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Medium for promoting toxin-producing capability of clostridium tetani

A Clostridium and tetanus technology, applied in the direction of bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of low fermentation toxin production capacity, difficult purification, low potency, etc., and achieve the effect of improving toxin production capacity

Inactive Publication Date: 2016-04-13
YUXI WALVAX BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved in the present invention is to overcome the problems that the existing Clostridium tetani culture medium has low fermentation toxin production capacity, difficult purification, low yield and low titer during use.

Method used

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  • Medium for promoting toxin-producing capability of clostridium tetani
  • Medium for promoting toxin-producing capability of clostridium tetani
  • Medium for promoting toxin-producing capability of clostridium tetani

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0047] (3) Preparation of need testing solution: get need testing 15ml, add step (3) (1) described borate buffer 15ml, carry out 2 times dilution to obtain 2 times dilution.

[0048] Assay method: Accurately measure 0.3ml, 0.4mL, 0.5ml, 0.6ml, 0.7ml of the standard solution described in step (3) (2), add flocculent reaction tubes respectively, and each flocculent reaction tube adds step (three) ) (3) 1ml of the test solution, sow evenly, place in a water bath at 45-50°C, observe continuously, and record the order and time of flocculent appearance. Take another 5 flocculent reaction tubes, repeat the above test, put the first flocculent tube in the middle, and add different amounts of the standard solution described in step (3) (2) to the front and rear tubes, with an interval of 0.05ml between each tube , that is, each tube has a difference of 0.05ml, and then 1ml of the test solution described in step (3) (3) is added to each tube to observe the occurrence of floc. According...

Embodiment 2— Embodiment 7

[0053] Embodiment 2—Example 7 Except that the content of the materials listed in Table 4 is different, all the other components and their content and preparation, No. II factor formula and preparation, No. I factor formula and preparation, and all other operations in the medium are all the same as Embodiment 1 is the same and will not be repeated here. The results are shown in Table 5.

[0054] In table 4 embodiment 2-embodiment 7 and embodiment 1

[0055] List of differences in component content

[0056]

[0057]

[0058] Table 5 embodiment 1-embodiment 7 test product floc unit

[0059]

[0060] Above 7 embodiment results show: the medium formula of embodiment 7 is the formula most suitable for Clostridium tetani fermentation, so embodiment 7 is repeated verification, following embodiment 8, embodiment 9, embodiment 10 are Repeat verification.

[0061] The culture medium formula of embodiment 8-embodiment 10 is the same as that of embodiment 7, which is a verific...

Embodiment 1

[0063] Embodiment 11 and embodiment 12 are based on the formula of embodiment 7, and the content of each component has the ± variation described in the technical solution of the present invention, and all the other operations are the same as embodiment 1, and are not repeated. See Table 6, Table 7 and Table 8 for the formulas of No. 1 factor, No. 2 factor and culture medium in Example 11 and Example 12. The experimental results are shown in Table 9.

[0064] No. I factor formula in table 6 embodiment 7, embodiment 11, embodiment 12

[0065]

[0066] No. II factor formula in table 7 embodiment 7, embodiment 11, embodiment 12

[0067]

[0068] Medium formula in table 8 embodiment 7, embodiment 11, embodiment 12

[0069]

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Abstract

The invention discloses a medium for promoting toxin-producing capability of clostridium tetani. One liter of the medium is prepared from 1.00+ / -0.05 g of beef gastric enzyme digestive juice total nitrogen, 1.00+ / -0.05 g of cheese digestive juice total nitrogen, 5.0+ / -0.1 g of sodium acetate, 1.00+ / -0.05 g of disodium hydrogen phosphate, 1.00+ / -0.05 g monopotassium phosphate, 60+ / -2 ml of glycerinum, 2.5+ / -0.1 g / of glucose, 5.0+ / -0.1 ml of a factor I#, 2.00+ / -0.05 ml of a factor II#, 5.0+ / -0.1 g of yeast extract powder, and 0.0025+ / -0.001g of FeCl3.6H2O. The toxin-producing capability is improved by 38-48 Lf / ml, and the protein yield is increased by 36%.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium for improving the toxin-producing ability of Clostridium tetani. Background technique [0002] Clostridium tetani toxin-producing medium is one of the key factors in the preparation process of tetanus toxoid, so the quality of the medium directly affects the quality and yield of tetanus toxoid vaccine. [0003] "Manufacture and Application of Microbial Medium" edited by Chen Tianshou, the first edition in 1995 states that iron (Fe) is a component of catalase, peroxidase, cytochrome, and cytochrome oxidase. The synthesis of these enzymes will be affected, and the growth of vegetative bacteria (Clostridium tetani) also requires the most suitable concentration of iron to be 0.5-0.6mg / L. Iron is also important for bacterial toxins, which can only be produced in a certain concentration range. For example, diphtheria bacteria have the highest amount of toxin ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/145
CPCC12N1/20
Inventor 黄镇李子财施競林建祥
Owner YUXI WALVAX BIOTECH CO LTD
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