Chlorella zofingiensis heterotrophic high-density culture method

Abstract

The invention discloses a Chlorella zofingiensis heterotrophic high-density culture method which is characterized in that Chlorella zofingiensis is inoculated into a fermentation tank and cultured, aused basal culture medium and a used fed-batch culture medium take ammonium salt as a nitrogen source, the carbon nitrogen ratio of the basal culture medium is 5:(1-80):1, the carbon nitrogen ratio ofthe fed-batch culture medium is 5-20 times that of the basal culture medium, the pH (potential of hydrogen) value of a culture system is monitored in real time during culture, adjusted by adding ammonia water according to monitoring results and kept 6.5+ / -0.2 to supplement the nitrogen source, the glucose concentration of the culture system is monitored in real time during culture, feeding of thefed-batch culture medium is started when the glucose concentration of the basal culture medium is reduced to 3-4 g / L, and the variation amplitude of the glucose concentration of the culture system islower than 20% within any two hours by the aid of feeding speed. According to the characteristic of pH reduction in the ammonium salt using process of cells, the pH is regulated by the aid of the ammonia water to supplement the nitrogen source. The culture method can effectively avoid the inhibitory effect of nitrogen source excess or deficiency in an existing fixed carbon nitrogen ratio feedingculture mode on cell growth.

Description

technical field [0001] The invention relates to the technical field of microalgae cultivation, in particular to a method for heterotrophic high-density cultivation of chlorella. Background technique [0002] Astaxanthin is a purple-red ketone carotenoid with strong coloring and anti-oxidation functions, and is widely used in aquaculture, food, medicine, cosmetics and other fields. Chlorella Chlorella zofingiensis It is a single-cell green algae, which grows fast and can efficiently accumulate astaxanthin. It is considered to be another source of natural astaxanthin production that has attracted much attention after Haematococcus pluvialis and Phaffia rhodozyme [Document 1: IP PF, Chen F. Peroxynitrite and nitrylchloride enhance astaxanthin production by the green microalga Chlorellazofingiensis in heterotrophic culture. Process Biochemistry, 2005, 40: 3595-3599]. Chlorella sp. as a novel potential microalgae for industrial astaxanthin production Chlorella zofingiensis Not...

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Problems solved by technology

[0007] In order to solve the existing chlorella Chlorella zofingiensis The problem of slow cell proliferation and low biomass concentration when using nitrate nitrogen and fixed carbon-nitrogen ratio in the high-density culture process, the invention provides a kind of chlorella Chlorella Zofingiensis The heterotrophic high-density culture method uses ammonium salts with low energy consumption in the process of absorption and utilization as the nitrogen source, and at the same time abandons the traditional fixed carbon-nitrogen ratio feeding, instead, it is based on the pH drop caused by Chlorella in the process of using nitrogen sources. Adaptively supplement ammonia water, during the process of supplementing ammonia water, on the one hand, maintain an appropriate and constant pH in the heterotrophic environment of microalgae by supplementing ammonia water, on the other hand, ammonia water is also used as a nitrogen source supplement for algae cell growth Therefore, the nitrogen source provided by ammonia water is actually supplemented adaptively according to the nitrogen utilization and demand of microalgae cells, thereby avoiding the limitation of microalgae growth caused by feeding with a fixed carbon-nitrogen ratio, and improving the growth rate of cells. Growth Rate and Biomass Concentration Purposes for Chlorella Chlorella zofingiensis Provide technical support for heterotrophic high-density culture of its mutants

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