CN105400821A - Method for producing hydrogen from milk product waste through fermentation - Google Patents

Method for producing hydrogen from milk product waste through fermentation Download PDF

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CN105400821A
CN105400821A CN201510920718.7A CN201510920718A CN105400821A CN 105400821 A CN105400821 A CN 105400821A CN 201510920718 A CN201510920718 A CN 201510920718A CN 105400821 A CN105400821 A CN 105400821A
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milk product
irradiation
inoculated
product waste
hydrogen
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王建龙
阴亚楠
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Tsinghua University
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P3/00—Preparation of elements or inorganic compounds except carbon dioxide

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Abstract

The invention specifically relates to a method for producing hydrogen from milk product waste through fermentation, which belongs to the technical field of hydrogen production through biological fermentation. The method comprises the following concrete steps: (1) pretreatment and pre-culture of inoculation flora; and (2) hydrogen production through microbial anaerobic fermentation with milk product waste as a substrate. The inoculation flora is mixed flora or Clostridium butyricum INET1. According to the invention, the milk product waste is used as the substrate and undergoes microbial anaerobic fermentation to produce hydrogen, so organic matters in the milk product waste is converted into clean energy hydrogen and recycling of organic pollutants in the milk product waste is realized; effects of milk product waste treatment and pollution elimination are achieved; and production of the clean energy hydrogen alleviates an energy crisis. Thus, the method provided by the invention has high economic benefits and environmental benefits.

