Method for increasing yield of guanosine produced by fermenting bacillus subtilis

Abstract

The invention discloses a method for increasing yield of guanosine produced by fermenting bacillus subtilis. The method comprises two major technological processes, namely shaking culture of liquid strain and preparation of guanosine fermentation liquor. In the method, preserved strain GDGR-1006 with stable heredity is selected and used; a novel process of adding malic acid and adding glucose liquid and the malic acid in a flowing mode is adopted during fermentation; and thus, the defects of backwardness of the conventional process in the breeding technology, disproportionality of C (Carbon) to N (Nitrogen) of a fermentation medium, low yield of the guanosine from the strain, long fermentation period, low conversion rate and the like are overcome, the strain has high synchronous productivity, high guanosine production level and high guanosine production level by fermentation, the yield of the guanosine can reach over 45g / L, and the conversion rate is over 28 percent.

Description

(1) Technical field

[0001] The invention belongs to a production method of guanosine, an important intermediate of food and medical products. (2) Background technology

[0002] Guanosine (trade name: GR) is also known as 9-β-D-ribofuranosylguanine, referred to as Guanosine, and its English name is Guanosine. Guanosine is white crystalline powder or off-white crystalline powder, soluble in water and pyrimidine, insoluble in alcohol. Molecular formula is C 10 h 13 h 5 o 5 , the relative molecular weight is 283.24. The chemical structural formula is as follows:

[0003]

[0004] Guanosine has a wide range of uses and is an important intermediate in food and pharmaceutical products. In the field of pharmaceutical industry, guanosine is mainly used as an important intermediate for the synthesis of ribavirin, acyclovir and other nucleoside antiviral drugs, and is also used to manufacture acyclic guanosine (Acyclovir), triazine Ribavirin (ATC), sodium guanosine triphosph...

Examples

example 1

[0032] 1. Take 1g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium sulfate, 0.1g of malic acid, 2g of yeast powder, 2g of agar, and 94.6g of water to prepare the primary culture medium, put it into an eggplant bottle and wait for 22min at 121°C After steam sterilization, it was left to cool down to 37°C, and the preserved strain Bacillus subtilis GDGR-1006 was inserted into it, and cultured statically at 37°C for 22 hours in a sterile state to obtain primary strains.

[0033] 2. Take 36g of glucose, 2.4g of dipotassium hydrogen phosphate, 1.2g of magnesium sulfate, 1.2g of malic acid, 18g of yeast powder, and 1141.2g of water to make a culture medium for shake flasks, put them into shake flasks and steam at 121°C for 22min. After sterilizing, cool to 37°C, add the culture medium for primary strains obtained in step 1, and cultivate on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 160 rpm, a temperature of 37°C, and a time period ...

example 2

[0037] 1. Take 1g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium sulfate, 0.0g of malic acid, 2g of yeast powder, 2g of agar, and 94.7g of water, and put the prepared primary culture medium into an eggplant bottle for 22min at 121°C After steam sterilization, let it cool down to 37°C, insert the preserved strains, and culture them statically for 22 hours in a sterile state at 37°C to obtain primary strains.

[0038] 2. Take 36g of glucose, 2.4g of dipotassium hydrogen phosphate, 1.2g of magnesium sulfate, 0.0g of malic acid, 18g of yeast powder, and 1142.4g of water. Put the prepared culture medium into the shaker flask and steam it at 121°C for 22min. After sterilizing, cool to 37°C, add the primary strain culture medium obtained in the previous step, and cultivate on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 160 rpm, a temperature of 37°C, and a time period of 14 hours. A bottle of liquid bacteria, the resulting bacteria...

example 3

[0042] 1. Take 1g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium sulfate, 0.2g of malic acid, 2g of yeast powder, 2g of agar, and 94.5g of water, and put the prepared primary culture medium into an eggplant bottle for 22min at 121°C After steam sterilization, let it cool down to 37°C, insert the preserved strains, and culture them statically for 22 hours in a sterile state at 37°C to obtain primary strains.

