High-throughput screening method of high-yield monascus pigment strains

Abstract

The invention discloses a high-throughput screening method of high-yield monascus pigment strains and belongs to the technical field of high-throughput screening. According to the method, a solid-state fermentation mode is utilized to represent the yield of liquid-state fermentation pigments, so that the problems that the dissolved oxygen content during liquid-state fermentation of pore plates islow and an extremely low color value or even fermentation failure is resulted from the fact that mycelia are likely to grow into spheres during fermentation are solved successfully. In combination with an ARTP mutagenesis technology and a flow-cytometry efficient sorting technology, high-throughput rapid screening of monascus with high-yield monascus pigments is successfully achieved, a shaking flask verification effect is excellent, manpower and material resources are greatly saved, the screening efficiency is improved, and an effective method is provided for high-throughput screening of monascus.

Description

Technical field [0001] The invention relates to a method for high-throughput screening of monascus pigment high-yielding strains, and belongs to the technical field of high-throughput screening. Background technique [0002] Monascus red is a natural pigment produced by Monascus with high medicinal and nutritional value. Monascus red pigment is widely used as a toner in the fields of meat products, pasta products, beverages and medicine, and is favored by consumers because of its safety and harmlessness. [0003] Breeding high-yielding strains of monascus pigment is a direct and effective strategy to increase fermentation yield. Compared with traditional physical and chemical mutagenesis and plate separation and screening, ARTP mutagenesis has the advantages of high efficiency, safety, and convenience, and it can save a lot of manpower and material resources through flow cytometry sorting, high-throughput culture and screening. At the same time, the screening cycle is shortened, ...

AI Technical Summary

Problems solved by technology

[0004] Currently commonly used mutagenesis methods include physical mutagenesis and chemical mutagenesis. Ultraviolet mutagenesis has the advantages of long use time, good effect, and simple equipment. However, the energy of ultraviolet mutagenesis is relatively low, and it can only cause a single damage and mutation

The mutation rate of chemical mutagenesis is usually higher than that of physical mutagenesis, and it is very economical, but most of the mutations it causes are base swaps, which are prone to reverse mutations, resulting in unstable mutations; in addition, chemical mutagenesis uses Most of the chemical mutagens are strong carcinogens and have high potential safety hazards

[0005] Numerous studies have shown that excellent strains with high production of physiologically active products can be obtained by mutation breeding, but there are shortcomings such as randomness and blindness, cumbersome workload, and unpredictable results in the screening process

When the current classic mutagenesis combined with high-throughput screening method is applied to screen high-yield strains of Monascus, the dissolved oxygen in the orifice plate is low, and Monascus is easy to grow into a ball during liquid fermentation in the orifice, resulting in color and price. As low as a few units or fermentation failure, the mutagenic effect cannot be clearly reflected, reducing the effectiveness of screening

In addition, the fermentation period of Monascus is relatively long, too much evaporation will cause the actual value of the color price ratio to be high, and it will also affect the normal growth of mycelium, and even the medium will dry out, resulting in the failure of liquid fermentation

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