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Bacillus, malodor pseudomonads and their use in splitting amygdalinic acid raceme

A technology of Pseudomonas putida and racemate, applied in bacteria, fermentation and other directions, can solve the problems of long reaction time, potential safety hazards, cumbersome splitting steps, etc., and achieve the effect of low production cost

Inactive Publication Date: 2006-01-04
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The technology described by this patented inventor allows for easy access to highly pure mundelsuccinate without expensive methods like chemical synthesis. This results in low costs compared to previous processes while still producing very good quality products at competitive rates.

Problems solved by technology

This patented technology describes methods for making specific types of chemical compounds from certain molecules called mandelsidium acids. These chemistry can include drugs like amoxaparin, which blocks blood clots caused by platelets activated during hemorrhage treatment. However, it may also block other biological processes such as cell growth on nerve tissue. To solve these issues, they have been developed specifically targeting particular metabolites involved in inflammatory reactions.

Method used

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Embodiment 1

[0029] Screening of strains

[0030] Get a small amount of soil dilution supernatant and smear plate (containing enrichment medium, its composition is: benzoylformic acid 2.0g, (NH 4 ) 2 SO 4 2.0g, K 2 HPO 4 1.0g, NaCl 1.0g, MgSO 4 0.5g, 20g of agar, 1000ml of tap water, pH7.0), culture upside down at 25~40℃ for 2~4 days, culture single bacteria on rich medium plate for 1~2 days, then inoculate single colonies into liquid fermentation Medium (glycerol 15.0g, peptone 5.0g, yeast extract 5.0g, K 2 HPO 4 1.0g, KH 2 PO 4 1.0g, NaCl 1.0g, MgSO 4 0.5g, 1000ml of tap water, pH7.0) for 24 hours, and centrifuged to collect bacterial strains Bacillus sp.ECU1008 or Pseudomonas putida ECU1009.

Embodiment 2

[0032] Bacillus sp.ECU1008 (CGMCC No.1387) strain was inoculated into 40ml growth medium (glucose 1.5wt%, peptone 0.5wt%, yeast extract 0.5wt%, K 2 HPO 4 0.1wt%, KH 2 PO 4 0.1wt%, NaCl 0.1wt%, MgSO 4 0.05wt%, the rest is water, pH 7.0), after pre-cultivation on a shaker at 30°C and 160rpm for 12 hours, add 0.5% racemic mandelic acid as an inducer, and continue to cultivate for 12 hours; use this culture solution as a seed, press 10v / v% of the inoculum size was inoculated into 40ml transformation medium (racemic mandelic acid 20.0g, corn steep liquor 1.0g, NH 4 NO 3 , K 2 HPO 4 1.0g, KH 2 PO 4 1.0g, NaCl1g, MgSO 4 0.5g, water 1000ml, pH7.0), 30 ℃, 160rpm shaking table fermentation transformation for 2 days. After the reaction, centrifuge at 10,000×g for 10 min, collect the supernatant, remove most of the water by rotary evaporation, acidify the concentrate with 50% sulfuric acid to pH 1-2, then saturate with salt, extract with ethyl acetate, and dry over anhydrou...

Embodiment 3

[0034] Pseudomonas putida ECU1009 (CGMCC No.1388) bacterial strain was inoculated into 40ml nutrient medium (glucose 1.5wt%, peptone 0.5wt%, yeast extract 0.5wt%, K 2 HPO 4 0.1wt%, KH 2 PO 4 0.1wt%, NaCl0.1wt%, MgSO 4 0.05wt%, the rest is water, pH7.0), pre-cultivated on a shaker at 30°C and 160rpm for 12 hours, added 0.5% mandelic acid as an inducer, and continued to cultivate for 12 hours. Inoculate to 40ml transformation medium (racemic mandelic acid 10.0g, corn steep liquor 1.0g, NH 4 NO 3 , K 2 HPO 41.0g, KH 2 PO 4 1.0g, NaCl1g, MgSO 4 0.5 g, 1000 ml of tap water, pH 7.0), cultured on a shaker at 160 rpm at 28° C. for 2 days. After the reaction, centrifuge at 10,000×g for 10 minutes to remove the bacteria, keep the supernatant, add 1 g of activated carbon to decolorize by heating, remove the activated carbon by filtration, remove most of the water by rotary evaporation under reduced pressure, and acidify the concentrated solution to pH1 with concentrated su...

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Abstract

The present invention discloses one strain of bacillus, ECU1008 (CGMCC No. 1387), specially for degrading (S)-amygdalinic acid, and one strain of Pseudomonas putida, ECU1009 (CGMCC No. 1388), specially for degrading (R)-amygdalinic acid and their use. The strains of the present invention is used in resolving amygdalinic acid raceme, and has the advantages of simple and safe operation, low cost, easy purification of target product, environment friendship, etc.

Description

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Claims

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Application Information

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Owner EAST CHINA UNIV OF SCI & TECH
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