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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.

Active Publication Date: 2021-06-04
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Genetic modification of microorganisms to regulate the metabolism of guaiacols can strengthen its production pathway from the source, which is currently the most effective method. create uncontrollable effects
Although guaiacols can be produced, the production is rarely enough to enhance the taste of soy sauce

Method used

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  • Weissella for promoting zygosaccharomyces rouxii to produce guaiacol substances
  • Weissella for promoting zygosaccharomyces rouxii to produce guaiacol substances
  • Weissella for promoting zygosaccharomyces rouxii to produce guaiacol substances

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses Weissella for promoting zygosaccharomyces rouxii to produce guaiacol substances, and belongs to the technical field of biological engineering. By co-culturing Weissella paramesenteroides LCW-28 and zygosaccharomyces rouxii ZQ-01 in a culture medium, the yield of guaiacol substances is effectively increased; the total yield of the guaiacol substances is the highest; the total yield of the guaiacol substances is increased by 3.87 times compared with the total yield of the guaiacol substances cultured independently, is increased by 15.71 times compared with candida, and is increased by 17.28 times compared with torulopsis glabrata, wherein the yield of 4-ethyl guaiacol is 2.12 mg / L, and is 8.8 times of that of the single culture of the zygosaccharomyces rouxii.

Description

technical field [0001] The invention relates to a strain of Weissia bacterium that promotes the production of guaiacol by Zygomyces ruckeri, and belongs to the technical field of bioengineering. Background technique [0002] The guaiacols include guaiacol, 4-ethyl guaiacol, and 4-vinyl guaiacol, which have smoky, soy sauce and mild barbecue flavors, which are recognized as characteristic flavors of soy sauce One of the substances, it is a seasoning sauce and an important natural essence. Appropriate content of guaiacol can enhance the richness of soy sauce and alleviate the salty taste of soy sauce. If there is little or no guaiacol, the taste of soy sauce will be weak, and the content difference of 0.5mg / L will be easily recognized by the senses. Guaiacol is one of the main flavor components of soy sauce, so it is necessary to increase its content in soy sauce to an appropriate concentration. [0003] At present, the guaiacols in soy sauce are mainly increased from two a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16C12N1/04C12G3/02C12G1/00C12C11/02C12C11/00C12J1/00A23L27/50C12R1/01C12R1/645
CPCC12N1/20C12N1/16C12N1/04C12G3/02C12G1/00C12C11/02C12C11/00C12J1/00A23L27/50
Inventor 方芳胡光耀堵国成陈坚
Owner JIANGNAN UNIV
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