Rhizopus oryzae non-carrier immobilization cell culturing method

A carrier-free immobilization and cell culture technology, applied in the field of microbiology and cell engineering, can solve the problems of unfavorable cell coordination, affecting lifespan, and low overall activity, and achieve the effects of smooth synthesis, easy repeated use, and high fermentation intensity

A carrier-free immobilization and cell culture technology, applied in the field of microbiology and cell engineering, can solve the problems of unfavorable cell coordination, affecting lifespan, and low overall activity, and achieve the effects of smooth synthesis, easy repeated use, and high fermentation intensity

CN100590191CInactive Publication Date: 2010-02-17HEFEI UNIV OF TECH
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  • Rhizopus oryzae non-carrier immobilization cell culturing method

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Experimental program
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Embodiment

[0026] Rhizopus oryzae carrier-free immobilized cell culture steps are as follows:

[0027] (1) Preparation of Rhizopus oryzae spores

[0028] The rhizopus oryzae strain Rhizopus oryzae (Rhizopus oryzae / AS 3.819) was streak-inoculated on the potato dextrose (PDA) plate, and the culture temperature was set at 32 ° C on the first day, and the relative culture humidity was 70%, which was conducive to mycelial growth. It is 34 ℃ to set the culture temperature every day, and the relative culture humidity is 30%, which is beneficial to the generation of spores. After the black dense spores completely cover the white hyphae, they can be ready to collect, and mature Rhizopus oryzae spores are obtained.

[0029] (2) Preparation of Rhizopus oryzae spore emulsion suspension

[0030] All the Rhizopus oryzae spores in the 250mL Erlenmeyer flask are all scraped and washed into sterile water, and then inoculated in a 5L fermentation tank with a straight plate stirring paddle. The medium in ...

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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...

Claims

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Application Information

Patent Timeline
17 Feb 2010
Publication
CN100590191C
IPC
C12N1/14; C12N11/00
Inventors
姜绍通; 潘丽军