CN106282263A - A kind of semi-continuous process produces the method for different molecular weight pulullan polysaccharide - Google Patents
A kind of semi-continuous process produces the method for different molecular weight pulullan polysaccharide Download PDFInfo
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- CN106282263A CN106282263A CN201610827587.2A CN201610827587A CN106282263A CN 106282263 A CN106282263 A CN 106282263A CN 201610827587 A CN201610827587 A CN 201610827587A CN 106282263 A CN106282263 A CN 106282263A
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- C—CHEMISTRY; METALLURGY
- 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
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
- C12P19/10—Pullulan
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Abstract
The invention discloses a kind of method that semi-continuous process produces different molecular weight pulullan polysaccharide, it is characterized in that, fermentation seed liquid is accessed in fermentation medium, employing semi continuous ferments, during the fermentation, release a certain amount of fermentation liquid, and the fresh fermentation medium of supplementary same volume, and regulate pH, continue fermentation, continue after fermentation certain time to release fermentation liquid, and the fresh fermentation medium of supplementary same volume, then regulate pH, repetitive operation, fermentation pH is different in regulation, it is thus achieved that containing the fermentation liquid of different molecular weight pulullan polysaccharide.The present invention uses semicontinuous fermentation, in conjunction with regulation different fermentations pH to reach to obtain the effect of different molecular weight pulullan polysaccharide.Compared to by batch fermentation, for obtaining the pulullan polysaccharide of different molecular weight, needed for semicontinuous fermentation, fermentation time is shorter in batches, it is thus achieved that the repetitive rate of identical pulullan polysaccharide molecular weight is higher, and uses this kind of pattern, ferments more stable.
Description
Technical field
The invention belongs to field of microbial fermentation, relate to a kind of semi-continuous process and produce the side of different molecular weight pulullan polysaccharide
Method.
Background technology
Pulullan polysaccharide has good physicochemical property, therefore possesses and is applied to food, medicine, chemical industry, environmental protection, and packaging etc. is numerous
The potentiality in field.Owing to needed for different field, pulullan polysaccharide character is different, thus cause pulullan polysaccharide molecular weight demand
Difference.Pulullan polysaccharide is caused the degree of polymerization to be changed by the difference such as strain and fermentation condition, thus have impact on its molecule
The difference of amount.It has been reported that by use different organic nitrogen sources pulullan polysaccharide weight average molecular weight can be controlled 2.8 ×
105 ~ 6.0 × 105, molecular weight dispersion index is 2.0 ~ 2.4.It addition, different by controlling the addition of K2HPO4, thus obtain
The pulullan polysaccharide of different molecular weight.
The present invention, through changing intrinsic fermentation pattern, uses different time points to release fermentation liquid, and supplementary same volume is sent out
Ferment culture medium solution, simultaneously regulation specifically fermentation pH, so that each time point is obtained to obtain pulullan polysaccharide molecular weight difference, with
Reach to obtain the purpose of different molecular weight pulullan polysaccharide.
Summary of the invention
For the problems referred to above, the present invention provides a kind of with semicontinuous fermentation, and it is general to regulate different pH production different molecular weight
Shandong orchid polysaccharide method.
The present invention solves above-mentioned technical problem and be the technical scheme is that a kind of semi-continuous process production different molecular weight is general
The method of Shandong orchid polysaccharide, it is characterised in that comprise the following steps:
(1) preparing fermentation seed liquid, be as a child followed by seed culture medium by actication of culture 6-8,28 DEG C, 180rpm cultivates 28
To 32h, obtain fermentation seed liquid；
(2) fermentation culture: seed liquor step (1) obtained accesses in fermentation medium, uses semi continuous fermentation, is sending out
During ferment, release fermentation liquid, and the fresh fermentation medium of supplementary same volume, and regulate pH, continue fermentation, sweat
Middle releasing fermentation liquid, and the fresh fermentation medium of supplementary same volume, then regulate pH, repetitive operation, fermentation pH is different in regulation,
Obtain the fermentation liquid containing different molecular weight pulullan polysaccharide；
(3) refined, the fermentation liquid containing different molecular weight pulullan polysaccharide obtained in step (2) is carried out refined extraction, obtains
Obtain the pulullan polysaccharide of different molecular weight.
