CN102703532A - Method for preparing acetic acid through two-step fermentation of whey - Google Patents
Method for preparing acetic acid through two-step fermentation of whey Download PDFInfo
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- CN102703532A CN102703532A CN2012102122911A CN201210212291A CN102703532A CN 102703532 A CN102703532 A CN 102703532A CN 2012102122911 A CN2012102122911 A CN 2012102122911A CN 201210212291 A CN201210212291 A CN 201210212291A CN 102703532 A CN102703532 A CN 102703532A
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Abstract
The invention provides a method for preparing acetic acid through two-step fermentation of whey. The whey is used as the original material, lactobacillus plantarum and propionibacterium are used as starting strains, and two-step fermentation temperature, inoculation amount, rotating speed, liquid filling amount and fermentation time are studied.
Description
Technical field
The invention belongs to fermentation technical field, be specifically related to a kind of method of utilizing the whey two-step fermentation to prepare acetate.
Background technology
Acetate is a kind of important Organic Chemicals and the solvent in the chemical reaction, can be widely used in industry such as agricultural chemicals, medicine, synthetic materials, chemical fibre and tobacco, in national economy, occupies considerable effect.The working method of acetate has many kinds, at present domestic commonly used be acetaldehyde oxidation and methanol carbonyl synthesis method.
Whey is the sub product of industrial production cheese and casein food grade, and every production 1 kg cheese and 1 kg casein food grade can produce 9 kg and 27 kg wheys respectively, and therefore, though cheese of China and casein food grade production still are in developmental stage, still can produce a large amount of wheys every year.Lactose-content is high in the whey, and is nutritious, and the waste to resource is not only in a large amount of dischargings of whey, especially environment has been caused great destruction.Therefore, the rational exploitation and utilization whey has not only been practiced thrift resource, also environment has been played provide protection.
This patent is raw material with the whey, is starting strain with plant lactobacillus and propionibacterium, and the leavening temperature, inoculum size, rotating speed, liquid amount and the fermentation time that influence two-step fermentation production acetate are studied.
Summary of the invention
The invention provides a kind of method of utilizing the whey two-step fermentation to prepare acetate; This patent is raw material with the whey; With plant lactobacillus and propionibacterium is starting strain, and the leavening temperature, inoculum size, rotating speed, liquid amount and the fermentation time that influence two-step fermentation production acetate are studied.
1. method of utilizing the whey two-step fermentation to prepare acetate; It is characterized in that fermentation carries out in two steps: in 250 mL triangular flasks, pack into the fermention medium of certain volume of the first step; Insert a certain amount of plant lactobacillus seed liquor in fermention medium; Cultivate with certain rotating speed and temperature bottom fermentation regular hour at different original ph condition bottom fermentations respectively, the lactose fermentation in the whey is become lactic acid; Second step was inserted propionibacterium in the fermented liquid that the first step obtains with certain inoculum size, continued at certain temperature, rotating speed bottom fermentation certain hour lactobacillus ferment to be become acetate.
2. method according to claim 1 is characterized in that, fermention medium (g/L): whey 40, peptone 15, yeast extract paste 5, NH
4HCO
33, MgSO
43, K
2HPO
43, pH5.0.
3. method according to claim 1 is characterized in that the first step best liquid amount that ferments is 150 mL/250 mL triangular flasks.
4. method according to claim 1 is characterized in that the first step fermentation optimum inoculation amount is 5%.
5. method according to claim 1 is characterized in that the first step fermentation optimum fermentation temp is 37 ℃.
6. method according to claim 1 is characterized in that the first step fermentation optimum revolution is 50 rpm/min.
7. method according to claim 1 is characterized in that the first step best fermentation time that ferments is 42 h.
8. method according to claim 1 is characterized in that second step fermentation optimum inoculation amount is 4%.
9. method according to claim 1 is characterized in that second step fermentation optimum fermentation temp is 37 ℃.
