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Phenol-degrading bacteria and method for preparing indigo by conversing indole

A phenol-degrading bacteria and phenol-degrading technology, applied in the field of bioengineering, can solve the problems of ignoring advantages, limited research, and single enzyme resources, and achieve the effects of simple extraction steps, high product concentration, and short production cycle

Active Publication Date: 2014-04-23
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Existing studies on the preparation of indigo involve relatively single enzyme resources, mainly focusing on naphthalene dioxygenase, toluene dioxygenase, styrene monooxygenase and P450 enzymes, etc., and the practical application research is limited; most Most of the research is to use genetic engineering technology to transfer related synthetic genes for recombination to construct engineering bacteria. There are few studies on the use of wild-type strains to synthesize indigo, ignoring the advantages of natural native strains to synthesize indigo

Method used

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  • Phenol-degrading bacteria and method for preparing indigo by conversing indole
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  • Phenol-degrading bacteria and method for preparing indigo by conversing indole

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1, using the phenol-degrading bacteria Pseudomonas monteilii QM to biotransform indole to produce indigo, and investigate the influence of some parameters on the concentration of indigo.

[0025] The steps and parameters for the expanded cultivation and biotransformation of indole to produce indigo by the phenol-degrading bacteria Pseudomonas monteilii QM are as follows:

[0026] 1. Inoculate the phenol-degrading bacteria Pseudomonas monteilii QM strain on a solid plate LB medium (yeast powder 5g / L, peptone 10g / L, NaCl 5g / L, agar 15-20g / L, adjust the pH to 7.0-7.2), Cultivate statically at 37°C for 2 days, store in refrigerator at 4°C, and serve as activated seeds on the plate;

[0027] 2, use the plate activation seed of step 1 gained, be inoculated in the liquid M9 substratum (Na 2 HPO 4 12H 2 O 17.1g / L, KH 2 PO 4 3.0g / L, NH 4 Cl 1.0g / L, NaCl 0.5g / L, FeCl 3 0.00075g / L, 1g / L glucose, 2mmol / L MgSO4), shaking culture at 20-40°C, 50-300r / min for 12-16 hou...

Embodiment 2

[0035] Example 2, phenol-degrading bacteria Pseudomonas monteilii QM biotransforms indole to produce indigo.

[0036] Steps 1-4 are the same as in Example 1, and the dry weight concentration of the expanded cultured Pseudomonas monteilii QM bacteria obtained in step 3 is controlled to be 4.0 g / L.

[0037] In a 50mL Erlenmeyer flask, add 20mL of the expanded cultured Pseudomonas monteilii QM bacteria prepared in step 3, and 15 μL of the indole / acetone solution prepared in step 4, so that the concentration of indole in the reaction solution is 75 mg / L. , 300r / min shaking for 6 hours.

[0038] According to the measurement method described in Example 1, the indigo concentration recorded is 35.24 mg / L.

Embodiment 3

[0039] Example 3, phenol-degrading bacteria Pseudomonas monteilii QM biotransforms indole to produce indigo.

[0040] Steps 1-4 are the same as in Example 1, and the dry weight concentration of the expanded cultured Pseudomonas monteilii QM bacteria obtained in step 3 is controlled to be 8.0 g / L.

[0041] In a 50mL Erlenmeyer flask, add 20mL of the enlarged cultured Pseudomonas monteilii QM bacteria prepared in step 3, 20 μL of the indole / acetone solution prepared in step 4, so that the concentration of indole in the reaction solution is 100mg / L, at 40°C , 250r / min shaking reaction for 8 hours.

[0042] According to the measurement method described in Example 1, the indigo concentration recorded is 31.20 mg / L.

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Abstract

The invention belongs to the biology engineering technical field, and relates to a method for preparing indigo by conversing indole through phenol-degrading bacteria. The invention is characterized in that the phenol-degrading microbial strain capable of effectively catalyzing indole to synthesize indigo is employed, and is Pseudomonas monteilii QM, and the preservation number is CGMCC No.5054; when the biotransformation reaction is carried out, the cell dry weight concentration of the Pseudomonas monteilii QM bacterium in a reaction solution is 0.1-10g / L, the indole concentration is 25-200mg / L, under temperature of 20-40 DEG C, the shaking table rotating speed is controlled with 50-300r / min, and the oscillation reaction lasts for 1-8 hours. The condition of the biotransformation reaction is mild, the product concentration reaches as high as 42.5mg / L, the production period is short, the cost is low, the extraction step is simple, the production step is clean, and the method is suitable for industrial production and application.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a method for preparing indigo by transforming indole with a wild-type phenol-degrading bacterium. Background technique [0002] Indigo is an ancient pigment widely used in industries such as food, medicine, printing and dyeing, and cosmetics. The raw materials, catalysts, and intermediate products of its chemical synthesis are all toxic to a certain extent, causing harm to humans and the environment. [0003] In 1983, Ensley et al. began to use biological methods to synthesize indigo. In 1986, Mermod N et al. cloned the TOL plasmid pWW0 of Pseudomonas putida mt-2, which degrades toluene, xylene or other derivatives of toluene, into Escherichia coli K-12, and obtained the genetic engineering Bacteria can catalyze indole to directly produce 3-oxindole to produce indigo. [0004] There are few reports on the synthesis of indigo by wild bacteria. In 1997, Doukyu et al. used the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P17/16C12R1/38
Inventor 曲媛媛马桥许炳雯张旭旺李新亮周豪孔春雷曹湘禹周集体沈娥
Owner DALIAN UNIV OF TECH
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