Further, in described step (2), in sweat, the range of accommodation of pH is 3.5-4.5.
Further, described step (2) puts, in sweat, the 40-60% that tank volume is fermentation liquid cumulative volume.
Further, in described step (2), the inoculum concentration of seed liquor is mass percent 3 ~ 5%, uses in 5L fermentation tank and fills
There is 3L fermentation medium.
Further, the condition of described step (2) fermentation culture be ventilation be 1.25V/V, rotating speed is 400r/min, fermentation
Original ph is 5.5, starts to ferment in 24h, and temperature is 32 DEG C of cultivations, and after fermentation 24h, temperature is adjusted to 28 DEG C of fermentations.
Further, in described step (2), fermentation is carried out to 48 ~ 60h, takes out 40% fermentation liquid, then supplements same volume
Fresh fermentation medium, and regulate pH to 4.5, take out 40% fermentation liquid after continuing fermentation 48 ~ 56h, add same volume
Fresh fermentation medium, and regulate pH to 4.0, fermentation exhausts to residual sugar.
Further, the quality proportioning of seed culture medium is as follows: sucrose 100 parts, yeast 3 parts of powder of leaching, K2HPO42 parts, NaCl
2.5 parts, MgSO40.4 part, (NH4)2SO41 part, FeSO40.05 part, pH is 6.0.
Further, the quality proportioning of fermentation medium is as follows: sucrose 150 parts, peptone 5 parts, K2HPO47 parts, NaCl
2.5 parts, MgSO40.4 part, FeSO40.05 part, pH is 6.0.
It is an advantage of the current invention that: the present invention uses semicontinuous fermentation, in conjunction with regulation different fermentations pH to reach to obtain not
Effect with molecular weight pulullan polysaccharide.Compared to by batch fermentation, obtain in batches different molecular weight pulullan polysaccharide and
Saying, needed for semicontinuous fermentation, fermentation time is shorter.The repetitive rate obtaining identical pulullan polysaccharide molecular weight is higher, and employing should
The pattern of kind, ferments more stable.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearer, technical scheme will be carried out below
Clear, complete description, described embodiment is a part of embodiment of the present invention rather than whole embodiments.Based on this
Embodiment in invention, the every other reality that those of ordinary skill in the art are obtained under not making creative work premise
Execute example, broadly fall into the scope of protection of the invention.
Embodiment 1
After Aureobasidium pullulans (Aureobsidium pullulans) CGMCC NO.7055 bacterial strain activation 6 ~ 8h, by it from PDA
Slant medium is transferred in the 500mL seed culture medium that liquid amount is 100mL, and the quality proportioning of seed culture medium is as follows: sugarcane
Sugar 100 parts, yeast 3 parts of powder of leaching, K2HPO4 2 parts, NaCl 2.5 parts, MgSO4 0.4 part, (NH4) 2,SO4 1 part, FeSO4
0.05 part, pH is 6.0, and by seed liquor in above-mentioned culture medium 28 DEG C, 180rpm cultivates 32h, obtains the seed liquor of fermentation, then with
The inoculum concentration of 3% accesses in the 5L fermentation tank that liquid amount is 3L, and ventilation is 1.25V/V, and speed of agitator is 400rpm, and front 24h trains
Foster temperature is 32 DEG C, and 24h after fermentation temperature regulates to 28 DEG C.
Carry out 48h in fermentation and release 40% fermentation liquid, be denoted as " first ", supplement identical blowing volume the most wherein
Fermentation medium, and regulate pH to 4.5.Continuing fermentation 50h and release 50% fermentation liquid again, this batch fermentation liquid is designated as " second batch ".Its
Rear continuation supplements identical fermentation medium, regulates pH to 4.5, stops fermentation after fermentation 64h, produces fermentation liquid eventually and be designated as the " the 3rd
Batch ".