10. method according to claim 1 is characterized in that second step fermentation optimum revolution is 60 rpm/min.
11. method according to claim 1 is characterized in that the best fermentation time of second step fermentation is 48 h.
Description of drawings
The different liquid amounts of Fig. 1 are to the influence of the first step lactic acid-fermenting output.
Fig. 2 different vaccination amount is to the influence of the first step lactic acid-fermenting output.
Fig. 3 different fermentations temperature is to the influence of the first step lactic acid-fermenting output.
Fig. 4 different rotating speeds is to the influence of the first step lactic acid-fermenting output.
Fig. 5 different fermentations time is to the influence of the first step lactic acid-fermenting output.
Fig. 6 different vaccination amount is to the influence of second step fermentation yield of acetic acid.
Fig. 7 different fermentations temperature is to the influence of second step fermentation yield of acetic acid.
Fig. 8 different rotating speeds is to the influence of second step fermentation yield of acetic acid.
The influence of Fig. 9 different fermentations time second step fermentation yield of acetic acid.
Embodiment
Following embodiment elaborates to the present invention, but to not restriction of the present invention.
This case study on implementation is explained the influence of different liquid amounts to the first step lactic acid-fermenting output, fermention medium (g/L): whey 40, peptone 15, yeast extract paste 5, NH
4HCO
33, MgSO
43, K
2HPO
43, pH5.0.The substratum that configures 60 mL, 90mL, 120mL, 150 mL, 180 mL are packed in the 250 mL triangular flasks into 121 ℃ of sterilization 20 min.In each triangular flask, insert 5% plant lactobacillus seed, in 37 ℃, 60 rpm/min bottom fermentations, 48 h, after fermentation stops, measure the content (with reference to GB/T 12456-2008) of lactic acid, it is as shown in Figure 1 to the influence of lactic acid production to obtain liquid amount.
Plant lactobacillus is a kind of anerobes, therefore increases liquid amount, helps fermentation.Can be known that by Fig. 1 along with the increase of liquid amount, lactic acid production significantly increased before this, but when liquid amount was 180 mL/250 mL triangular flasks, output descended on the contrary, therefore selecting 150 mL/250 mL triangular flasks is best liquid amount.
This case study on implementation explanation different vaccination amount is to the influence of the first step lactic acid production, fermention medium (g/L): whey 40, peptone 15, yeast extract paste 5, NH
4HCO
33, MgSO
43, K
2HPO
43, pH5.0.The substratum that configures 150 mL are packed in the 250 mL triangular flasks into 121 ℃ of sterilization 20 min.In each triangular flask, insert 2%, 3%, 4%, 5%, 6%, 7% plant lactobacillus seed respectively; In 37 ℃, 60 rpm/min bottom fermentations, 48 h; After fermentation stops, measure the content (with reference to GB/T 12456-2008) of lactic acid, it is as shown in Figure 2 to the influence of lactic acid production to obtain inoculum size.
Can know that by Fig. 2 when inoculum size was 2-5%, lactic acid production increased fast with the increase of inoculum size, when inoculum size greater than 5% the time, along with the increase of inoculum size, lactic acid production also increases to some extent, but changes not quite, therefore, selects 5% to be optimum inoculation amount.
This case study on implementation explanation different fermentations temperature is to the influence of the first step lactic acid production, fermention medium (g/L): whey 40, peptone 15, yeast extract paste 5, NH
4HCO
33, MgSO
43, K
2HPO
43, pH5.0.The substratum that configures 150 mL are packed in the 250 mL triangular flasks into 121 ℃ of sterilization 20 min.In each triangular flask, insert 5% plant lactobacillus seed; Respectively at 28 ℃, 30 ℃, 32 ℃, 35 ℃, 37 ℃, 40 ℃; 60 rpm/min bottom fermentations, 48 h; After fermentation stops, measure the content (with reference to GB/T 12456-2008) of lactic acid, it is as shown in Figure 3 to the influence of lactic acid production to obtain the different fermentations temperature.