Description

A kind of method utilizing milk product waste fermentation and hydrogen production
Technical field
The invention belongs to fermentative hydrogen production technical field, be specifically related to a kind of method utilizing milk product waste fermentation and hydrogen production.
Background technology
Hydrogen is a kind of important industrial chemicals, is all widely used in petrochemical complex, metallurgical industry, Fine Organic synthesis etc.Meanwhile, hydrogen or a kind of desirable clean energy, have the features such as combustion heat value is high, products of combustion is pollution-free.The hydrogen applied in current industry is many to be prepared by traditional method, as the non-catalytic partial oxidation of fossil oil, and the steam restructuring of methane, the electrolysis etc. of water.These methods not only consume energy high, and cost is high, also have certain pollution to environment.And utilize microbial fermentation processes to prepare hydrogen, then can promote the environmental benefit of hydrogen, realize Sustainable Production.
Fermentative hydrogen production can gas chromatography be substrate, and that relatively commonly uses in research at present has carbohydrate, organic waste water, organic solid castoff etc.With simple substance carbohydrate for substrate can obtain higher hydrogen-producing speed and substrate utilization ratio, but cost is higher; Take organic waste as substrate, not only can reduce the cost of fermenting process of preparing hydrogen, the effect that organic waste are administered can also be played.
Along with the growth of national economy and the raising of living standards of the people, the demand of people to milk-product also increases thereupon.The main component of milk-product factory effluent comprises lactose, protein, lipid etc., has the advantages that organic concentration content is high, biodegradability is strong.In recent years, along with the growth of milk-product sales volume and the exploitation of product diversity, the process of milk product waste faces the problems such as wastewater flow rate is large, variation water quality is large.
Utilize milk product waste organic content high, the feature that biodegradability is strong, it can be used as fermentable to produce the substrate of hydrogen, not only can reduce the cost producing hydrogen, also contribute to realizing milk product waste resource utilization, reach the object turned waste into wealth.
Summary of the invention
The object of this invention is to provide a kind of method utilizing milk product waste fermentation and hydrogen production, is below the detailed description of technical solution of the present invention:
Utilize a method for milk product waste fermentation and hydrogen production, comprise the following steps:
(1) pre-treatment and the preculture of flora is inoculated;
(2) take milk product waste as substrate, carry out microbiological anaerobic fermentation and hydrogen production.
Described inoculation flora is mixed bacterial or clostridium butylicum INET1.
(1) when inoculation flora is mixed bacterial, pretreatment process is: with the gamma-ray irradiation process anaerobically digested sludge of 5kGy dosage, radioactive source is 60co source, obtains the digested sludge after irradiation;
Preculture method is: by the digested sludge after irradiation with 1:10 inoculation than being inoculated in pre-culture medium, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtain the mixed bacterial strong to irradiation tolerance;
Described pre-culture medium component is: lactose 20g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
(2) when inoculation flora is clostridium butylicum INET1, pretreatment process is: with the gamma-ray irradiation process anaerobically digested sludge of 5kGy dosage, radioactive source is 60co source, obtains the digested sludge after irradiation;
Preculture method is: by the digested sludge after irradiation with 1:10 inoculation than being inoculated in pre-culture medium, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtain the mixed bacterial strong to irradiation tolerance; Mixed bacterial is inoculated in isolation medium in the mode of coating and line, is separated through 3-5 time, obtains the single bacterium colony of circle, oyster white, neat in edge, be clostridium butylicum INET1;
Described pre-culture medium component is: glucose 50g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; Isolation medium component is: glucose 50g/L, peptone 10g/L, yeast powder 0.5g/L, agar powder 15-20g/L, nutritive medium 10mL/100mL; The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
The concrete operations of described hydrogen production through anaerobic fermentation are, the inoculation flora strong to irradiation tolerance enrichment obtained is inoculated than inoculating into reactor with 1:10; With nitrogen stripping reactor before reaction starts, for reaction provides anaerobic condition; Leavening temperature is 36 DEG C, 100r/min constant-temperature shaking culture, until stop aerogenesis.
In the organic components of milk product waste of the present invention, lactose mass percentage is 15% ~ 30%.
Beneficial effect of the present invention is: using milk product waste as substrate, by microbiological anaerobic fermentation and hydrogen production, the organism in waste water can be transformed into clean energy hydrogen on the one hand, realize the resource utilization of Organic Pollutants in Wastewater; Also reach the effect of administering waste water, decontamination on the other hand; The production of clean energy hydrogen also has the effect of alleviating energy crisis.Therefore, this invention has higher economic benefit and environmental benefit.
Accompanying drawing explanation
Fig. 1 is in embodiment 1, and mixed bacterial utilizes hydrogen output-time changing curve figure during lactose product hydrogen.
Fig. 2 is in embodiment 2, and embedding mixed bacterial utilizes hydrogen output-time changing curve figure during lactose product hydrogen.
