CN105441369A - Cultivation method for controlling bacillus not to generate odor - Google Patents
Cultivation method for controlling bacillus not to generate odor Download PDFInfo
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Abstract
The invention discloses a cultivation method for controlling bacillus not to generate odor, and belongs to the technical field of microbial fermentation. The method comprises the steps that glucose or glycerinum is added into a fermentation medium containing 0.5% of yeast powder, 1% of peptone and 1% of NaCl, and the concentration of the glucose or the glycerinum is controlled, so that no odor is generated in the bacillus fermentation process, wherein the pH of the fermentation medium is 7.0, Both the addition amount of the glucose and that of the glycerinum are 2-3%. The method achieves the technical effect of controlling bacillus not to generate odor, and the problem that the influence is caused by odor generated by bacillus added into the food fermentation process on the food quality is avoided.
Description
Technical field
The present invention relates to a kind of cultural method controlling genus bacillus generation stink, by adding carbon source to fermention medium and controlling the concentration of carbon source, make not produce stink in fermentation of bacillus process, belong to technical field of microbial fermentation.
Background technology
Genus bacillus is gram-positive microorganism, a kind of aerobic microbiological, and most of genus bacillus is all applied to industrial production by as important industrial strain.Be widely used in the fields such as herding, medicine, control of plant disease as subtilis, bacillus amyloliquefaciens etc., and demonstrate huge social benefit and ecological benefits.
Genus bacillus, also as the expressive host of common enzyme, is widely used in food, chemical industry, medicine and other fields.Be such as that the L-ASP of host expresses can decompose by catalysis l-asparagine with subtilis, be usually used in reducing the content of acrylamide at food processing process; For another example be applied to industrial production with the genus bacillus such as Bacillus licheniformis, subtilis, bacillus cereus and the bacillus amyloliquefaciens Pullulanase that is host expresses and amylase, substantially increase the utilization ratio of starchy material.In recent years, the genus bacillus such as bacillus amyloliquefaciens, subtilis, Bacillus licheniformis and bacillus pumilus are also applied to leavened food, as bacillus amyloliquefaciens, subtilis, Bacillus licheniformis etc. are applied in brewed spirit process the earthy that can effectively suppress in brewed spirit process.But genus bacillus can produce unpleasant, smelly taste in culturing process, if be added into the quality that can affect food in Food fermentation processes.Therefore, for this problem, the invention provides a kind of cultural method controlling genus bacillus generation stink, solve the problem that genus bacillus produces stink in liquid fermenting process.
Summary of the invention
The invention provides and a kind ofly control genus bacillus and produce the method for stink, present method, by adding glucose or glycerine and control its addition in fermention medium, makes genus bacillus not produce stink during the fermentation.
Described method uses fermention medium at thalline fermenting process; Described fermention medium, carries out proportioning by mass percent, and containing yeast powder 0.5%, peptone 1%, NaCl1%, and add glucose or glycerine wherein, regulate initial pH to be 7.0 ~ 7.2, all the other are water.
In one embodiment of the invention, the concentration of described glucose is 2% ~ 3%.
In one embodiment of the invention, the concentration of described glycerine is 2% ~ 3%.
In one embodiment of the invention, described genus bacillus be following any one or multiple: bacillus amyloliquefaciens, subtilis, bacillus thuringiensis, bacillus cereus.
In one embodiment of the invention, described thalline fermentation is undertaken by following operation: seed liquor seed liquor cultivation obtained is inoculated in fermention medium by the inoculum size of 5%, then at 37 DEG C, and the condition bottom fermentation 12 ~ 24h of 220r/min.
In one embodiment of the invention, described method first activates bacterial classification, then carries out seed liquor cultivation, re-uses fermention medium fermentation and obtain thalline.
In one embodiment of the invention, described seed liquor is cultivated operation steps and is: the single colony inoculation on picking solid medium in seed culture medium, 37 DEG C, shaking table is cultivated 12h and is obtained seed liquor under the condition of 220r/min; Described seed culture medium formula is by mass percentage as follows: yeast powder 0.5%, peptone 1%, NaCl1%, initial pH7.0.
Described method steps is as follows:
1) by the thalline streak inoculation that is kept in-80 DEG C of glycerine pipes in solid medium, be placed in 37 DEG C of constant incubators and cultivate 12h and obtain single bacterium colony.Solid medium described above formula is by mass percentage as follows: yeast powder 0.5%, peptone 1%, NaCl1%, agar 1.8% ~ 2%, pH7.0.
2) the single colony inoculation on picking solid medium is in seed culture medium, and at 37 DEG C, under the condition of 220r/min, shaking table is cultivated 12h and obtained seed liquor.Seed culture medium described above formula is by mass percentage as follows: yeast powder 0.5%, peptone 1%, NaCl1%, pH7.0.
