A method for preparing monosialotetrahexosyl ganglioside through biotransformation

A technology for ganglioside and monosialic acid, which is applied in the field of preparing monosialotetrahexosylganglioside by biotransformation method, can solve the problems of increased product pollution, low yield, increased workload and the like, and the steps are simple. efficient effect

Active Publication Date: 2017-06-30
NORTHEAST NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content of GM1 is one ten-thousandth of that in brain tissue. The traditional extraction method is to extract and separate from animal brain tissue through the extraction of organic reagents. The method is complicated and the yield is low.
However, the currently published biotransformation method to prepare GM1 (Patent No. 2004100182805) requires the removal of GM1 from the original ganglioside before transformation. Although the total yield has been improved, multiple separations are still required, which increases the workload. , but also increases the risk of product contamination

Method used

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  • A method for preparing monosialotetrahexosyl ganglioside through biotransformation

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Experimental program
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Effect test

Embodiment 1

[0014] The preparation of embodiment 1 fermentation strain

[0015] Take the bacterial strains stored in the cryopreservation tube, inoculate the strains in LB medium, and culture them in shake flasks at 37°C for 1 day. After the strains are cultivated to a certain concentration, they are inoculated in the transformation medium.

Embodiment 2

[0017] Take the ganglioside mixture, add it to 1L transformation medium (inorganic salt, pH 7.0) to make the concentration 10%, add 50ml of the fermentation strain prepared in Example 1, and culture it in a shake flask at 37°C for 3 Take it out two days later, and use the high-performance liquid chromatography test published in the literature Journal ofPharmaceutical and Biomedical Analysis.Vol.8, 1063-1066, 1990. After conversion, GM1 accounts for 70% of the total gangliosides.

Embodiment 3

[0019] Take the ganglioside mixture, add it to 1L transformation medium (inorganic salt, pH 7.0) to make the concentration 20%, add 50ml of the fermentation strain prepared in Example 1, and culture it in a shake flask at 37°C for 3 Take it out two days later, and use the high-performance liquid chromatography test published in the literature Journal of Pharmaceutical and Biomedical Analysis.Vol.8, 1063-1066, 1990. After conversion, GM1 accounts for 80% of the total gangliosides.

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Abstract

The invention discloses a method for biologically transforming polysialic acid four-hexose gangliosides into single sialic acid four-hexose gangliosides (GM1). A polysialic acid four-hexose ganglioside mixture is simply and efficiently hydrolyzed through bacteria for preparing the GM1. The using bacteria belong to the fiber genus, are named as Cellulosimicrobium sp.21, and are preserved in a common microorganism center of the China Committee for Culture Collection of Microorganisms; and the preservation serial number is CGMCC No.7587. The method has the beneficial effects of being good in selectivity, high in activity, simple in process, short in production path, high in product purity and the like; and the possibility is provided for mass production of the GM1.

Description

technical field [0001] The invention relates to a biotransformation method for preparing monosialotetrahexosyl ganglioside, which belongs to the field of biopharmaceuticals. Background technique [0002] Ganglioside is a kind of glycosphingolipid, which is composed of a long-chain sphingosine molecule, a long-chain fatty acid molecule and an oligosaccharide chain containing sialic acid. It mainly exists in the brain and nerve tissues. Gangliosides have a variety of biological activities, play an important role in the process of neurogenesis, growth, and differentiation, and also play an important role in nerve repair after injury, promoting nerve regeneration, promoting nerve axon growth, synapse formation, and recovery Innervation function, improving nerve conduction, promoting the recovery of EEG activity and other neurophysiological indicators; protecting cell membranes, restoring various enzymes of primary cell membranes, etc. In recent years, monosialotetrahexosyl gang...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/44C12R1/01
Inventor 及莉周义发冷佳益孙成新台桂花
Owner NORTHEAST NORMAL UNIVERSITY
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