Moldfungi suspension culture method by high throughput screening

Abstract

The invention relates to a fungi culture method for the technical field of bioengineering and biology to realize high throughput screening on strains, in particular to a moldfungi suspension culture method by high throughput screening. The method comprises the steps of selecting moldfungi, obtaining bacterial colony, setting a perforated plate culture environment, suspension culturing and the like. Through a simple mode to ensure that the bacterial colony is suspended on the surface of a liquid medium for static culturing, solves the oxygen supply problem of miniature culture, realizes high throughput culturing of the moldfungi, is associated with corresponding high throughput detecting technology to realize high throughput screening, ensures that the culturing process of the strains and the forming process of spore are combined into a whole, and further shortens the flow of strain breeding to achieve the aims of high yield strains and high throughput screening. The method has simple operation and high efficiency.

Description

technical field [0001] The invention relates to a strain culture method used in the fields of bioengineering and biotechnology to realize high-throughput screening of strains, in particular to a suspension culture method for high-throughput screening of filamentous bacteria. Background technique [0002] At present, microorganisms in nature except viruses are mainly composed of filamentous bacteria and non-filamentous bacteria, non-filamentous bacteria include bacteria and yeasts, filamentous bacteria include: actinomycetes and molds, the main characteristics of filamentous bacteria It is filamentous (for example, common moldy phenomena are caused by molds), the hyphae of actinomycetes are very thin, and they are prokaryotic organisms, and the hyphae of molds are thicker, with a diameter about 10 times that of actinomycetes, which is eukaryotic biology. Many important products of bioengineering are produced by filamentous microorganisms (including molds and actinomycetes), ...

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Problems solved by technology

However, the strain culture technology of the high-throughput screening platform develops relatively slowly. The main reason is that high-throughput culture means miniaturized culture, and the miniaturized culture mode is limited by the existing culture equipment, especially the current useful filamentous bacteria are all It is an aerobic bacteria, and the oxygen supply mode has become the bottleneck of miniaturized culture. Some foreign companies have developed micro-reactor equipment containing multiple micro-sensor probes, which can meet the supply of oxygen in the liquid micro-cultivation process and the related conditions in the culture process. Detection and control (temperature, pH, optical density, etc.) The main mode of culture is still liquid uniform culture, that is, micro-fermentation tanks, some products have been used, but the cost is expensive

[0005] In China, the evaluation system for strain screening of filamentous bacteria is based on liquid shake flask culture as a conventional strain culture technology. The operation process is complicated, the equipment efficiency is low, the culture scale is small, and there is no good detection technology for high-throughput screening. , thus greatly limiting the screening efficiency

At present, a large-scale liquid shake flask culture equipment can cultivate up to 100 strains of bacteria per day, and the cost of the equipment is 30,000 to 40,000 yuan, and the probability of obtaining good strains is about 10. -4 -10 -6 Between, that is to say, it is possible to obtain an excellent strain after working continuously for 100-10000 days, the workload is heavy, and the screening efficiency is low

And liquid shake flask culture, in order to obtain oxygen supply, shake flask needs to be shaken ceaselessly, in shaking process, can interrupt the hyphae of filamentous fungus, can't produce asexual spore (second-generation propagule) in the culture process , further extending the process of strain selection

[0006] In the prior art, filamentous bacteria can also grow on solid medium, and can also produce metabolites, which can be used for preliminary qualitative screening of strains, but the product detection is inconvenient, and the absorption rate of nutrients is affected, which will affect the screening As a result, so the real screening test has to be converted to liquid shake flask culture

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