3) by step 2) in obtain seed liquor by 5% inoculum size be inoculated in fermention medium, then at 37 DEG C, the condition bottom fermentation 12 ~ 24h of 220r/min.
The present invention also provides a kind of and controls the substratum that genus bacillus produces stink, described substratum carries out proportioning by mass percentage, containing yeast powder 0.5%, peptone 1%, NaCl1%, in addition containing glucose 2% ~ 3% or glycerine 2% ~ 3%, all the other are water.
Compared with prior art, the present invention has following advantage and beneficial effect:
The present invention by adding glucose or glycerine in fermention medium, reach and control the object that genus bacillus produces stink, its stink reduced or disappears, avoiding and genus bacillus is added in Food fermentation processes because of the impact on Food Quality that himself stink causes.
Embodiment
Below in conjunction with concrete example, the present invention is described in further detail.
Embodiment 1
1. the bacterial strain activation of bacillus thuringiensis LG1
By the bacillus thuringiensis LG1 streak inoculation that is deposited in-80 DEG C of glycerine pipes in solid medium, be placed in 37 DEG C of constant incubators and cultivate 12h and obtain single bacterium colony.The composition of solid medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, agar 20g/L, initial pH7.0.
2. the seed liquor of bacillus thuringiensis LG1 is cultivated
Single colony inoculation on picking solid medium in seed culture medium, the liquid amount 25/250mL of shake-flask seed substratum.At 37 DEG C, under the condition of 220r/min, shaking table is cultivated 12h and is obtained seed liquor.The composition of seed culture medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, initial pH7.0.
3. the thalline fermentation of bacillus thuringiensis LG1
By the seed liquor obtained in step 2 by 5% inoculum size be inoculated in fermention medium (A ~ F), then at 37 DEG C, the condition bottom fermentation of 220r/min cultivates 12 ~ 24h, evaluates thalline in fermention medium and produces stink situation.
Fermention medium composed as follows:
Fermention medium A: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, pH7.0;
Fermention medium B: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 5g/L, pH7.0;
Fermention medium C: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 5g/L, pH7.0;
Fermention medium D: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 20g/L, pH7.0.
Fermention medium E: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 20g/L, pH7.0.
Fermention medium F: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, maltose 20g/L, pH7.0.
Respectively by bacillus thuringiensis with after above-mentioned 6 kinds of fermention mediums fermentation 12h to 24h, set up 20 people's subjective appreciation groups, stink degree with 3 level evaluation fermented liquids: have obvious stink (6 ~ 10 points), there is less malodor (1 ~ 5 point), odorless (0 point).Result is as shown in table 1.
Table 1 bacillus thuringiensis LG1 fermentation ends secondary fermentation liquid Odor Evaluations cartogram
Through statistics, thalline fermenting process uses the fermented liquid of the bacillus thuringiensis LG1 of fermention medium A, B, F to have obvious stink, and through 12000r/min is centrifugal abandon supernatant after bacterium mud still have obvious stink.The fermented liquid of the bacillus thuringiensis LG1 that thalline fermenting process uses fermention medium C to obtain has less malodor, and abandons the bacterium mud after supernatant through 12000r/min high speed centrifugation and also have less malodor.And the fermented liquid odorless of the bacillus thuringiensis LG1 that thalline fermenting process uses fermention medium D, E to obtain is formed, and abandon the bacterium mud also odorless after supernatant through 12000r/min high speed centrifugation.
Embodiment 2
1. the bacterial strain activation of bacillus amyloliquefaciens JY06
By the bacillus amyloliquefaciens JY06 streak inoculation that is deposited in-80 DEG C of glycerine pipes in solid medium, be placed in 37 DEG C of constant incubators and cultivate 12h and obtain single bacterium colony.The composition of solid medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, agar 20g/L, initial pH7.0.
2. the seed liquor of bacillus amyloliquefaciens JY06 is cultivated
Single colony inoculation on picking solid medium in seed culture medium, the liquid amount 25/250mL of shake-flask seed substratum.At 37 DEG C, under the condition of 220r/min, shaking table is cultivated 12h and is obtained seed liquor.The composition of seed culture medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, initial pH7.0.
3. the thalline fermentation of bacillus amyloliquefaciens JY06
By the seed liquor obtained in step 2 by 5% inoculum size be inoculated in fermention medium (A ~ F), then at 37 DEG C, the condition bottom fermentation of 220r/min cultivates 12 ~ 24h, evaluates thalline in fermention medium and produces stink situation.
Fermention medium composed as follows:
Fermention medium A: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, pH7.2;
Fermention medium B: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 10g/L, pH7.2;
Fermention medium C: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 10g/L, pH7.2;
Fermention medium D: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 30g/L, pH7.2.
