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Artificial flora cellulose biotransformation method

A technology of cellulose and composite flora, applied in the biological field, can solve problems such as low conversion efficiency, hidden dangers of biological safety, and strict requirements for sterility in the process

Active Publication Date: 2019-04-23
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the deepening of research, whether it is super engineered bacteria or natural complex flora, there are inherent shortcomings
Super engineering bacteria have problems such as limited ability to express foreign proteins, poor genetic stability, difficulty in genetic modification, narrow substrate range, low degradation efficiency, strict sterility requirements in the process, potential biological safety hazards, and fermentation susceptible to product inhibition; natural flora There are many problems such as low fermentation efficiency, poor system stability, low controllability, low conversion efficiency, and biological safety hazards.

Method used

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  • Artificial flora cellulose biotransformation method
  • Artificial flora cellulose biotransformation method
  • Artificial flora cellulose biotransformation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Embodiment 1, the preparation of composite flora

[0077] During the long-term selection of natural flora in nature, a harmonious symbiotic relationship has been formed, and through the complementation of metabolic pathways, a more efficient synergistic metabolic system for cellulose conversion has been established. Through the research and understanding of these symbiotic relationships and synergistic metabolic mechanisms, it is easier to establish a simplified flora system with stable flora structure and better fermentation performance. The inventors of the present invention have gone through a lot of experiments, based on the research on the mechanism of efficient synergistic conversion of cellulose between strains and the molecular mechanism of strain symbiosis in the natural flora that converts cellulose into biofuels reported in the literature, screened the strains and carried out artificial recombination, and obtained A complex bacterial flora that can efficientl...

Embodiment 2

[0100] Embodiment 2, the stability detection of composite flora

[0101] 1. Stability detection of complex flora A

[0102] 1. Inoculate the composite flora A prepared in Example 1 into 90 mL of restricted medium 1 to obtain a mixed solution; in the mixed solution, the content of Clostridium perfringens is 4 × 10 5 cfu / mL, the content of Clostridium thermocellum was 16×10 5 cfu / mL, the content of thermotolerant Clostridium is 42×10 5 cfu / mL, the content of Bacillus thermosphaeroids is 29×10 5 cfu / mL, the content of Pseudoxanthomonas in Taiwan, China is 9×10 5 cfu / mL.

[0103] 2. Take the mixed solution obtained in step 1, and culture it at 55° C. for 5 days to obtain the first-generation fermentation solution. Inoculate 10 mL of the first-generation fermentation broth into 90 mL of restricted medium 1 and culture at 55°C for 5 days to obtain the second-generation fermentation broth. Inoculate 10 mL of the second-generation fermentation broth into 90 mL of restricted medi...

Embodiment 3

[0136] Embodiment 3, the mensuration of the total output of organic product produced by complex flora fermentation

[0137] 1. Inoculate the composite flora A prepared in Example 1 into 90mL restrictive medium 1 to obtain a mixed solution; in the mixed solution, the total content of bacteria is 1×10 7 cfu / mL, the content of Clostridium perfringens is 4×10 5 cfu / mL, the content of Clostridium thermocellum was 16×10 5 cfu / mL, the content of thermotolerant Clostridium is 42×10 5 cfu / mL, the content of Bacillus thermospheroidus is 29×10 5 cfu / mL, the content of Pseudoxanthomonas in Taiwan, China is 9×10 5 cfu / mL. The mixture was cultured at 55° C. for 3 d or 5 d to obtain a fermented liquid.

[0138] 2. Inoculate Clostridium thermocellum into 90 mL of restricted medium 1 to obtain a mixture; in the mixture, the content of Clostridium thermocellum is 1×10 7 cfu / mL. The mixture was cultured at 55° C. for 3 d or 5 d to obtain a fermented liquid.

[0139] 3. Inoculate Clostrid...

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Abstract

The invention discloses an artificial flora cellulose biotransformation method. An artificial flora is composed of Clostridium, Ureibacillus and Pseudoxanthomonas, and the colony formation unit numberratio is (3-70):(10-50):9. Organic products comprise ethanol, acetic acid, acetone, butyric acid, butanol and the like and can serve as biofuel or chemical raw materials. Experiments prove that the artificial flora conversion rate, the raw material conversion rate, the cellulose degradation capacity and the organic product total output are significantly increased, and the raw material conversionrate reaches up to 0.473 g / g substrate. The method is easy to operate, the cost is low, and the method has popularization and application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for converting cellulose raw materials with artificial flora to produce organic products. The organic products include ethanol, acetic acid, acetone, butyric acid and butanol, which can be used as biofuels or chemical raw materials. Background technique [0002] Cellulose is the polysaccharide with the highest stock and yield in nature, mostly in the form of agricultural and forestry waste. Cellulose resource utilization is a current research hotspot, and the conversion of cellulose into small organic molecules such as biofuels is an important way to obtain renewable fuels and chemical raw materials. Biotransformation technology has the advantages of mild conditions, less process pollution, and low energy consumption, and is considered to be one of the most potential conversion methods. In the biotransformation technology, one-step biological conversion of cellu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/06C12P7/54C12P7/28C12P7/52C12P7/16C12R1/145C12R1/01
CPCC12N1/20C12P7/06C12P7/16C12P7/28C12P7/52C12P7/54Y02E50/10
Inventor 闫建斌杜然郭律均李十中谢道昕
Owner TSINGHUA UNIV
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