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A Weissella strain that promotes the production of guaiacols by Zygomyces rouckeri

A technology of Zygosaccharomyces rutheli and Weisseria, which is applied in the field of bioengineering, can solve the problems of insufficient output to improve the taste of soy sauce, uncontrollable influence of safety soy sauce flavor, and the like

Active Publication Date: 2022-05-10
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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  • A Weissella strain that promotes the production of guaiacols by Zygomyces rouckeri
  • A Weissella strain that promotes the production of guaiacols by Zygomyces rouckeri
  • A Weissella strain that promotes the production of guaiacols by Zygomyces rouckeri

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: Effects of co-cultivation of different microorganisms on the yield of guaiacols

[0045] Zygomyces ruckeri ZQ-01, Candida orthopsilosis, or Torulopsis glabrata were mixed with Weissella enterica-like LCW-28, Staphylococcus pasteuri ), Tetragenococcus halophilus, Bacillus cereus or Pediococcus acidilactici were co-cultured.

[0046] In a 100mL culture bottle containing 30mL MRS medium, add the co-cultivated bacterial solution (3.4×10 7 CFU / mL), cultivated at 37°C for 60 hours, and sucked the fermentation broth every 10 hours. The amount of guaiacols in the fermentation supernatant was detected.

[0047] Centrifuge the fermentation broth at 10000r / min for 5min at 4°C, collect the supernatant, add 5 times the amount of dichloromethane and 83.36μg·L -1 2-octanol was used as an internal standard for ultrasonic extraction for 15 min, and the organic phase was obtained through a separatory funnel, and the extraction was repeated twice. Add 5 g of anhydrous Na2SO...

Embodiment 3

[0051] Embodiment 3: Co-cultivation influences the expression of important genes on the synthetic 4-EG pathway

[0052] Centrifuge the cultured Saccharomyces ruxii suspension at 12000r / min for 3min at 4°C, discard the supernatant, put it in a centrifuge tube (treated with DEPC water) and put it in liquid nitrogen, and pour it into the pre- Grinding in a cold mortar, the bacteria were ground into a white powder, and transferred to a 1.5mL Eppendorf tube. Nasia kit was used for RNA extraction, and the reverse transcribed cDNA was used for THUNDERBIRD qPCR Mix Kit Follow the instructions of the instrument for qRT-PCR reactions. The primers used are shown in (Table 2).

[0053] When co-cultured for 4 h, the relative expression of genes had the largest difference, and 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 the synthesis of phenylalani...

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Abstract

The invention discloses a strain of Weissia bacterium that promotes the production of guaiacol by Zygomyces ruckeri, belonging to the technical field of bioengineering. The production of guaiacols was effectively improved by co-cultivating Weissella enteritidis LCW‑28 and Zygosaccharomyces rouckeri ZQ‑01 in the medium, and the total production of guaiacols was the highest compared with the single culture Increased by 3.87 times, compared with Candida, increased by 15.71 times, and compared with Toruula glabrata, increased by 17.28 times; wherein, the 4-ethyl guaiacol output was 2.12mg / L, which is the only method for culturing Lushi 8.8 times that of Zygomyces.

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 Patents(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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