Method for improving cadmium tolerance of bacillus through mutagenesis

A Bacillus, tolerance technology, applied in the field of improving the tolerance of microorganisms to cadmium, to achieve the effect of improving tolerance

Inactive Publication Date: 2011-01-26
NANKAI UNIV
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Problems solved by technology

At home and abroad, the ultraviolet mutagenesis breeding technology is mainly used to obtain a large number of high-yield industrial mi

Method used

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Examples

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Embodiment 1

[0018] UV mutagenesis and first screening: the number of colonies was 1×10 6 Inoculate 1mL of Bacillus subtilis bacilli per mL into 100mL liquid beef extract peptone, put it into a 250mL Erlenmeyer flask with a silica gel stopper, and culture with shaking at 200r / min for 10h. The culture medium was removed by centrifugation, and the bacterial suspension was prepared with physiological saline. Add glass balls to vibrate to disperse, and filter with absorbent cotton to form single cells, and the degree of dispersion reaches 90-95%. The prepared bacterial suspension 3mL (1 × 10 6 per mL) were placed in a flat-bottomed petri dish with a diameter of 7 cm, and placed on a magnetic stirrer at 37°C for ultraviolet mutagenesis. Before mutagenesis begins, the 15W power UV lamp must be preheated for 20 minutes to stabilize the light wave (254nm). Adjust the distance between the culture dish and the lamp tube to be 30 cm, open the lid of the culture dish, and carry out mutagenesis in a...

example 2

[0021] Count the colonies to 1 x 10 6 Each / mL cadmium-tolerant mutant B38 and the starting bacteria were inoculated in 100 mL of beef extract peptone medium containing cadmium 3mmol / L. After shaking at 200r / min for 10 hours, the B38 mutant bacteria could grow, but the starting bacteria could not grow. .

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Abstract

The invention relates to a microorganism breeding method, in particular to a method for performing eukaryon and pronucleus microorganism protoplasm multi-parent fusion by establishing genome-based reorganization technology. The technical scheme adopted to fulfill the aim of the invention comprises the following steps of: performing induced mutation on a microorganism and screening so as to obtain a mutated microorganism having high cadmium resistance; and performing slant spore culturing, strain mutagenesis, genome reorganization, screening and multiple recursion infusion by taking the mutated microorganism with the high cadmium resistance as an initial strain by a cell protoplast multi-parent fusion method so as to obtain an anti-cadmium strain with good property. When the cell protoplast multi-parent fusion method established by the invention is used for transforming bacillus subtillis and candida tropicalis which are resistant to cadmium, two high-quality infusion strains which have extremely higher cadmium resistance than that of the bacillus subtillis and the candida tropicalis are obtained.

Description

technical field [0001] The invention relates to a method for improving the tolerance of microorganisms to cadmium by means of microorganism mutagenesis. Background technique [0002] Cadmium (Cd) is a heavy metal element with strong biological toxicity. Together with Pb, Cr, As and Hg, it is called "five poisons", ranking second. In recent years, with the development of modern industry and the wide application of cadmium in smelting, chemical industry, papermaking and electronic industries, etc., the pollution of Cd has become increasingly serious. Therefore, how to remediate cadmium pollution in the environment has become a hot research topic at home and abroad. At present, the research on microbial remediation of cadmium pollution mainly focuses on the selection strategy of cadmium-tolerant microorganisms. The traditional screening of cadmium-tolerant microorganisms mainly adopts the method of "domestication". Many studies have shown that microorganisms that have been "d...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N13/00C12Q1/04C12R1/125
Inventor 姜春晓孙红文张彦峰王婷张清敏
Owner NANKAI UNIV
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