Fermention medium E: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 30g/L, pH7.2.
Fermention medium F: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, maltose 20g/L, pH7.2.
Respectively by bacillus amyloliquefaciens JY06 with after above-mentioned 6 kinds of fermention mediums fermentation 12h to 24h, set up 20 people's subjective appreciation groups, stink degree with 3 level evaluation fermented liquids: have obvious stink (6 ~ 10 points), there is less malodor (1 ~ 5 point), odorless (0 point).Result is as shown in table 2:
Table 2 bacillus amyloliquefaciens JY06 fermentation ends secondary fermentation liquid Odor Evaluations cartogram
Through statistics, thalline fermenting process uses the fermented liquid of the bacillus amyloliquefaciens JY06 of fermention medium A, B, F to have obvious stink, and through 12000r/min is centrifugal abandon supernatant after bacterium mud still have obvious stink.The fermented liquid of the bacillus amyloliquefaciens JY06 that thalline fermenting process uses fermention medium C to obtain has less malodor, and abandons the bacterium mud after supernatant through 12000r/min high speed centrifugation and also have less malodor.And the fermented liquid odorless of the bacillus amyloliquefaciens JY06 that thalline fermenting process uses fermention medium D, E to obtain is formed, and abandon the bacterium mud also odorless after supernatant through 12000r/min high speed centrifugation.
Embodiment 3
1. the bacterial strain activation of subtilis LG2
By the subtilis LG2 streak inoculation that is deposited in-80 DEG C of glycerine pipes in solid medium, be placed in 37 DEG C of constant incubators and cultivate 12h and obtain single bacterium colony.The composition of solid medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, agar 20g/L, initial pH7.0.
2. the seed liquor of subtilis LG2 is cultivated
Single colony inoculation on picking solid medium in seed culture medium, the liquid amount 25/250mL of shake-flask seed substratum.At 37 DEG C, under the condition of 220r/min, shaking table is cultivated 12h and is obtained seed liquor.The composition of seed culture medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, initial pH7.0.
3. the thalline fermentation of subtilis LG2
By the seed liquor obtained in step 2 by 5% inoculum size be inoculated in fermention medium (A ~ F), then at 37 DEG C, the condition bottom fermentation of 220r/min cultivates 12 ~ 24h.Fermention medium composed as follows:
Fermention medium A: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, pH7.0;
Fermention medium B: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 5g/L, pH7.0;
Fermention medium C: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 10g/L, pH7.0;
Fermention medium D: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 20g/L, pH7.0.
Fermention medium E: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 20g/L, pH7.2.
Fermention medium F: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, maltose 20g/L, pH7.2.
Respectively by subtilis LG2 with after above-mentioned 6 kinds of fermention mediums fermentation 12h to 24h, set up 20 people's subjective appreciation groups, stink degree with 3 level evaluation fermented liquids: have obvious stink (6 ~ 10 points), there is less malodor (1 ~ 5 point), odorless (0 point).As shown in table 3:
Table 3 subtilis LG2 fermentation ends secondary fermentation liquid Odor Evaluations cartogram
Through statistics, thalline fermenting process uses the fermented liquid of the subtilis LG2 of fermention medium A, B, F to have obvious stink, and through 12000r/min is centrifugal abandon supernatant after bacterium mud still have obvious stink.The fermented liquid of the subtilis LG2 using fermention medium C to obtain has less malodor, and abandons the bacterium mud after supernatant through 12000r/min high speed centrifugation and also have less malodor.And the fermented liquid odorless of the subtilis LG2 using fermention medium D, E to obtain is formed, and abandon the bacterium mud also odorless after supernatant through 12000r/min high speed centrifugation.
Embodiment 4
1. the bacterial strain activation of bacillus cereus LG3
By the bacillus cereus LG3 streak inoculation that is deposited in-80 DEG C of glycerine pipes in solid medium, be placed in 37 DEG C of constant incubators and cultivate 12h and obtain single bacterium colony.The composition of solid medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, agar 20g/L, initial pH7.0.
2. the seed liquor of bacillus cereus LG3 is cultivated
Single colony inoculation on picking solid medium in seed culture medium, the liquid amount 25/250mL of shake-flask seed substratum.At 37 DEG C, under the condition of 220r/min, shaking table is cultivated 12h and is obtained seed liquor.The composition of seed culture medium: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, initial pH7.0.
3. the thalline fermentation of bacillus cereus LG3
By the seed liquor obtained in step 2 by 5% inoculum size be inoculated in fermention medium (A ~ F), then at 37 DEG C, the condition bottom fermentation 12 ~ 24h of 220r/min.Fermention medium composed as follows:
Fermention medium A: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, pH7.2;
Fermention medium B: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 10g/L, pH7.2;
Fermention medium C: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 10g/L, pH7.2;
Fermention medium D: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glycerine 30g/L, pH7.2.