By extracted for above-mentioned three batch fermentation liquid refined after, draw first yield 58.2g/L, weight average molecular weight is 6.73
×105 Da.Second batch fermentation liquid yield is 67.7g/L, and weight average molecular weight is 3.94 × 105Da.3rd batch of yield is 90.3g/
L, weight average molecular weight is 2.58 × 105Da.
Embodiment 2
After Aureobasidium pullulans (Aureobsidium pullulans) CGMCC NO.7055 bacterial strain activation 6 ~ 8h, by it from PDA
Slant medium is transferred in the 500mL culture medium that liquid amount is 100mL, seed liquor 28 DEG C, and 180rpm cultivates 32h, then with 5%
Inoculum concentration access liquid amount be 3L 5L fermentation tank in, ventilation is 1.0V/V, and speed of agitator is 400rpm, front 24h cultivate
Temperature is 32 DEG C, and 24h after fermentation temperature regulates to 28 DEG C.
Carry out 56h in fermentation, release 45% fermentation liquid, be denoted as " first ", supplement identical blowing volume the most wherein
Fermentation medium, and regulate pH to 4.0.Continuing fermentation 56h and release 60% fermentation liquid again, this batch fermentation liquid is designated as " second batch ".Its
Rear continuation supplements identical fermentation medium, regulates pH to 3.5, stops fermentation after fermentation 64h, produces fermentation liquid eventually and be designated as the " the 3rd
Batch ".
Extracted refined after, draw first yield 51.7g/L, weight average molecular weight is 6.52 × 105 Da.Second is wholesale
Ferment output of fluid is 73.4g/L, and weight average molecular weight is 3.61 × 105 Da.3rd batch of yield is 98.3g/L, and weight average molecular weight is
2.45×105 Da。
Embodiment 3
After Aureobasidium pullulans (Aureobsidium pullulans) CGMCC NO.7055 bacterial strain activation 6 ~ 8h, by it from PDA
Slant medium is transferred in the 500mL culture medium that liquid amount is 100mL, seed liquor 28 DEG C, and 180rpm cultivates 28h, led
Entering in secondary seed solution 28 DEG C, 200rpm cultivates 24h.Inoculum concentration with 5% accesses in the liquid amount 30L fermentation tank as 18L, logical
Air quantity is 1.15V/V, and speed of agitator is 400rpm, and front 24h cultivation temperature is 32 DEG C, and 24h after fermentation temperature regulates to 28 DEG C.
Carry out 48h in fermentation, release 45% fermentation liquid, be denoted as " first ", supplement identical blowing volume the most wherein
Fermentation medium, and regulate pH to 3.5.Continuing fermentation 60h and release 60% fermentation liquid again, this batch fermentation liquid is designated as " second batch ".Its
Rear continuation supplements identical fermentation medium, regulates pH to 3.0, stops fermentation after fermentation 64h, produces fermentation liquid eventually and be designated as the " the 3rd
Batch ".
Extracted refined after, draw first yield 48.7g/L, weight average molecular weight is 5.72 × 105 Da.Second is wholesale
Ferment output of fluid is 77.5g/L, and weight average molecular weight is 3.78 × 105 Da.3rd batch of yield is 95.1g/L, and weight average molecular weight is
2.09×105 Da。
Various embodiments above only in order to technical scheme to be described, is not intended to limit；Although with reference to aforementioned each reality
Execute example the present invention has been described in detail, it will be understood by those within the art that: its still can to aforementioned respectively
Technical scheme described in embodiment is modified, or the most some or all of technical characteristic is carried out equivalent；And
These amendments or replacement, do not make the essence of appropriate technical solution depart from the scope of various embodiments of the present invention technical scheme.
Claims (8)
1. the method that a semi-continuous process produces different molecular weight pulullan polysaccharide, it is characterised in that comprise the following steps:
(1) preparing fermentation seed liquid, be as a child followed by seed culture medium by actication of culture 6-8,28 DEG C, 180rpm cultivates 28
To 32h, obtain fermentation seed liquid；
(2) fermentation culture: seed liquor step (1) obtained accesses in fermentation medium, uses semi continuous fermentation, is sending out
During ferment, release fermentation liquid, and the fresh fermentation medium of supplementary same volume, and regulate pH, continue fermentation, sweat
Middle releasing fermentation liquid, and the fresh fermentation medium of supplementary same volume, then regulate pH, repetitive operation, fermentation pH is different in regulation,
Obtain the fermentation liquid containing different molecular weight pulullan polysaccharide；
(3) refined, the fermentation liquid containing different molecular weight pulullan polysaccharide obtained in step (2) is carried out refined extraction, obtains
Obtain the pulullan polysaccharide of different molecular weight.
