CN110066780A - A kind of feeding alpha-galactosidase solid fermenting production technology - Google Patents
A kind of feeding alpha-galactosidase solid fermenting production technology Download PDFInfo
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- CN110066780A CN110066780A CN201910248047.2A CN201910248047A CN110066780A CN 110066780 A CN110066780 A CN 110066780A CN 201910248047 A CN201910248047 A CN 201910248047A CN 110066780 A CN110066780 A CN 110066780A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2465—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1) acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)
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- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01022—Alpha-galactosidase (3.2.1.22)
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Abstract
The invention discloses a kind of feeding alpha-galactosidase solid fermenting production technologies, including preparing seed culture medium, seed culture, prepare solid fermentation culture medium and fermented and cultured, its step are as follows, prepare seed culture medium, formula are as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO47H2O (w/v), seed culture, the seed culture medium of preparation is cultivated, prepare solid fermentation culture medium, formula are as follows: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt, MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt, Fermented and cultured carries out fermented and cultured to the culture medium prepared.The feeding alpha-galactosidase solid fermenting production technology, in tunning, based on target protein alpha-galactosidase, and the alpha-galactosidase is a kind of multi-functional glycosidase, the Flatulent factors such as raffinose or stachyose can be completely decomposed into monosaccharide, that is galactolipin, glucose and fructose have a vast market foreground in field of fodder.
Description
Technical field
The present invention relates to field of microbial culture technology, specially a kind of feeding alpha-galactosidase solid fermenting produces work
Skill.
Background technique
Alpha-galactosidase (ES 3.2.1.22) is the glycoside hydrolase of catalytically hydrolyzing alpha -1,6- galactolipin glycosidic bond.α-half
The substrate specificity of lactoside enzyme mainly has: first is that cotton seed carbohydrate oligosaccharide (Raffinose family
Oligosaccharides, RFOs), such as gossypose, stachyose and verbascose etc., they are prevalent in vegetable seeds
In, especially with content highest in seeds of leguminous plant.Second is that with α-galactose side group polysaccharide, such as galactomannans
With gala glucomannan etc..Galactomannans is primarily present in the coconut dregs of rice, guar meal and Soybean Meal, and soft wood pulp
The middle a large amount of gala glucomannan of discovery.It is generally acknowledged that the alpha-galactosidase of GH27 family has broad substrate single-minded
Property, there is preferable hydrolysing activity to cotton seed carbohydrate oligosaccharide and with α-galactose residue side chain polysaccharide substrate.α-gala
Glycosidase is helped in drift, medicine and feed industry in food, beet sugar manufacture, soft wood pulp and is all widely used.
Country's alpha-galactosidase is mainly expression bacterial strain with Pichia pastoris at present, is obtained through liquid state fermentation.Produced α-half
Raffinose or stachyose are degraded to galactolipin and sucrose by lactoside enzyme, and invertase of the sucrose again through animal intestinal secretion is hydrolyzed to
Glucose and fructose；But since weanling pig produces, sucrose enzyme level is lower, and sucrose is difficult to the utilization that is decomposed.
Summary of the invention
The purpose of the present invention is to provide a kind of feeding alpha-galactosidase solid fermenting production technologies, to solve above-mentioned back
The weanling pig currently on the market that scape technology proposes produces that sucrose enzyme level is lower, and sucrose is difficult to the problem of the utilizing that be decomposed.
To achieve the above object, the invention provides the following technical scheme: a kind of feeding alpha-galactosidase solid fermenting is raw
Production. art, including prepare seed culture medium, seed culture, prepare solid fermentation culture medium and fermented and cultured, its step are as follows:
(1) seed culture medium is prepared:
Formula are as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO4
7H2O(w/v)；
(2) seed culture:
The seed culture medium prepared in step (1) is cultivated；
(3) solid fermentation culture medium is prepared:
Formula are as follows: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermented and cultured is carried out to the culture medium prepared.
Preferably, the protein content of dregs of beans is respectively 45%, 46%, 47% and 48% in the step (1).
Preferably, the temperature of seed culture is 28 DEG C in the step (2), initial pH value 6.3, revolving speed 200r/min,
Liquid amount is 30mL, and inoculum concentration is 6.8% (v/v), fermentation time 17h.
