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Process for multistep directional o-cresol conversion with biological method

A biotransformation and directional transformation technology, applied in the field of bioengineering, can solve problems such as difficult to control the transformation process at all levels, difficult to achieve controllable and directional transformation of toluene and methylbenzene, and achieve directional controllable and directional transformation of the production process. The effect of low cost and short production cycle

Inactive Publication Date: 2011-10-26
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This multi-stage biotransformation method is difficult to control the conversion process at each level. During the conversion process, unconverted substrates such as toluene and methylene methylbenzene inhibit the second and third step conversion reactions, and the second step conversion The resulting corresponding alcohols inhibit the third step reaction, and it is difficult to realize the controllable and directional conversion of toluene and partial methylbenzene

Method used

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  • Process for multistep directional o-cresol conversion with biological method
  • Process for multistep directional o-cresol conversion with biological method
  • Process for multistep directional o-cresol conversion with biological method

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Experimental program
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Effect test

Embodiment 1

[0028] Embodiment 1, phenol hydroxylase genetically engineered bacteria PH_ IND and biphenyl exodiol dioxygenase genetically engineered bacteria BphC_LA-4 for multi-stage directional conversion of o-cresol, and the effects of some parameters on the production and conversion rate of 3-methylcatechol were investigated.

[0029] Engineering bacteria PH_ IND The medium used for expanding the culture of engineering bacteria BphC_LA-4 is LB medium, the formula is: 10g / L NaCl, 10g / L peptone, 5g / L yeast extract, adjust the pH to 7.0-7.2, and sterilize at 121°C After 20 minutes, add kanamycin at a final concentration of 10-20 mmol / L as an antibiotic after the culture medium is cooled. The formula of solid plate medium is 15-20g / L agar added on the basis of LB medium. The expanded culture of genetically engineered bacteria and the biotransformation o-cresol reaction steps and parameters are as follows:

[0030] 1. The engineering bacteria PH_ IND Inoculate the solid plate LB medium ...

Embodiment 2

[0045] Embodiment 2, phenol hydroxylase genetically engineered bacteria PH_ IND Multi-stage directed transformation of o-cresol with BphC_LA-4 genetically engineered bacterium BphC_LA-4.

[0046] LB substratum formula and the expanded culture of genetically engineered bacterium and biotransformation o-cresol reaction step 1~5 are with embodiment 1, make the engineering bacterium PH that step 3 makes by controlling the collected thalline quality. IND The dry weight of whole cells is 20g / L, and the dry weight of whole cells of engineering bacteria BphC_LA-4 is 25g / L.

[0047] The first level biotransformation reaction: in the Erlenmeyer flask of 50mL, add the engineering bacteria PH_ that 20mL step 3 makes IND Whole cells, 15 μL of o-cresol solution prepared in step 4, 600 μL of glucose stock solution prepared in step 5, so that the concentration of o-cresol in the reaction solution is 50 mg / L, and the concentration of glucose is 30 mmol / L. At 40 ° C, shake The bed speed is 18...

Embodiment 3

[0050] Embodiment 3, phenol hydroxylase genetically engineered bacteria PH_ IND Multi-stage directed transformation of o-cresol with BphC_LA-4 genetically engineered bacterium BphC_LA-4.

[0051] LB substratum formula and the expanded culture of genetically engineered bacteria and biotransformation o-cresol reaction steps 1~5 are with embodiment 1, wherein the engineering bacteria PH that step 3 makes IND The dry weight of whole cells is 25g / L, and the dry weight of whole cells of engineering bacteria BphC_LA-4 is 20g / L.

[0052] The first level biotransformation reaction: in the Erlenmeyer flask of 50mL, add the engineering bacteria PH_ that 20mL step 3 makes IND Whole cells, 40 μL of o-cresol solution prepared in step 4, 100 μL of glucose stock solution prepared in step 5, so that the concentration of o-cresol in the reaction solution is 200 mg / L, and the concentration of glucose is 5 mmol / L. At 35 ° C, shake Bed speed 200r / min, shaking reaction 240min;

[0053]The second...

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Abstract

The invention which belongs to the technical field of biological engineering relates to a process for o-cresol compound conversion with a biological method. The process is characterized in that: a first step biological conversion reaction is carried out to obtain a product 3-methycatechol with conditions that a dry weight of whole cells of phenol hydroxylase gene engineering bacteria PH-IND in a reaction solution is from 20 to 40 g / L, a concentration of o-cresol is from 50 to 700 mg / L, the concentration of glucose is from 5 to 40 mmol / L, a temperature is from 20 to 40 DEG C, a shaker speed iscontrolled from 50 to 200 r / min, and an oscillation reaction period is from 10 to 240 min; and a second step biological conversion reaction is carried out with the conditions that the dry weight of the whole cells of biphenyl dioxygenase gene engineering bacteria BphC-LA-4 which are added in the reaction solution is from 20 to 40 g / L, the shaker speed is controlled from 50 to 200 r / min, and the oscillation reaction period is from 5 to 50 min. The method has the advantages of mild reaction condition, single product and simple extraction step, allows the product to be recycled, and 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 converting o-cresol compounds by a biological method. Background technique [0002] Cresol compounds are a representative substance among phenolic compounds. They are used in the production of insecticides such as fenitrothion, fenthion, medicarb, and permethrin, as well as color films, resins, and plasticizers. The intermediate synthesized with spices has strong corrosiveness and toxicity, and has been included in the list of environmental priority pollutants by the US EPA. Regarding the treatment method of cresol-containing wastewater, compared with the physical and chemical method, the biological method has the advantages of economy, safety, low treatment threshold, less residue, and no secondary pollution. [0003] At present, in the research of harmless biodegradation of cresol compounds, it is found that many bacteria, fungi, algae and aquatic animals and...

Claims

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Application Information

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IPC IPC(8): C12P7/44C12R1/19
Inventor 曲媛媛时胜男张瑞杰马桥张旭旺周集体周豪许炳雯李新亮孔春雷
Owner DALIAN UNIV OF TECH
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