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Method for preparing hydrogen by fermentation through using special anaerobic clostridium pasteurianum

A technology of anaerobic clostridium pasteurii and anaerobic bacteria is applied in the field of hydrogen production using efficient anaerobic bacteria, which can solve the problems of anaerobic fermentation conditions, difficult to popularize, complicated operation and the like

Active Publication Date: 2011-03-23
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Through patent search, the existing patents are mainly research on fermentation substrates, fermentation equipment and improving hydrogen production efficiency, such as a kind of sludge and organic waste mixed biological production Hydrogen method (application number: 200710032658.0 Zhou Shaoqi, etc.), photosynthetic microbial hydrogen production reactor (application number: 200620032282.4), biological fermentation hydrogen production device (application number: 00231651.X Shen Jianquan), a CO2 sub>The method of rapidly multiplying algae and directly using it for biological hydrogen production (200710010804.X) and the expanded bed equipment for high-efficiency fermentation biological hydrogen production (application number: 03260012.7, Ren Nanqi, etc.), etc., and for fermentation strains, especially high-efficiency hydrogen-producing bacteria The screening research on species is relatively weak, and there is only one biohydrogen production method and its special bacteria (application number: 200910088408.8 Xing Xinhui, etc.), but the bacteria used in this method are recombinant bacteria after gene introduction, the operation is more complicated, and the fermentation conditions require Strictly anaerobic, not easy to promote

Method used

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  • Method for preparing hydrogen by fermentation through using special anaerobic clostridium pasteurianum
  • Method for preparing hydrogen by fermentation through using special anaerobic clostridium pasteurianum
  • Method for preparing hydrogen by fermentation through using special anaerobic clostridium pasteurianum

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: Determine the basal medium for strain expansion culture

[0026] The composition of the medium is: peptone 10g / L, beef extract 2.4 g / L, yeast extract 5.0g / L, Na 2 HPO 4 4g / L, L-cysteine ​​0.2 g / L, L-cysteine ​​hydrochloride monohydrate 0.5 g / L, horse serum 40-60 mL / L, distilled water 1000mL. In addition, 1 mL / L of resazurin indicator (0.2%) was added. Adjust the pH to 6.5, put every 100mL culture solution into a 250mL saline bottle (with a stopper, which can be sealed), heat the liquid in the bottle with a water bath, and blow nitrogen gas from the mouth of the bottle at the same time, until the liquid phase becomes colorless (the bottle is anaerobic). Environment), seal the bottle cap. Sterilize at 120°C for 25 minutes, dry in an oven at 60°C, inoculate the strain Clostridium pasteurianum into a bottle containing the culture medium (aseptic anaerobic operation), and culture at 37°C for 24-48 hours.

[0027] During the cultivation process, the absorba...

Embodiment 2

[0030] Embodiment 2: Selection and determination of strain fermentation substrate concentration configuration

[0031] The composition of the fermentation medium is: peptone 10g / L, beef extract 2.4 g / L, yeast extract 5.0g / L, Na 2 HPO 4 4g / L, L-cysteine ​​0.2g / L, L-cysteine ​​hydrochloride monohydrate 0.5g / L, horse serum 40-60 mL / L, resazurin (0.2%) 1mL / L, glucose 5~20g / L; adjust the initial pH=5.5~6.5; in the process of preparing the medium, use heating and boiling to expel dissolved oxygen in the liquid phase and use nitrogen to blow off the gas phase air in the bottle to ensure strict anaerobic conditions at 115 ℃ high-temperature sterilization for 25 minutes; under sterile anaerobic operation, the previously cultured Clostridium pasteurianum OD 600 The high-concentration bacterial liquid and culture medium of the inoculum with a value of 1.402 were inoculated in the culture medium in a 200ml fermentation bottle according to the inoculum amount in Table 1, and cultured s...

Embodiment 3

[0037] Embodiment 3: optimum fermentation pH is determined

[0038] The effect of initial pH value on total gas production and cell growth of Clostridium pasteuriani is shown in Tables 4 and 5. Under the condition of glucose concentration of 10g / L, the total gas production of Clostridium pasteuriani fluctuated with the change of initial pH. When the initial pH is in the range of 5.0~6.0, the total gas production and dry cell weight increase with the increase of the initial pH. When the initial pH is 6.0, the total gas production reaches a maximum of 480mL / L, and the dry cell weight reaches a maximum of 0.72g / L; when the initial pH continued to increase, both the total gas production and the dry cell weight decreased with the increase of the initial pH. This shows that under the condition of glucose mass concentration of 10g / L, the initial pH of 6.0 is the most suitable growth condition for gas production of Clostridium pasteuriani, lower or higher than this value will have a...

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Abstract

The invention provides a method for preparing hydrogen by fermentation through using special anaerobic clostridium pasteurianum. Under the sterile and anaerobic condition, anaerobic bacteria are used, a mixture consisting of peptone, beef extracts, yeast extracts, Na2HPO4, L-cysteine, L-cysteine hydrochloride monohydrate, diazoresorcinal indicators and distilled water is used as a culture medium, glucose is used as a fermentation substrate, a batch fermentation method is adopted for preparing the hydrogen, and the special anaerobic bacteria are clostridium pasteurianum JCM1408T. The culture medium consists of 10.0 to 15.0g of peptone, 2.0 to 3.0g of beef extracts, 5.0 to 10.0g of yeast extracts, 3.0 to 4.0g of Na2HPO4, 0.2 to 0.5g of L-cysteine, 0.2 to 0.5g of L-cysteine hydrochloride monohydrate, 1ml of 0.2 percent wt diazoresorcinal indicators, 40 to 60ml of horse serum and 1L of distilled water. The hydrogen can be continuously prepared.

Description

technical field [0001] The invention relates to a method for producing hydrogen by utilizing microbial anaerobic fermentation, in particular a method for producing hydrogen by utilizing highly efficient anaerobic bacteria. Background technique [0002] As a new generation of green energy, hydrogen is clean, efficient, and renewable. If it can be produced industrially, it will definitely be able to replace fossil energy such as oil and natural gas in the future and reduce the pressure on the environment. [0003] In addition, hydrogen energy is not a primary energy source and needs to be produced from hydrogen-containing compounds. At present, more than 90% of the hydrogen in the world comes from fossil combustion, and the rest is hydrogen production from water electrolysis. No matter what kind of hydrogen production method will directly or indirectly consume a large amount of fossil energy. The reserves of fossil fuels are limited and tend to be exhausted. At the same time...

Claims

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Application Information

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IPC IPC(8): C12P3/00C12R1/145
Inventor 钱春香陈璐圆王瑞兴
Owner SOUTHEAST UNIV
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