Can be known that by Fig. 3 plant lactobacillus output when low temperature is lower, along with the rising of temperature, lactic acid production increases gradually, and output reaches maximum in the time of 37 ℃, is 2.58 g, and output descends to some extent afterwards, therefore selects 37 ℃ to be optimum fermentation temp.
Embodiment 4
This case study on implementation explanation different rotating speeds is to the influence of the first step lactic acid production, fermention medium (g/L): whey 40, peptone 15, yeast extract paste 5, NH
4HCO
33, MgSO
43, K
2HPO
43, pH5.0.The substratum that configures 150 mL are packed in the 250 mL triangular flasks into 121 ℃ of sterilization 20 min.In each triangular flask, insert 5% plant lactobacillus seed; Respectively at 40,60,80,100,120 rpm/min, 37 ℃ of bottom fermentation 48 h are after fermentation stops; Measure the content (with reference to GB/T 12456-2008) of lactic acid, it is as shown in Figure 4 to the influence of lactic acid production to obtain rotating speed.
Plant lactobacillus is an anerobes, and the rotating speed I helps fermentation and carries out to reduce dissolved oxygen.Can be known that by Fig. 4 to 60 rpm/min, lactic acid production is higher from 40 rpm/min, and in rising trend, to 120 rpm/min, lactic acid production reduces thereupon the raising of rotating speed gradually from 60 rpm/min.Therefore, select 60 rpm/min as optimum revolution.
Embodiment 5
This case study on implementation explanation different fermentations time is to the influence of the first step lactic acid production, fermention medium (g/L): whey 40, peptone 15, yeast extract paste 5, NH
4HCO
33, MgSO
43, K
2HPO
43, pH5.0.The substratum that configures 150 mL are packed in the 250 mL triangular flasks into 121 ℃ of sterilization 20 min.In each triangular flask, insert 5% plant lactobacillus seed; Respectively at 60 rpm/min, 37 ℃ of ferment 24 h, 30 h, 36 h, 42 h, 48 h; After fermentation stops, measure the content (with reference to GB/T 12456-2008) of lactic acid, it is as shown in Figure 5 to the influence of lactic acid production to obtain rotating speed.
Can know that by Fig. 5 along with the prolongation of fermentation time, the output of lactic acid increases gradually, after 42 h,, change not quite, therefore, select 42 h as the righttest fermentation time though increase to some extent.
This case study on implementation explanation different vaccination amount is to the influence of second step fermentation yield of acetic acid; In the fermented liquid of same batch of the first step fermentation, insert 2%, 3%, 4%, 5%, 6%, 7% propionibacterium seed respectively; In 60 rpm/min, 37 ℃ of fermentation 48 h; After fermentation stops, measure the content (HPLC method) of acetate, it is as shown in Figure 6 to the influence of yield of acetic acid to obtain the different vaccination amount.
Can know that by Fig. 6 when inoculum size from 2% to 4% changed, along with the increase of inoculum size, yield of acetic acid increased sharply, and from 4% to 7%, output is also in continuous increase, but variation is little, therefore selects 4% as optimum inoculation amount.
Embodiment 7
This case study on implementation explanation different fermentations temperature is to the influence of second step fermentation yield of acetic acid; In the fermented liquid of same batch of the first step fermentation, insert 4% propionibacterium seed respectively; Respectively at 30 ℃, 32 ℃, 35 ℃, 37 ℃, 40 ℃, 60 rpm/min, fermentation 48 h are after fermentation stops; Measure the content (HPLC method) of acetate, it is as shown in Figure 7 to the influence of yield of acetic acid to obtain the different fermentations temperature.
Can know that by Fig. 7 along with the rising of leavening temperature, the output of acetate increases gradually, but output begins to descend to some extent after 37 ℃, so select 37 ℃ of the righttest leavening temperatures of conduct.
