Rhizopus oryzae non-carrier immobilization cell culturing method

Abstract

The invention discloses a cell cultivation method of immobilized rhizopus oryzae without a carrier. The steps are as that: (1) spores are inoculated with high concentration and then thick spores-suspended emulsion is formed. (2). a number of bacterium balls are produced by controlling the aeration condition. The proportion of sodium ion and potassium ion in the culture medium is controlled to leadthe rhizopus oryzae spawn to increase and keep the stability of the cell form. Methyl silicone oil is added to form a plurality of bacterium ball cells with certain diameter. The bacterium balls cutflow and the immobilized rhizopus oryzae without a carrier are obtained. (3) when the bacterium ball obtained is transferred to a big pot, the upgrade of the bacterium balls and the increasing of thenumber are controlled. The invention has the advantages that firstly, the appearance of bacterium ball formed in the suspending culture system is regular and the size is equal; secondly, the conditionof bacterium ball accumulated by cells is mild and controllable; thirdly, the density of bacterium ball cells is more than 104 / L and the cell activity is more than 95 percent; fourthly, the large-scaled culture is easy to realize; fifthly, the cost of immobilized carrier can be saved.

Description

technical field [0001] The invention relates to the technical field of microbes and cell engineering, specifically utilizes the tendency of rhizopus oryzae cells to aggregate and flocculate to form hyphal balls in a stirred culture system, and implements carrier-free immobilization of rhizopus oryzae without using a cell immobilization medium method of cell culture. Background technique [0002] As one of the important organic acids, lactic acid is widely used in food, medicine, feed, chemical industry and other fields. The use of L-lactic acid in the food industry is absolutely safe, so the World Health Organization advocates the use of L-lactic acid in the food and pharmaceutical industries to replace the currently widely used DL-lactic acid. The polymer of L-lactic acid—poly-L-lactic acid (PLA) is a non-toxic polymer compound with biocompatibility. Poly-L-lactic acid is a good green material and is of great significance to eliminate "white pollution". Therefore, the bio...

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Problems solved by technology

The use of carrier materials has brought many problems in cell physiology, ecology, technology, engineering and economy, which are mainly reflected in: (1) In terms of physiology, carrier materials sometimes have relatively large physiological toxicity to cells, affecting the target Biosynthesis of metabolites; (2) In terms of ecology, carrier materials separate cells or small cell groups from each other, which is easy to form an environment that is not conducive to the coordination between cells and affects the biosynthesis of metabolites; (3) In terms of technology, The large-scale preparation process of carrier-immobilized cells is complicated, and it is easy to cause contamination by bacteria; (4) In terms of engineering, on the one hand, the mass transfer performance of carrier-immobilized cells is poor, and the concentration gradient of the external nutrient matrix and internal metabolites is large, resulting in Its overall activity is low. On the other hand, the flushing of the fluid flow in the bioreactor and the expansion of the gas generated in the metabolic process will also cause the rupture of the immobilized cells of the carrier, affecting the life of its continuous use; (5) In terms of economy, The use and constant consumption of carrier materials brings additional costs, making immobilized cell technology difficult to promote and apply in the traditional fermentation industry

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