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Method for biosynthesizing poly-beta-hydroxy butyric acid (PHB) by utilizing mixed methane-oxidizing bacteria

A hydroxybutyrate, methane oxidation technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms and other directions, can solve the problems of high PHB extraction cost, inactivation of methane-oxidizing bacteria, low PHB content, etc. The effect of growing cell density, reducing sterilization cost, and easy wall breaking

Inactive Publication Date: 2015-04-15
HARBIN UNIV OF COMMERCE
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  • Summary
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  • Claims
  • Application Information

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Problems solved by technology

[0005] The present invention aims to solve the problems that the low PHB content in the mixed bacteria leads to high extraction cost of PHB, and the impurities in the syngas will lead to the rapid inactivation of methane oxidizing bacteria and cannot grow. Beta-Hydroxybutyrate Method

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  • Method for biosynthesizing poly-beta-hydroxy butyric acid (PHB) by utilizing mixed methane-oxidizing bacteria

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specific Embodiment approach 1

[0013] Specific embodiment 1: In this embodiment, the method for biosynthesizing poly-β-hydroxybutyrate by methane-oxidizing mixed bacteria is carried out according to the following steps:

[0014] The method for biosynthesizing poly-β-hydroxybutyrate by methane-oxidizing mixed bacteria is carried out in the following steps:

[0015] 1. Inoculate the mixed methane oxidizing bacteria liquid into the inorganic salt medium at an inoculation amount of 10%, and cultivate it on a shaker at 32°C and 180 rpm for 5 to 7 days, and replace it with vacuum every 12 hours during this period. fresh gas mixture;

[0016] 2. Then continue to cultivate for 10 to 15 days. During this period, use vacuum method to replace fresh air every 24 hours. After the cultivation, take half of the culture and insert it into fresh inorganic salt medium according to 50% of the inoculum to continue the domestication culture. , repeating this step 10 times to obtain methane oxidizing mixed bacteria that can gro...

specific Embodiment approach 2

[0021] Embodiment 2: This embodiment differs from Embodiment 1 in that: in step 1, the mass percent concentration of the mixed methane-oxidizing bacteria liquid is 30% to 50%. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0022] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the mixed gas in Step 1 and Step 3 is composed of methanation synthesis gas and air in a volume ratio of 1:1. Others are the same as in the first or second embodiment.

[0023] The methanation synthesis gas is a substitute natural gas; biomass is gasified to generate synthesis gas, which is purified and finally methanated to produce substitute natural gas with a methane content > 90%. The synthesis gas contains tar, sulfide, nitride, hydrogen Carbon monoxide, carbon dioxide and other impurities.

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Abstract

The invention relates to a PHB biosynthesizing method, in particular to a method for biosynthesizing poly-beta-hydroxy butyric acid (PHB) by utilizing mixed methane-oxidizing bacteria, and aims to solve the problem of high extraction cost of the PHB caused by low content of the PHB in current mixed bacteria and the problem that methane-oxidizing bacteria are quickly inactivated and cannot grow due to impurities in synthesis gas. The method comprises the following steps: 1, inoculating a bacterium solution of the mixed methane-oxidizing bacteria into an inorganic salt culture medium for culturing, and replacing mixed gas at a time interval of 12h during the period; 2, continuously culturing the bacterium solution, replacing air at a time interval of 24h during the period, putting half of a culture into the inorganic salt culture medium after culturing is finished, and continuously performing habituated culture for 10 times to obtain the mixed methane-oxidizing bacteria which are capable of growing in the mixed gas and are rich in the PHB; 3, inoculating the mixed methane-oxidizing bacteria which are obtained by screening into the inorganic salt culture medium, replacing the air with the mixed gas, performing culturing, ending culturing, and determining the content of the PHB. The method is used for synthesizing the PHB.

Description

technical field [0001] The invention relates to a method for biosynthesizing poly-β-hydroxybutyrate. Background technique [0002] Poly-β-hydroxybutyrate (PHB) exists in most microbial systems as an energy reserve, and has good biodegradability and biocompatibility. Using PHB to make various packaging materials can greatly reduce the pollution of waste to the environment , fundamentally solve the "white pollution" problem. At present, microbial fermentation production is the main means of synthesizing PHB. However, due to the high cost of fermentation, it is difficult to open up a broad application market for PHB production. Methane-oxidizing mixed bacteria is a combination of various symbiotic bacteria with methane-oxidizing bacteria as the main body. Compared with pure bacteria, it is easier to operate, less susceptible to contamination by miscellaneous bacteria, and has high stability. It is more conducive to the continuous industrial production of biosynthetic PHB. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P39/00C12P7/62C12R1/01
Inventor 董静辛嘉英
Owner HARBIN UNIV OF COMMERCE
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