Embodiment 8
This case study on implementation explanation different rotating speeds is to the influence of second step fermentation yield of acetic acid; In the fermented liquid of same batch of the first step fermentation, insert 4% propionibacterium seed respectively; Respectively at 40,50,60,70,80 rpm/min, 37 ℃ of fermentation 48 h are after fermentation stops; Measure the content (HPLC method) of acetate, it is as shown in Figure 8 to the influence of yield of acetic acid to obtain different rotating speeds.
Can know that by Fig. 8 to 50 rpm/min, yield of acetic acid increases with the rising of rotating speed from 40 rpm/min; To 80 rpm/min, yield of acetic acid reduces with the rising of rotating speed from 50 rpm/min, therefore, selects 50 rpm/min as optimum revolution.
Embodiment 9
This case study on implementation explanation different fermentations time is to the influence of second step fermentation yield of acetic acid; In the fermented liquid of same batch of the first step fermentation, insert 4% propionibacterium seed respectively; 24,30,36,42,48 h ferment respectively under 50rpm/min, 37 ℃; After fermentation stops, measure the content (HPLC method) of acetate, it is as shown in Figure 9 to the influence of yield of acetic acid to obtain the different fermentations time.
Can know that by Fig. 9 along with the prolongation of fermentation time, the output of acetate constantly increases, but speedup slows down behind 48 h, therefore, select 48 h as the righttest fermentation time.
Claims (10)
1. method of utilizing the whey two-step fermentation to prepare acetate; It is characterized in that fermentation carries out in two steps: in 250 mL triangular flasks, pack into the fermention medium of certain volume of the first step; Insert a certain amount of plant lactobacillus seed liquor in fermention medium; Cultivate with certain rotating speed and temperature bottom fermentation regular hour at different original ph condition bottom fermentations respectively, the lactose fermentation in the whey is become lactic acid; Second step was inserted propionibacterium in the fermented liquid that the first step obtains with certain inoculum size, continued at certain temperature, rotating speed bottom fermentation certain hour lactobacillus ferment to be become acetate.
2. method according to claim 1 is characterized in that fermention medium: whey 40 g/L, peptone 15 g/L, yeast extract paste 5 g/L, NH
4HCO
33 g/L, MgSO
43 g/L, K
2HPO
43 g/L, pH5.0.
3. method according to claim 1 is characterized in that the first step fermentation inoculum size is 5%.
4. method according to claim 1 is characterized in that the first step fermentation leavening temperature is 37 ℃.
5. method according to claim 1 is characterized in that the first step fermentation rotating speed is 50 rpm/min.
6. method according to claim 1 is characterized in that the first step fermentation fermentation time is 42 h.
7. method according to claim 1 is characterized in that second step fermentation inoculum size is 4%.
8. method according to claim 1 is characterized in that second step fermentation leavening temperature is 37 ℃.
9. method according to claim 1 is characterized in that second step fermentation rotating speed is 60 rpm/min.
10. method according to claim 1 is characterized in that second step fermentation fermentation time is 48 h.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101748083A (en) * | 2008-12-11 | 2010-06-23 | 吉林省农业科学院 | Lactobacillus plantarum ferment and the preparation method and special strain thereof |
| CN101748082A (en) * | 2008-12-11 | 2010-06-23 | 吉林省农业科学院 | Lactobacillus leavening agent, preparation method thereof and special bacterial strain |
-
2012
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101748083A (en) * | 2008-12-11 | 2010-06-23 | 吉林省农业科学院 | Lactobacillus plantarum ferment and the preparation method and special strain thereof |
| CN101748082A (en) * | 2008-12-11 | 2010-06-23 | 吉林省农业科学院 | Lactobacillus leavening agent, preparation method thereof and special bacterial strain |
Non-Patent Citations (2)
| Title |
|---|
| 吕海棠等: "乳清发酵法制备乙酸", 《安徽农业科学》 * |
| 韩爱霞等: "乳清两步发酵法制取环保型融雪剂醋酸钙镁盐", 《无机盐工业》 * |
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Application publication date: 20121003 |