A method for preparing bio-enzyme preparation, wherein comprises the following steps:
- (1) selecting a yeast and lactic acid bacteria combination as a culture, the yeast and lactic acid bacteria combination comprises Candida ethanolica B-JJ1, Lentilactobacillus buchneri B-JR1, Lactobacillus paracasei B-JR2, Lactobacillus zeae B-JR4, Lactobacillus plantarum B-JR5 and Lactobacillus chiayiensis B-JR6;
- the Candida ethanolica B-JJ1 with the deposit number:
- CCTCC NO:
- M2021136;
- the Lentilactobacillus buchneri B-JR1 with the deposit number:
- CCTCC NO:
- M2021132;
- the Lactobacillus paracasei B-JR2 with the deposit number:
- CCTCC NO:
- M2021133;
- the Lactobacillus zeae B-JR4 with the deposit number:
- CCTCC NO:
- M2021135;
- the Lactobacillus plantarum B-JR5 with the deposit number:
- CCTCC NO:
- M2021501;
- the Lactobacillus chiayiensis B-JR6 with the deposit number:
- CCTCC NO:
- M2021502;
- (2) preparation and inoculation of a medium of the culture:
- the medium is prepared with mashed potato powder, wheat germ and glutinous rice as main ingredients;
- the medium is steamed after adding water and stirring evenly, and the preparation of the medium has been completed when temperature of the medium is cooled to below 45° C. in aseptic conditions;
- homogeneously mixing the prepared medium with the culture;
- (3) adjusting humidity of the culture medium after inoculation in step (2) to ensure that there are water seeps on the medium and the whole medium is kept moist and breathable;
- (4) processing aerobic fermentation of the medium in step (3) until the medium uniformly appears white and milky white bacterial plaque to be visible with the naked eye, and then transferring the medium into a fermenter;
- to close all channels of the fermenter to enable the medium fermentations into the fermenter to continue aerobic culture in the fermenter until the oxygen in the fermenter is depleted and enters anaerobic fermentation state;
- (5) when there is no pressure change in the fermenter, solid-state fermentation enters into saturation fermentation, and there is a strong fermentation fragrance when opening the tank, with abundant mycelium covering the surface of the solid medium to be visible with the naked eye;
- taking the fermentation products for testing, and the total number of viable yeast is 6.4×104-5.9×105 cfu/ml;
- the total number of viable lactic acid bacteria is 4×108-5.1×109 cfu/ml;
- and at this time the solid fermentation process has reached saturation point, and then enters the liquid fermentation stage;
- (6) adding the solid-state fermentation in the step (5) to sterile water and stirring thoroughly, and carrying out aerobic activation and cultivation until the surface of the liquid ferment is densely covered with white or milky white bacterial plaque;
- to close all channels of the fermenter and again allow the fermentation to gradually deplete oxygen and enter into the anaerobic fermentation process;
- (7) when the bacterial plaque on the surface of the liquid fermentation in step (6) is in folding state, the middle and upper layers of clear liquid appear golden yellow, clear and transparent, and the solid fermentation to liquid fermentation process is regarded as full complement, when the bottom layer of solid fermentation and the middle layer of liquid are clearly layered and not sticky, and the fermentation product obtained is the bio-enzyme preparation.