A kind of semi-continuous process the most according to claim 1 produces the method for different molecular weight pulullan polysaccharide, and its feature exists
In, in described step (2), in sweat, the range of accommodation of pH is 3.5-4.5.
A kind of semi-continuous process the most according to claim 1 produces the method for different molecular weight pulullan polysaccharide, and its feature exists
In, described step (2) is put in sweat the 40-60% that tank volume is fermentation liquid cumulative volume.
A kind of semi-continuous process the most according to claim 1 produces the method for different molecular weight pulullan polysaccharide, and its feature exists
In, in described step (2), the inoculum concentration of seed liquor is mass percent 3 ~ 5%, uses in 5L fermentation tank equipped with 3L fermentation culture
Base.
A kind of semi-continuous process the most according to claim 1 produces the method for different molecular weight pulullan polysaccharide, and its feature exists
In, the condition of described step (2) fermentation culture be ventilation be 1.25V/V, rotating speed is 400r/min, and fermentation original ph is
5.5, start to ferment in 24h, temperature is 32 DEG C of cultivations, and after fermentation 24h, temperature is adjusted to 28 DEG C of fermentations.
A kind of semi-continuous process the most according to claim 1 produces the method for different molecular weight pulullan polysaccharide, and its feature exists
In, in described step (2), fermentation is carried out to 48 ~ 60h, takes out 40% fermentation liquid, then supplements the fresh fermentation training of same volume
Support base, and regulate pH to 4.5, take out 40% fermentation liquid after continuing fermentation 48 ~ 56h, add the fresh fermentation culture of same volume
Base, and regulate pH to 4.0, fermentation exhausts to residual sugar.
7. the side of different molecular weight pulullan polysaccharide is produced according to a kind of semi-continuous process described in claim 1-6 any one
Method, it is characterised in that the quality proportioning of seed culture medium is as follows: sucrose 100 parts, yeast 3 parts of powder of leaching, K2HPO42 parts, NaCl
2.5 parts, MgSO40.4 part, (NH4)2SO41 part, FeSO40.05 part, pH is 6.0.
8. the side of different molecular weight pulullan polysaccharide is produced according to a kind of semi-continuous process described in claim 1-6 any one
Method, it is characterised in that the quality proportioning of fermentation medium is as follows: sucrose 150 parts, peptone 5 parts, K2HPO47 parts, NaCl
2.5 parts, MgSO40.4 part, FeSO40.05 part, pH is 6.0.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610827587.2A CN106282263A (en) | 2016-09-18 | 2016-09-18 | A kind of semi-continuous process produces the method for different molecular weight pulullan polysaccharide |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517347A (en) * | 2011-12-16 | 2012-06-27 | 青岛琅琊台集团股份有限公司 | Method for preparing sodium gluconate through adopting semicontinuous fermentation technology |
| CN103409480A (en) * | 2013-08-22 | 2013-11-27 | 天津北洋百川生物技术有限公司 | Method for producing Pulullans with different molecular weights |
-
2016
- 2016-09-18 CN CN201610827587.2A patent/CN106282263A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517347A (en) * | 2011-12-16 | 2012-06-27 | 青岛琅琊台集团股份有限公司 | Method for preparing sodium gluconate through adopting semicontinuous fermentation technology |
| CN103409480A (en) * | 2013-08-22 | 2013-11-27 | 天津北洋百川生物技术有限公司 | Method for producing Pulullans with different molecular weights |
Non-Patent Citations (1)
| Title |
|---|
| 孙文敬等: ""半连续发酵及其应用研究进展"", 《食品科学》 * |
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Application publication date: 20170104 |