Fermention medium E: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, glucose 30g/L, pH7.0.
Fermention medium F: yeast powder 5g/L, peptone 10g/L, NaCl10g/L, maltose 20g/L, pH7.0.
Respectively by bacillus cereus LG3 with after above-mentioned 6 kinds of fermention mediums fermentation 12h to 24h, set up 20 people's subjective appreciation groups, stink degree with 3 level evaluation fermented liquids: have obvious stink (6 ~ 10 points), there is less malodor (1 ~ 5 point), odorless (0 point).
Table 4 bacillus cereus LG3 fermentation ends secondary fermentation liquid Odor Evaluations cartogram
Through statistics, thalline fermenting process uses the fermented liquid of the bacillus cereus LG3 of fermention medium A, B, F to have obvious stink, and through 12000r/min is centrifugal abandon supernatant after bacterium mud still have obvious stink.The fermented liquid of the bacillus cereus LG3 using fermention medium C to obtain has less malodor, and abandons the bacterium mud after supernatant through 12000r/min high speed centrifugation and also have less malodor.And the fermented liquid odorless of the bacillus cereus LG3 using fermention medium D, E to obtain is formed, and abandon the bacterium mud also odorless after supernatant through 12000r/min high speed centrifugation.
Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (10)
1. control the method that genus bacillus produces stink, it is characterized in that, described method uses fermention medium at thalline fermenting process; Described fermention medium, carries out proportioning by mass percent, and containing yeast powder 0.5%, peptone 1%, NaCl1%, and add glucose or glycerine wherein, regulate initial pH to be 7.0 ~ 7.2, all the other are water.
2. method according to claim 1, is characterized in that, the concentration of described glucose is 2% ~ 3%.
3. method according to claim 1, is characterized in that, the concentration of described glycerine is 2% ~ 3%.
4. method according to claim 1, is characterized in that, described genus bacillus be following any one or multiple: bacillus amyloliquefaciens, subtilis, bacillus thuringiensis, bacillus cereus.
5. method according to claim 1, is characterized in that, described method first activates bacterial classification, then carries out seed liquor cultivation, re-uses fermention medium and carries out thalline fermentation.
6. method according to claim 5, is characterized in that, described activation is undertaken by following operation: by the thalline streak inoculation that is kept in-80 DEG C of glycerine pipes in solid medium, is placed in 37 DEG C of constant incubators and cultivates 12h and obtain single bacterium colony.
7. method according to claim 5, is characterized in that, described actication of culture solid medium formula by mass percentage used is as follows: yeast powder 0.5%, peptone 1%, NaCl1%, agar 1.8% ~ 2%, initial pH7.0.
8. method according to claim 5, is characterized in that, described seed liquor is cultivated operation steps and is: the single colony inoculation on picking solid medium in seed culture medium, 37 DEG C, shaking table is cultivated 12h and is obtained seed liquor under the condition of 220r/min; Described seed culture medium formula is by mass percentage as follows: yeast powder 0.5%, peptone 1%, NaCl1%, initial pH7.0.
9. method according to claim 1, it is characterized in that, described thalline fermentation is undertaken by following operation: seed liquor seed liquor cultivation obtained is inoculated in fermention medium by the inoculum size of 5%, then at 37 DEG C, and the condition bottom fermentation 12 ~ 24h of 220r/min.
10. one kind controls the substratum that genus bacillus produces stink, it is characterized in that, described substratum carries out proportioning by mass percentage, containing yeast powder 0.5%, peptone 1%, NaCl1%, in addition containing glucose 2% ~ 3% or glycerine 2% ~ 3%, all the other are water.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112159786A (en) * | 2020-11-04 | 2021-01-01 | 河北科技大学 | Cr (VI) reducing strain C6, and culture condition and application thereof |
| CN112522157A (en) * | 2020-12-21 | 2021-03-19 | 南通励成生物工程有限公司 | Vitamin K for reducing fermentation of bacillus subtilis2Method of in-process malodour |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112159786A (en) * | 2020-11-04 | 2021-01-01 | 河北科技大学 | Cr (VI) reducing strain C6, and culture condition and application thereof |
| CN112159786B (en) * | 2020-11-04 | 2022-03-01 | 河北科技大学 | Cr (VI) reducing strain C6, and culture condition and application thereof |
| CN112522157A (en) * | 2020-12-21 | 2021-03-19 | 南通励成生物工程有限公司 | Vitamin K for reducing fermentation of bacillus subtilis2Method of in-process malodour |
| CN112522157B (en) * | 2020-12-21 | 2023-04-18 | 南通励成生物工程有限公司 | Vitamin K for reducing fermentation of bacillus subtilis 2 Method of in-process malodour |
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