Method for preparing and regenerating rhizopus oryzae protoplast for producing L-lactic acid at high yield
A technology of protoplast and Rhizoplast oryzae is applied in the field of Rhizoplast oryzae protoplast preparation and regeneration, which can solve the problem of large workload of screening forward mutation, and achieve high protoplast preparation number, high regeneration rate and uniform size. Effect
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[0028] The operation steps of the preparation and the regeneration method of the Rhizopus oryzae protoplast for high production of L-lactic acid are as follows:
[0029] A, preparation Rhizopus oryzae spore suspension:
[0030] The preserved Rhizopus oryzae AS3.819 ( Rhizopus oryzae AS3.819) a loop of mycelia, inoculated onto a solid slope made of 50mL PDA medium in a 100mL Erlenmeyer flask, at 32 o C culture in a constant temperature incubator for 72 hours to obtain a bottle of mycelium containing mature Rhizopus oryzae spores, add 30 mL of sterile distilled water to wash the spores, and obtain a spore liquid containing spore clusters, pour the spore liquid into a bottle containing glass beads In a sterile Erlenmeyer flask, oscillate to break up the spore clusters, filter with 4 layers of sterile high-grade lens-cleaning paper, take the filtrate for multiple dilution, and count under a microscope to make the final concentration of spores 1×10 6 spores / mL, the concentration...
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