Fig. 3 is in embodiment 3, and clostridium butylicum INET1 utilizes hydrogen output-time changing curve figure during lactose product hydrogen.
Fig. 4 is in embodiment 4, and mixed bacterial utilizes hydrogen output-time changing curve figure during milk product waste product hydrogen.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described.
The anaerobically digested sludge used in the embodiment of the present invention takes from the mud primary digestion pond of Beijing's municipal sewage plant.
Embodiment 1
Be substrate with lactose, utilize mixed bacterial to carry out fermentation and hydrogen production, concrete steps are as follows:
Digested sludge is placed in 1L brown port grinding bottle, at ambient temperature warp 60co is the gamma-ray irradiation of radioactive source, and irradiation dose is 5kGy.Digested sludge after irradiation is inoculated in pre-culture medium with the inoculation of 1:10 ratio, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtains the mixed bacterial strong to irradiation tolerance.Wherein, pre-culture medium component is: lactose 20g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
Getting mixed bacterial bacterium liquid that 10mL preculture obtains, to be inoculated in volume be in 100mL, COD=5g/L lactose fermentation substratum, and the initial pH of regulator solution is 7.0.Nitrogen stripping reactor before reaction, inspection resistance to air loss is also driven away in bottle after oxygen, and reaction flask is placed in water bath with thermostatic control shaking table, with 36 DEG C, 100r/min conditioned response, interval 2h records gas production rate.
After 36h, reaction terminates, and obtaining largest cumulative hydrogen output is 126mL/100mL, and gas producing efficiency is 275mL/gCOD.
Embodiment 2
Be substrate with lactose, the mixed bacterial utilizing PVA-Lalgine to embed carries out fermentation and hydrogen production, and concrete steps are as follows:
Digested sludge is placed in 1L brown port grinding bottle, at ambient temperature warp 60co is the gamma-ray irradiation of radioactive source, and irradiation dose is 5kGy.Digested sludge after irradiation is inoculated in pre-culture medium with the inoculation of 1:10 ratio, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtains the mixed bacterial strong to irradiation tolerance.Wherein, pre-culture medium component is: lactose 20g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
Mixed bacterial bacterium liquid 10mL preculture obtained obtains mud flco after 5000r/min is centrifugal, and after adopting PVA-Lalgine entrapping method to embed, being inoculated in volume is in 100mL, COD=5g/L lactose fermentation substratum, and the initial pH of regulator solution is 7.0.Nitrogen stripping reactor before reaction, inspection resistance to air loss is also driven away in bottle after oxygen, and reaction flask is placed in water bath with thermostatic control shaking table, with 36 DEG C, 100r/min conditioned response, interval 2h records gas production rate.
After 24h, reaction terminates, and obtaining largest cumulative hydrogen output is 130mL/100mL, and gas producing efficiency is 309mL/gCOD.
Embodiment 3
Be substrate with lactose, utilize clostridium butylicum INET1 to carry out fermentation and hydrogen production, concrete steps are as follows:
Digested sludge is placed in 1L brown port grinding bottle, at ambient temperature warp 60co is the gamma-ray irradiation of radioactive source, and irradiation dose is 5kGy.Digested sludge after irradiation is inoculated in pre-culture medium with the inoculation of 1:10 ratio, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtains the mixed bacterial strong to irradiation tolerance; Mixed bacterial is inoculated in isolation medium in the mode of coating and line, is separated through 3-5 time, obtains the single bacterium colony of circle, oyster white, neat in edge, be clostridium butylicum INET1; Wherein, pre-culture medium component is: glucose 50g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; Isolation medium component is: glucose 50g/L, peptone 10g/L, yeast powder 0.5g/L, agar powder 15-20g/L, nutritive medium 10mL/100mL; The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
Bacterial strain grows into logarithmic phase through 8-10h, now, getting bacterium liquid that 10mL is in logarithmic phase, to be inoculated in volume be 100mL, in COD=10g/L lactose fermentation substratum, the initial pH of regulator solution is 7.0, and nitrogen stripping inspection resistance to air loss also to be driven away in reaction flask after oxygen, reaction flask is placed in water bath with thermostatic control shaking table, with 36 DEG C, 100r/min conditioned response, interval 2h records gas production rate.
After 40h, reaction terminates, and obtaining largest cumulative hydrogen output is 180mL/100mL, and gas producing efficiency is 213mL/gCOD.
Embodiment 4
Be substrate with milk product waste, utilize mixed bacterial to carry out fermentation and hydrogen production, concrete steps are as follows:
Digested sludge is placed in 1L brown port grinding bottle, at ambient temperature warp 60co is the gamma-ray irradiation of radioactive source, and irradiation dose is 5kGy.Digested sludge after irradiation is inoculated in pre-culture medium with the inoculation of 1:10 ratio, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtains the mixed bacterial strong to irradiation tolerance.Wherein, pre-culture medium component is: lactose 20g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
Get mixed bacterial bacterium liquid that 10mL preculture obtains to be inoculated in 100mL milk product waste (COD concentration is 2890mg/L, lactose concn is 0.75g/L, protein concn 1.56g/L) in, nitrogen stripping reactor before reaction, inspection resistance to air loss is also driven away in bottle after oxygen, reaction flask is placed in water bath with thermostatic control shaking table, with 36 DEG C, 100r/min conditioned response, interval 2h records gas production rate.
After 40h, reaction terminates, and obtaining largest cumulative hydrogen output is 62mL/100mL.

