertension, heart disease, skin diseases, and elimination of bad breath and diuresis; in the chemical industry, they can be used as chemical additives, anti-hardening agents for plastic products, and metals such as Ni, Cr, and Cd Plating brightener; in agricultural production, vanillin can be used as a crop yield increasing agent and ripening agent, and used to prepare herbicides and insect attractants, etc., ...
Claims
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Application Information
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