A drug-loaded microsphere based on slow and controlled drug release method for preventing and controlling bacteria in bioethanol fermentation

A technology of drug-loaded microspheres and bioethanol is applied in the field of prevention and control of Saccharomyces cerevisiae fermentation and contamination, which can solve the problems of time-consuming and labor-intensive

Active Publication Date: 2021-02-19
BEIJING UNIV OF CHEM TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the time-consuming and laborious shortcomings of the current abuse of antibiotics and the time-consuming and labor-intensive process of fermentation real-time prevention and control, the present invention constructs a drug slow and controlled release method based on pH-responsive poly-4-vinylpyridine microspheres, which can be used for the intelligent prevention and control of miscellaneous bacteria in the fermentation process of Saccharomyces cerevisiae control

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  • A drug-loaded microsphere based on slow and controlled drug release method for preventing and controlling bacteria in bioethanol fermentation
  • A drug-loaded microsphere based on slow and controlled drug release method for preventing and controlling bacteria in bioethanol fermentation
  • A drug-loaded microsphere based on slow and controlled drug release method for preventing and controlling bacteria in bioethanol fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1: Activation of Lactobacillus plantarum

[0058] 1. Activation of Lactobacillus plantarum: Prepare 300mL MRS medium, take three 250mL Erlenmeyer flasks, fill each bottle with 100mL medium, and one bottle labeled "Activated MRS" is used to activate Lactobacillus plantarum ATCC8014. Add 0.75g agar and 1.5g agar respectively to the remaining 2 bottles to make a solid medium, which is reserved for the third step of antimicrobial drug activity determination. Put the aliquoted MRS medium into a high-temperature steam sterilizer, and sterilize it at 115°C for 20 minutes; after the sterilization is completed, take out the medium and cool it in an ultra-clean bench. Lactobacillus plantarum ATCC8014 was inoculated in the MRS medium marked as "activated", and placed in a constant temperature incubator at a constant temperature of 37 ° C for 14 hours of activation culture.

Embodiment 2

[0059] Embodiment 2: yeast activation

[0060] 2. Yeast activation: Prepare 300mL YPD medium, take three 250mL Erlenmeyer flasks, and fill each bottle with 100mL medium, one of which is marked as "activated YPD" for activating Saccharomyces cerevisiae S288c strain, and the other 2 bottles Add 0.75g agar and 1.5g agar respectively to make a solid medium, which is reserved for the third step of antimicrobial drug activity determination. Put the subpackaged YPD medium into a high-temperature steam sterilizer, and sterilize it at 115°C for 20 minutes; after the sterilization is completed, take out the medium and cool it in an ultra-clean bench. Saccharomyces cerevisiae S288c was inoculated in the YPD medium marked as "activated", placed in a constant temperature culture shaker and activated for 20 hours at a constant temperature of 30°C and 150 rpm.

Embodiment 3

[0061] Embodiment 3: antibacterial drug antibacterial activity test

[0062] 3. Antibacterial drug antibacterial activity test: prepare 1 mg / mL of chloramphenicol, penicillin (ready to use), clindamycin and nisin, and use sterile water as a negative control. Pour MRS solid medium containing 1.8% (w / v) agar melted at medium-low heat into the sterilized plate in the ultra-clean bench, wait for it to dry, put a sterilized Oxford cup on it, and put 200 μL of the activated plant The bacteria liquid of Lactobacillus was mixed with 10 mL of MRS solid medium containing 0.75% (w / v) agar melted on medium-low heat and poured into the plate. After the upper culture medium is dry, remove the Oxford cup with sterilized tweezers, add 100 μL of 1 mg / mL chloramphenicol, penicillin, clindamycin, nisin, and sterile water to the removed Oxford cup to form in the small hole. Put the plate into a constant temperature incubator and incubate at a constant temperature of 37°C for 24 hours to observe...

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Abstract

The invention relates to a drug-loaded microsphere-based method for preventing and controlling bacterial contamination in bioethanol fermentation, which belongs to the field of bacterial contamination prevention and control in the field of bioenergy. Prepare 2 mg / mL chloramphenicol solution with citric acid-disodium hydrogen phosphate buffer solution with pH=3, and place poly-4-vinylpyridine microspheres with a particle size of 40-100 μm prepared by suspension polymerization in the chloramphenicol-containing Fully swell in acidic buffer solution, wash with ultrapure water, suction filter, and dry to obtain drug-loaded microspheres; add sterilized drug-loaded microspheres to the fermentation system at an input amount of 2 mg / mL at the beginning of fermentation, Cultivate at 30°C and 150rpm to prevent and control pollution during the fermentation of Saccharomyces cerevisiae. The invention successfully suppresses the reduction of bio-ethanol production caused by bacterial contamination, restores it to the level before bacterial contamination, and improves ethanol production. The invention can reduce the dosage of antibiotics by realizing the pH-responsive release of drugs in the fermentation system, and slow down the problems caused by the abuse of antibiotics.

Description

technical field [0001] The invention relates to a method for preventing and controlling Saccharomyces cerevisiae fermentation contamination, belonging to the field of prevention and control of contamination in the field of bioenergy. Background technique [0002] With the rapid exploitation and depletion of fossil energy, and the increasingly prominent environmental problems such as air pollution, greenhouse effect, and smog, people's demand for renewable and clean energy is becoming increasingly urgent. Bioethanol is favored by experts and scholars from all over the world for its advantages of environmental protection, high energy efficiency and renewable. [0003] At present, researches on the utilization of non-food raw materials of bioethanol, the tolerance of yeast ethanol, and the prevention and control of miscellaneous bacteria pollution are being carried out extensively. Among them, due to unavoidable problems such as instrument leakage and seed contamination, bacte...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/18C12P7/06A01N37/20A01P1/00C12R1/865
CPCA01N37/20C12N1/18C12P7/06Y02E50/10
Inventor 李灏李明陈泽张雅娴
Owner BEIJING UNIV OF CHEM TECH
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