Method for extracting bilirubin from ox bile by using fermentation process

Abstract

The invention provides a method for extracting bilirubin from ox bile by using a fermentation process. The method mainly comprises the following four steps: fermenting ox bile, sterilizing an ox bile fermentation solution, forming bilirubin, extracting and refining bilirubin. By virtue of the method, high-yield beta-glucuronidase and ox-bile-resistant clostridium perfringens are added into the animal ox bile and fermented; a mercaptoalcohol reducing agent is added, so that the effects of improving the activity of beta-glucuronidase and ox-bile-resistant clostridium perfringens and resisting oxidants are implemented; the overall process is greatly optimized under the control of the special condition; the production efficiency is improved; the cost is reduced; the yield of the finally prepared bilirubin is 0.010-0.014%; and the purify of the bilirubin reaches over 92%.

Description

technical field

[0001] The invention relates to a method for extracting bilirubin, in particular to a method for extracting bilirubin from bovine bile by using a fermentation method. Background technique

[0002] Bilirubin has a variety of pharmacological effects and is the main raw material for the manufacture of artificial bezoar. It is widely used in medicine and is in great demand in the market. However, the current extraction methods of bilirubin are mainly extracted from various animal bile (pig, cattle, sheep), and the content of bilirubin in animal bile is extremely low, the extraction process is cumbersome, and the extraction rate is low (such as Wu The yield of the product obtained by Weiping and others by the direct and rapid extraction method is 0.008%-0.013%, the yield obtained by Shang Wenxue and others by the resin method is 0.006174%, and the yield of the cholecalcium salt method is lower and the production cost is high, resulting in Bilirubin is extremely e...

Examples

Embodiment 1

[0030] (1) The strain of Clostridium perfringens was expanded and cultured in liquid medium for 22 hours to obtain an expanded bacterial solution; the expanded bacterial solution was added to the filtered fresh bovine bile at a temperature of 28 °C Fermentation was carried out for 45h to obtain a fermented liquid, and the pH of the fermented liquid was maintained at 5 during the fermentation process; the volume of the amplified bacterial liquid was 9% of the volume of fresh ox bile;

[0031] (2) adding lysozyme to the fermentation broth obtained in the upper step, carrying out enzymatic hydrolysis for 30 minutes at a temperature of 37-38 degrees, and adjusting its pH to 6.8 to obtain a lysate;

[0032] The mass volume concentration of the lysozyme in the fermentation broth is 0.2g / 1L;

[0033] (3) Add 10% NaOH solution to the lysate obtained in the upper step, and adjust its pH to 11 to obtain a saponification solution;

[0034] (4) Heat and boil the saponified solution obtai...

Embodiment 2

[0046] (1) The strain of Clostridium perfringens was expanded and cultured in liquid medium for 24 hours to obtain an expanded bacterial solution; the expanded bacterial solution was added to the filtered fresh bovine bile at a temperature of 32 °C Fermentation was carried out for 48h to obtain a fermented liquid, and the pH of the fermented liquid was maintained at 6 during the fermentation process; the volume of the amplified bacterial liquid was 11% of the volume of fresh ox bile;

[0047](2) adding lysozyme to the fermentation broth obtained in the upper step, carrying out enzymatic hydrolysis for 50 minutes at a temperature of 37-38 degrees, and adjusting its pH to 7.5 to obtain a lysate;

[0048] The mass volume concentration of the lysozyme in the fermentation broth is 0.25g / 1L;

[0049] (3) Add 10% NaOH solution to the lysate obtained in the upper step, and adjust its pH to 12 to obtain a saponification solution;

[0050] (4) Heat and boil the saponified solution obta...

technical field

[0001] The invention relates to a method for extracting bilirubin, in particular to a method for extracting bilirubin from bovine bile by using a fermentation method. Background technique

[0002] Bilirubin has a variety of pharmacological effects and is the main raw material for the manufacture of artificial bezoar. It is widely used in medicine and is in great demand in the market. However, the current extraction methods of bilirubin are mainly extracted from various animal bile (pig, cattle, sheep), and the content of bilirubin in animal bile is extremely low, the extraction process is cumbersome, and the extraction rate is low (such as Wu The yield of the product obtained by Weiping and others by the direct and rapid extraction method is 0.008%-0.013%, the yield obtained by Shang Wenxue and others by the resin method is 0.006174%, and the yield of the cholecalcium salt method is lower and the production cost is high, resulting in Bilirubin is extremely e...

Examples

Embodiment 1

[0030] (1) The strain of Clostridium perfringens was expanded and cultured in liquid medium for 22 hours to obtain an expanded bacterial solution; the expanded bacterial solution was added to the filtered fresh bovine bile at a temperature of 28 °C Fermentation was carried out for 45h to obtain a fermented liquid, and the pH of the fermented liquid was maintained at 5 during the fermentation process; the volume of the amplified bacterial liquid was 9% of the volume of fresh ox bile;

[0031] (2) adding lysozyme to the fermentation broth obtained in the upper step, carrying out enzymatic hydrolysis for 30 minutes at a temperature of 37-38 degrees, and adjusting its pH to 6.8 to obtain a lysate;

[0032] The mass volume concentration of the lysozyme in the fermentation broth is 0.2g / 1L;

[0033] (3) Add 10% NaOH solution to the lysate obtained in the upper step, and adjust its pH to 11 to obtain a saponification solution;

[0034] (4) Heat and boil the saponified solution obtai...

Embodiment 2

[0046] (1) The strain of Clostridium perfringens was expanded and cultured in liquid medium for 24 hours to obtain an expanded bacterial solution; the expanded bacterial solution was added to the filtered fresh bovine bile at a temperature of 32 °C Fermentation was carried out for 48h to obtain a fermented liquid, and the pH of the fermented liquid was maintained at 6 during the fermentation process; the volume of the amplified bacterial liquid was 11% of the volume of fresh ox bile;

[0047](2) adding lysozyme to the fermentation broth obtained in the upper step, carrying out enzymatic hydrolysis for 50 minutes at a temperature of 37-38 degrees, and adjusting its pH to 7.5 to obtain a lysate;

[0048] The mass volume concentration of the lysozyme in the fermentation broth is 0.25g / 1L;

[0049] (3) Add 10% NaOH solution to the lysate obtained in the upper step, and adjust its pH to 12 to obtain a saponification solution;

[0050] (4) Heat and boil the saponified solution obta...