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Method for producing inositol by fermenting recombinant escherichia coli

A technology for recombining Escherichia coli and Escherichia coli, which is applied in the fields of biological fermentation engineering and enzyme engineering, can solve the problem of low yield and achieve the effect of reducing the production and accumulation of acetic acid

Active Publication Date: 2022-01-21
山东福洋生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The yield of this method is low, and ethanol and boric acid are added during the operation, which has certain risks.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A method for recombinant Escherichia coli fermentation to produce inositol, the specific steps are as follows:

[0039] (1) 150mL shake flask seed preparation

[0040]Prepare primary seed medium (primary seed medium formula: peptone 1.5g, yeast powder 0.75g, sodium chloride 1.5g), 50μg / ml kanamycin (final concentration), pH7.0, 121°C, incinerated Bacteria 30min.

[0041] Inoculate recombinant Escherichia coli with 1% inoculum amount, culture at 35° C., 220 rpm, and shaker for 5 hours to obtain a first-grade seed solution.

[0042] (2) Measure the OD of the primary seed solution with an absorbance photometer 600 When =3.0, transfer to secondary seed medium (glucose 30g / L, potassium dihydrogen phosphate 10g / L, ammonium chloride 3g / L, sodium chloride 5g / L, heptahydrate sulfuric acid Magnesium 0.247g / L, calcium chloride 0.011g / L) in the 5L secondary seed tank; secondary seed tank control reaction temperature 35 ℃, dissolved oxygen (DO) 70-80%, rotating speed is 200rpm / mi...

Embodiment 2

[0059] A method for recombinant Escherichia coli fermentation to produce inositol, the specific steps are as follows:

[0060] (1) 150mL shake flask seed preparation

[0061] Prepare primary seed medium (primary seed medium formula: peptone 1.5g, yeast powder 0.75g, sodium chloride 1.5g), 50μg / ml kanamycin (final concentration), pH 7.0, 121°C, sterilized 30min.

[0062] Inoculate recombinant Escherichia coli with 1% inoculum amount, culture at 35° C., 220 rpm, and shaker for 5 hours to obtain a first-grade seed solution.

[0063] (2) Measure the OD of the primary seed solution with an absorbance photometer 600 When =3.0, transfer to secondary seed medium (glucose 30g / L, potassium dihydrogen phosphate 10g / L, ammonium chloride 3g / L, sodium chloride 5g / L, heptahydrate sulfuric acid Magnesium 0.247g / L, calcium chloride 0.011g / L) in the 5L secondary seed tank; secondary seed tank control reaction temperature 35 ℃, dissolved oxygen (DO) 70-80%, rotating speed is 200rpm / min, pH 7....

Embodiment 3

[0080] A method for recombinant Escherichia coli fermentation to produce inositol, the specific steps are as follows:

[0081] (1) 150mL shake flask seed preparation

[0082] Prepare primary seed medium (primary seed medium formula: peptone 1.5g, yeast powder 0.75g, sodium chloride 1.5g), 50μg / ml kanamycin (final concentration), pH7.0, 121°C, incinerated Bacteria 30min.

[0083] Inoculate recombinant Escherichia coli with 1% inoculum amount, culture at 35° C., 220 rpm, and shaker for 5 hours to obtain a first-grade seed solution.

[0084] (2) Measure the OD of the primary seed solution with an absorbance photometer 600 When =3.0, transfer to secondary seed medium (glucose 30g / L, potassium dihydrogen phosphate 10g / L, ammonium chloride 3g / L, sodium chloride 5g / L, heptahydrate sulfuric acid Magnesium 0.247g / L, calcium chloride 0.011g / L) in the 5L secondary seed tank; secondary seed tank control reaction temperature 35 ℃, dissolved oxygen (DO) 70-80%, rotating speed is 200rpm / m...

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Abstract

The invention discloses a method for producing inositol by fermenting recombinant escherichia coli, and belongs to the technical field of biological fermentation engineering and enzyme engineering. The invention discloses a method for producing inositol by fermenting recombinant escherichia coli, the recombinant escherichia coli is used as a starting strain, an OUR and RQ feedback regulation fermentation mechanism is established, and the method specifically comprises the following steps of: regulating the fed-batch feeding rate according to the OUR change trend of a thallus index growth stage at the early stage of fermentation and a thallus survival stage at the later stage of fermentation; according to the change of the RQ value, the glucose feeding rate can be quickly adjusted, the generation and accumulation of acetic acid in the fermentation liquor are reduced, the influence of acetic acid on the enzyme production of escherichia coli is reduced, and the conversion rate of inositol is improved. According to the invention, the concentration of inositol produced in fermentation liquor is 110-130 g / L, glucose is converted into inositol, the conversion rate reaches 70%, an inositol finished product is crystallized, and the yield is 70%.

Description

technical field [0001] The invention relates to the technical fields of biological fermentation engineering and enzyme engineering, and more specifically relates to a method for producing inositol by recombinant Escherichia coli fermentation. Background technique [0002] Inositol, also known as cyclohexyl hexyl alcohol, hexahydroxycyclohexane, cyclohexitol, myo-inositol, and inositol, is one of the B vitamins. Due to the different orientation of the hydroxyl group relative to the ring plane, there are 9 isomers in total. Body, of which 7 are non-optical and 2 are optically active (levorotatory and dextrorotatory). Inositol is widely distributed in animals and plants, and is a growth factor for animals and microorganisms. It is mainly used to treat liver cirrhosis, hepatitis, fatty liver, and high blood cholesterol. Because inositol is an essential substance for the growth of humans, animals and microorganisms, it has important applications in feed, medicine, food and othe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/02C12Q3/00C12R1/19
CPCC12P7/02C12Q3/00
Inventor 不公告发明人
Owner 山东福洋生物科技股份有限公司
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