Claims (6)

1. utilize a method for milk product waste fermentation and hydrogen production, it is characterized in that, comprise the following steps:
(1) pre-treatment and the preculture of flora is inoculated;
(2) take milk product waste as substrate, carry out microbiological anaerobic fermentation and hydrogen production.
2. method according to claim 1, is characterized in that, described inoculation flora is mixed bacterial or clostridium butylicum INET1.
3. method according to claim 1, is characterized in that,
When inoculation flora is mixed bacterial, pretreatment process is: with the gamma-ray irradiation process anaerobically digested sludge of 5kGy dosage, radioactive source is 60co source, obtains the digested sludge after irradiation;
Preculture method is: the digested sludge after irradiation inoculated than being inoculated in pre-culture medium with 1:10, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtains the mixed bacterial strong to irradiation tolerance;
When inoculation flora is clostridium butylicum INET1, pretreatment process is: with the gamma-ray irradiation process anaerobically digested sludge of 5kGy dosage, radioactive source is 60co source, obtains the digested sludge after irradiation;
Preculture method is: the digested sludge after irradiation inoculated than being inoculated in pre-culture medium with 1:10, at 36 DEG C, constant-temperature shaking culture 20-48h under anaerobic condition, obtains the mixed bacterial strong to irradiation tolerance; Mixed bacterial is inoculated in isolation medium in the mode of coating and line, is separated through 3-5 time, obtains the single bacterium colony of circle, oyster white, neat in edge, be clostridium butylicum INET1.
4. method according to claim 1, is characterized in that,
When inoculation flora is mixed bacterial, pre-culture medium component is: lactose 20g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL;
When inoculation flora is clostridium butylicum INET1, pre-culture medium component is: glucose 50g/L, peptone 10g/L, yeast powder 0.5g/L, nutritive medium 10mL/100mL; Isolation medium component is: glucose 50g/L, peptone 10g/L, yeast powder 0.5g/L, agar powder 15-20g/L, nutritive medium 10mL/100mL;
The component of described nutritive medium is: NaHCO 340g/L, NH 4cl5g/L, NaH 2pO 42H 2o5g/L, K 2hPO 43H 2o5g/L, FeSO 47H 2o0.25g/L, MgCl 26H 2o0.085g/L, NiCl 26H 2o0.004g/L.
5. method according to claim 1, is characterized in that, the concrete operations of described hydrogen production through anaerobic fermentation are, the inoculation flora strong to irradiation tolerance enrichment obtained is inoculated than inoculating into reactor with 1:10; With nitrogen stripping reactor before reaction starts, for reaction provides anaerobic condition; Leavening temperature is 36 DEG C, 100r/min constant-temperature shaking culture, until stop aerogenesis.
6. method according to claim 1, is characterized in that, in the organic components of described milk product waste, lactose mass percentage is 15%-30%.
CN201510920718.7A 2015-12-11 2015-12-11 Method for producing hydrogen from milk product waste through fermentation Pending CN105400821A (en)

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Citations (3)

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Publication number Priority date Publication date Assignee Title
CN102747106A (en) * 2012-07-02 2012-10-24 同济大学 Method for increasing protein bio-hydrogen production
CN103508611A (en) * 2012-06-15 2014-01-15 苏州腾辉环保科技有限公司 Process for treating dairy waste water
CN103667352A (en) * 2012-08-31 2014-03-26 青岛嘉能节能环保技术有限公司 Method for performing biological hydrogen production by utilizing organic wastewater

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103508611A (en) * 2012-06-15 2014-01-15 苏州腾辉环保科技有限公司 Process for treating dairy waste water
CN102747106A (en) * 2012-07-02 2012-10-24 同济大学 Method for increasing protein bio-hydrogen production
CN103667352A (en) * 2012-08-31 2014-03-26 青岛嘉能节能环保技术有限公司 Method for performing biological hydrogen production by utilizing organic wastewater

Non-Patent Citations (2)

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Title
王博 等: "γ辐照预处理促进剩余污泥发酵产氢的研究", 《中国环境科学》 *
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Application publication date: 20160316