Preparation method of bacillus spores

A technology of bacillus and spores, which is applied in the field of microbial spore preparation, can solve the problems of time-consuming, cumbersome process, and high production cost, and achieve the effects of shortened culture time, simple operation steps, and good repeatability

Active Publication Date: 2017-02-22
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, in the existing technology, it has been possible to prepare small doses of spores of Bacillus stearothermophilus, but there are many technical obstacles in large-scale mass production, mainly due to the low rate of spore formation and high production costs. Problems, at the same time, it takes a long time and the process is cumbersome

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  • Preparation method of bacillus spores

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Effect test

Embodiment 1

[0021] A preparation method for bacillus spores, comprising the following steps:

[0022] Step 1) Vegetative cell proliferation:

[0023] Inoculate 1g of Bacillus stearothermophilus into 10g of nutrient-enriched medium and culture it for 12 hours at 50°C with a rotational speed of 150rpm / min; then add 90g of nutrient-enriched medium and continue to cultivate for 12 hours to obtain vegetative cells culture medium;

[0024] Step 2) spore transformation:

[0025] Add 1 g of inorganic ionic medium to 1 g of vegetative cell culture medium, and culture at 55°C for 12 hours at a rotational speed of 100 rpm / min; then add 1 g of inorganic ionic medium every 12 hours until the added inorganic ions are cultured The ratio of the total mass parts of the base to the mass parts of the nutrient-rich medium reaches 7:1; a culture solution containing bacillus spores is obtained.

[0026] Among them, the rich nutrient medium preferably includes: 5-20 parts by weight of soybean peptone, 5 part...

Embodiment 2

[0029] A preparation method for bacillus spores, comprising the following steps:

[0030] Step 1) Vegetative cell proliferation:

[0031] Inoculate 1g of Bacillus stearothermophilus into 10g of nutrient-enriched medium and culture it for 12 hours at 55°C with a rotation speed of 250rpm / min; then add 90g of nutrient-enriched medium and continue to cultivate for 12 hours to obtain vegetative cells culture medium;

[0032] Step 2) spore transformation:

[0033] Add 1g of inorganic ionic medium to 1g of vegetative cell culture medium, and culture at 60°C with a rotational speed of 150rpm / min for 12h; then add 1g of inorganic ionic medium every 12h until the added inorganic ionic culture The ratio of the total mass parts of the base to the mass parts of the nutrient-rich medium reaches 10:1; a culture solution containing bacillus spores is obtained.

[0034] Among them, the rich nutrient medium preferably includes: 5-20 parts by weight of soybean peptone, 5 parts by weight of be...

Embodiment 3

[0037] A preparation method for bacillus spores, comprising the following steps:

[0038] Step 1) Vegetative cell proliferation:

[0039] Inoculate 1g of Bacillus stearothermophilus into 10g of nutrient-enriched medium and culture it for 12 hours at 52°C with a rotational speed of 200rpm / min; then add 90g of nutrient-enriched medium and continue to cultivate for 12 hours to obtain vegetative cells culture medium;

[0040] Step 2) spore transformation:

[0041] Add 1g of inorganic ionic medium to 1g of vegetative cell culture medium, and culture at 57°C for 12h at a rotational speed of 125rpm / min; then add 1g of inorganic ionic medium every 12h until the added inorganic ionic culture The ratio of the total mass parts of the base to the mass parts of the nutrient-rich medium reaches 8.5:1; a culture solution containing bacillus spores is obtained.

[0042] Among them, the rich nutrient medium preferably includes: 5-20 parts by weight of soybean peptone, 5 parts by weight of b...

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Abstract

The invention relates to a preparation method of bacillus spores. The preparation method comprises the following steps: inoculating 1 part of bacillus stearothermophilus into 10 parts of a nutrient-rich culture medium, and carrying out culturing for 12h at the temperature of 50-55 DEG C and the speed of 150-250 rpm / min; adding with 90 parts of the nutrient-rich culture medium, and carrying out culturing for 12h, thus obtaining nutritive cell culture fluid; adding 1 part of an inorganic ion culture medium to 1 part of the nutritive cell culture fluid, and carrying out culturing for 12h at the temperature of 55-60 DEG C and the speed of 100-150 rpm / min; and adding 1 part of the inorganic ion culture medium every 12h till the ratio of the total parts of the added inorganic ion culture medium to the nutrient-rich culture medium is 7:1 to 10:1, thus obtaining the culture fluid containing the bacillus spores. According to the scheme, only two culture media are needed, the operation is simple, and the repeatability is good; the culture time is shortened, and the consumed time is only 60-72 h; the yield of the spores achieves 10<9> / ml, and the spore rate is 90% or above.

Description

technical field [0001] The invention relates to the field of preparation of microbial spores, in particular to a method for preparing bacillus spores. Background technique [0002] Bacillus stearothermophilus is a Gram-positive bacterium that is aerobic or facultatively anaerobic. Bacillus stearothermophilus can produce spores. The optimum growth temperature is 55-60°C, and the maximum growth temperature is 65-75°C. It is thermophilic bacteria. The collection number of China Industrial Microbiology Culture Collection Center (CICC) is CICC: 10267; or ATCC: 7953. [0003] The spores produced by Bacillus stearothermophilus are non-pathogenic, non-pyrogenic and non-toxic. The spore has definite resistance to certain sterilizing factors, so it can be used as an indicator microorganism to monitor the sterilization effect, including pressure steam sterilization, hydrogen peroxide (H 2 o 2 ) low-temperature plasma sterilization, low-temperature steam formaldehyde sterilization a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12R1/07
CPCC12N3/00
Inventor 尹焕才田晶晶
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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