[0043] 2. Take 36g of glucose, 2.4g of dipotassium hydrogen phosphate, 1.2g of magnesium sulfate, 2.4g of malic acid, 18g of yeast powder, and 1140.0g of water. Put the prepared culture medium into the shaker flask and steam it at 121°C for 22min. After sterilizing, cool to 37°C, add the primary strain culture medium obtained in the previous step, and cultivate on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 160 rpm, a temperature of 37°C, and a time period of 14 hours. Bottle of liquid bacteria, the resulting bacteria m...

(1) Technical field

[0001] The invention belongs to a production method of guanosine, an important intermediate of food and medical products. (2) Background technology

[0002] Guanosine (trade name: GR) is also known as 9-β-D-ribofuranosylguanine, referred to as Guanosine, and its English name is Guanosine. Guanosine is white crystalline powder or off-white crystalline powder, soluble in water and pyrimidine, insoluble in alcohol. Molecular formula is C 10 h 13 h 5 o 5 , the relative molecular weight is 283.24. The chemical structural formula is as follows:

[0003]

[0004] Guanosine has a wide range of uses and is an important intermediate in food and pharmaceutical products. In the field of pharmaceutical industry, guanosine is mainly used as an important intermediate for the synthesis of ribavirin, acyclovir and other nucleoside antiviral drugs, and is also used to manufacture acyclic guanosine (Acyclovir), triazine Ribavirin (ATC), sodium guanosine triphosph...

Examples

example 1

[0032] 1. Take 1g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium sulfate, 0.1g of malic acid, 2g of yeast powder, 2g of agar, and 94.6g of water to prepare the primary culture medium, put it into an eggplant bottle and wait for 22min at 121°C After steam sterilization, it was left to cool down to 37°C, and the preserved strain Bacillus subtilis GDGR-1006 was inserted into it, and cultured statically at 37°C for 22 hours in a sterile state to obtain primary strains.

[0033] 2. Take 36g of glucose, 2.4g of dipotassium hydrogen phosphate, 1.2g of magnesium sulfate, 1.2g of malic acid, 18g of yeast powder, and 1141.2g of water to make a culture medium for shake flasks, put them into shake flasks and steam at 121°C for 22min. After sterilizing, cool to 37°C, add the culture medium for primary strains obtained in step 1, and cultivate on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 160 rpm, a temperature of 37°C, and a time period ...

example 2

[0037] 1. Take 1g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium sulfate, 0.0g of malic acid, 2g of yeast powder, 2g of agar, and 94.7g of water, and put the prepared primary culture medium into an eggplant bottle for 22min at 121°C After steam sterilization, let it cool down to 37°C, insert the preserved strains, and culture them statically for 22 hours in a sterile state at 37°C to obtain primary strains.

[0038] 2. Take 36g of glucose, 2.4g of dipotassium hydrogen phosphate, 1.2g of magnesium sulfate, 0.0g of malic acid, 18g of yeast powder, and 1142.4g of water. Put the prepared culture medium into the shaker flask and steam it at 121°C for 22min. After sterilizing, cool to 37°C, add the primary strain culture medium obtained in the previous step, and cultivate on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 160 rpm, a temperature of 37°C, and a time period of 14 hours. A bottle of liquid bacteria, the resulting bacteria...

example 3

[0042] 1. Take 1g of glucose, 0.2g of dipotassium hydrogen phosphate, 0.1g of magnesium sulfate, 0.2g of malic acid, 2g of yeast powder, 2g of agar, and 94.5g of water, and put the prepared primary culture medium into an eggplant bottle for 22min at 121°C After steam sterilization, let it cool down to 37°C, insert the preserved strains, and culture them statically for 22 hours in a sterile state at 37°C to obtain primary strains.

[0043] 2. Take 36g of glucose, 2.4g of dipotassium hydrogen phosphate, 1.2g of magnesium sulfate, 2.4g of malic acid, 18g of yeast powder, and 1140.0g of water. Put the prepared culture medium into the shaker flask and steam it at 121°C for 22min. After sterilizing, cool to 37°C, add the primary strain culture medium obtained in the previous step, and cultivate on a reciprocating shaker with an inoculum size of 0.1%, a rotation speed of 160 rpm, a temperature of 37°C, and a time period of 14 hours. Bottle of liquid bacteria, the resulting bacteria m...

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