Preferably, it is 10% (v/v) that the condition of fermented and cultured, which is inoculum concentration, in the step (4), and fermentation temperature is 28 DEG C,
PH value is 6.0, fermentation time 78h.
Compared with prior art, the beneficial effects of the present invention are: the feeding alpha-galactosidase solid fermenting production technology,
Using agricultural wastes such as corn stover, wheat stalk, straw and wheat brans as matrix, three days or so completion fermenting and producings, low energy consumption；
Downstream processes are simple, are discharged without waste and waste water, in tunning, based on target protein alpha-galactosidase, also
Cellulase, zytase, mannase and xyloglucanase enzymes isoreactivity exist, and the alpha-galactosidase is a kind of more function
Energy glycosidase, has both the activity of alpha-galactosidase and invertase, can decompose completely the Flatulent factors such as raffinose or stachyose
For monosaccharide, i.e. galactolipin, glucose and fructose, had a vast market foreground in field of fodder.
Specific embodiment
Based on the embodiments of the present invention, those of ordinary skill in the art are obtained without making creative work
The every other embodiment obtained, shall fall within the protection scope of the present invention.
The present invention provides a kind of technical solution: a kind of feeding alpha-galactosidase solid fermenting production technology, including preparation
Seed culture medium, seed culture prepare solid fermentation culture medium and fermented and cultured, and its step are as follows:
(1) seed culture medium is prepared:
Formula are as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO4
7H2O(w/v)；
(2) seed culture:
The seed culture medium prepared in step (1) is cultivated；
(3) solid fermentation culture medium is prepared:
Formula are as follows: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermented and cultured is carried out to the culture medium prepared.
The protein content of dregs of beans is respectively 45%, 46%, 47% and 48% in step (1), and setting in this way can be for difference
The dregs of beans of protein content carries out fermenting experiment.
The temperature of seed culture is 28 DEG C in step (2), initial pH value 6.3, revolving speed 200r/min, and liquid amount is
30mL, inoculum concentration are 6.8% (v/v), fermentation time 17h, and setting in this way can provide condition of culture for seed culture medium.
It is 10% (v/v) that the condition of fermented and cultured, which is inoculum concentration, in step (4), and fermentation temperature is 28 DEG C, pH value 6.0,
Fermentation time is 78h, is provided with the fermentation conducive to culture medium in this way.
Embodiment 1:
Feeding alpha-galactosidase solid fermenting production technology, its step are as follows:
(1) seed culture medium is prepared:
It is formulated as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO4
7H2O(w/v)；
Dregs of beans, protein content 45%.
(2) seed culture:
Seed culture condition: 28 DEG C of temperature, initial pH value 6.3, revolving speed 200r/min, liquid amount be 30mL (250mL's
Triangular flask), inoculum concentration is 6.8% (v/v), fermentation time 17h；
(3) solid fermentation culture medium is prepared:
Formula: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermentation condition: inoculum concentration is 10% (v/v), and 28 DEG C, pH6.0, ferment 78h；
Embodiment 2:
Feeding alpha-galactosidase solid fermenting production technology, its step are as follows:
(1) seed culture medium is prepared:
It is formulated as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO4
7H2O(w/v)；
Dregs of beans, protein content 46%.
(2) seed culture:
Seed culture condition: 28 DEG C of temperature, initial pH value 6.3, revolving speed 200r/min, liquid amount be 30mL (250mL's
Triangular flask), inoculum concentration is 6.8% (v/v), fermentation time 17h；
(3) solid fermentation culture medium is prepared:
Formula: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermentation condition: inoculum concentration is 10% (v/v), and 28 DEG C, pH6.0, ferment 78h；
Embodiment 3:
Feeding alpha-galactosidase solid fermenting production technology, its step are as follows:
(1) seed culture medium is prepared:
It is formulated as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO4
7H2O(w/v)；
Dregs of beans, protein content 47%.
(2) seed culture:
Seed culture condition: 28 DEG C of temperature, initial pH value 6.3, revolving speed 200r/min, liquid amount be 30mL (250mL's
Triangular flask), inoculum concentration is 6.8% (v/v), fermentation time 17h；
(3) solid fermentation culture medium is prepared:
Formula: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermentation condition: inoculum concentration is 10% (v/v), and 28 DEG C, pH6.0, ferment 78h；
Embodiment 4:
Feeding alpha-galactosidase solid fermenting production technology, its step are as follows:
(1) seed culture medium is prepared:
It is formulated as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO4
7H2O(w/v)；
Dregs of beans, protein content 48%.
