Weissella for promoting zygosaccharomyces rouxii to produce guaiacol substances
A technology of Zygomyces rouckeri and Weissella, applied in the field of bioengineering, can solve the problems of uncontrollable influence on the safety of soy sauce flavor, low yield and insufficient to improve the taste of soy sauce, etc.
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Embodiment 1
[0036] Example 1: Effect of co-cultivation on the growth of Weissella enteroides and Zygomyces ruckeri
[0037] Weissella enteroides LCW-28 was statically cultured in MRS at 37°C for 24 hours to obtain a pre-culture. The preculture was inoculated into 100 mL of fresh MRS medium at an inoculum size of 2% (v / v). After static incubation for 24 h, the cells were collected by centrifugation at 10,000 g for 5 min, and resuspended in 4 mL sterile water. Transfer 1 mL of the suspension to a 50 mL centrifuge tube filled with 30 mL of MRS medium and grow statically at 37 °C for 4 h.
[0038] Zygomyces rouckeri was grown statically at 30° C. for 24 h in YPD medium containing 6% (m / v) NaCl to obtain a pre-culture. The preculture was inoculated into 100 mL of fresh YPD broth containing 6% (m / v) NaCl at an inoculum size of 2% (v / v). After static incubation for 24 h, the cells were collected by centrifugation at 10,000 g for 5 min, and resuspended in 4 mL sterile water.
[0039] For pure...
Embodiment 2
[0044] Example 2: The effect of co-cultivation of different microorganisms on the yield of guaiacols
[0045] ZQ-01, Candida orthopsilosis, or Torulopsis glabrata were mixed with Weisseria enteroidea LCW-28, Staphylococcus pasteuri in MRS medium, respectively. ), Tetragenococcus halophilus, Bacillus cereus or Pediococcus acidilactici were co-cultured.
[0046] In a 100 mL culture flask containing 30 mL of MRS medium, the co-cultured bacterial solution (3.4×10 7 CFU / mL), cultured at 37°C for 60h, and sucked the fermentation broth every 10h. The amount of guaiacol in the fermentation supernatant was detected.
[0047] The fermentation broth was centrifuged at 10,000 r / min for 5 min at 4 °C, the supernatant was collected, and 5 times the amount of dichloromethane and 83.36 μg·L were added. -1 The 2-octanol was used as the internal standard for ultrasonic extraction for 15 min, and the organic phase was obtained through a separatory funnel, and the extraction was repeated twice...
Embodiment 3
[0051] Example 3: The effect of co-culture on the expression of important genes on the synthetic 4-EG pathway
[0052] Centrifuge the cultured L. brucei suspension at 12,000 r / min for 3 min at 4°C, discard the supernatant, place it in a centrifuge tube (DEPC water treatment) and put it in liquid nitrogen. Grind in a cold mortar, grind the cells to a white powder, and transfer to a 1.5 mL Eppendorf tube. RNA was extracted using Nasia kit, and the reverse transcribed cDNA was extracted using THUNDERBIRD The qPCR Mix kit was used for the qRT-PCR reaction according to the instructions of the instrument. The primers used are shown in (Table 2).
[0053] When co-cultured for 4 hours, the relative expression of genes differed the most, among which the relative expression of aroB and aroF increased the most, which increased by 4.5 times and 3.2 times compared with the control group ( image 3 ). However, pheA, the main gene for synthesizing phenylalanine, was down, which proved t...
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