(2) seed culture:
Seed culture condition: 28 DEG C of temperature, initial pH value 6.3, revolving speed 200r/min, liquid amount be 30mL (250mL's
Triangular flask), inoculum concentration is 6.8% (v/v), fermentation time 17h；
(3) solid fermentation culture medium is prepared:
Formula: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermentation condition: inoculum concentration is 10% (v/v), and 28 DEG C, pH6.0, ferment 78h；
As a result it tests and assesses:
As a result as shown in the table.
Table 1
The content being not described in detail in this specification belongs to the prior art well known to professional and technical personnel in the field, although
Present invention has been described in detail with reference to the aforementioned embodiments, for those skilled in the art, still can be right
Technical solution documented by foregoing embodiments is modified or equivalent replacement of some of the technical features, it is all
Within the spirit and principles in the present invention, any modification, equivalent replacement, improvement and so on should be included in protection of the invention
Within the scope of.
Claims (4)
1. a kind of feeding alpha-galactosidase solid fermenting production technology, including prepare seed culture medium, seed culture, preparation admittedly
Body fermentation medium and fermented and cultured, it is characterised in that: its step are as follows:
(1) seed culture medium is prepared:
Formula are as follows: 3.28% dregs of beans (w/v), 2% wheat bran (w/v), 0.1%KH2PO4 (w/v), 0.05%FeSO47H2O (w/
v)；
(2) seed culture:
The seed culture medium prepared in step (1) is cultivated；
(3) solid fermentation culture medium is prepared:
Formula are as follows: culture medium moisture content 60%, wheat bran 8.2137g/10g butt, bean cake powder 1.7843g/10g butt,
MnSO4H2O0.001g/10g butt, CuSO45H2O0.001g/10g butt；
(4) fermented and cultured:
Fermented and cultured is carried out to the culture medium prepared.
2. a kind of feeding alpha-galactosidase solid fermenting production technology according to claim 1, it is characterised in that: described
The protein content of dregs of beans is respectively 45%, 46%, 47% and 48% in step (1).
3. a kind of feeding alpha-galactosidase solid fermenting production technology according to claim 1, it is characterised in that: described
The temperature of seed culture is 28 DEG C in step (2), initial pH value 6.3, revolving speed 200r/min, liquid amount 30mL, inoculum concentration
For 6.8% (v/v), fermentation time 17h.
4. a kind of feeding alpha-galactosidase solid fermenting production technology according to claim 1, it is characterised in that: described
It is 10% (v/v) that the condition of fermented and cultured, which is inoculum concentration, in step (4), and fermentation temperature is 28 DEG C, pH value 6.0, fermentation time
For 78h.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110331115A (en) * | 2019-08-08 | 2019-10-15 | 河南省科学院生物研究所有限责任公司 | A kind of culture medium quickly screened for alpha-galactosidase producing strains and its application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884478A (en) * | 2006-07-10 | 2006-12-27 | 浙江省农业科学院 | Aspergillus niger variant and its fermentation process in solid medium |
| CN1900277A (en) * | 2006-07-21 | 2007-01-24 | 浙江大学 | Alpha-galactosidase solid fermenting producing method |
| WO2008102743A1 (en) * | 2007-02-19 | 2008-08-28 | Asahi Kasei Chemicals Corporation | NOVEL α-GALACTOSIDASE |
-
2019
- 2019-03-29 CN CN201910248047.2A patent/CN110066780A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884478A (en) * | 2006-07-10 | 2006-12-27 | 浙江省农业科学院 | Aspergillus niger variant and its fermentation process in solid medium |
| CN1900277A (en) * | 2006-07-21 | 2007-01-24 | 浙江大学 | Alpha-galactosidase solid fermenting producing method |
| WO2008102743A1 (en) * | 2007-02-19 | 2008-08-28 | Asahi Kasei Chemicals Corporation | NOVEL α-GALACTOSIDASE |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110331115A (en) * | 2019-08-08 | 2019-10-15 | 河南省科学院生物研究所有限责任公司 | A kind of culture medium quickly screened for alpha-galactosidase producing strains and its application |
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