CN110172407A - One plant of aspergillus oryzae for producing transfructosylase and its application - Google Patents
One plant of aspergillus oryzae for producing transfructosylase and its application Download PDFInfo
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- CN110172407A CN110172407A CN201811508345.2A CN201811508345A CN110172407A CN 110172407 A CN110172407 A CN 110172407A CN 201811508345 A CN201811508345 A CN 201811508345A CN 110172407 A CN110172407 A CN 110172407A
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- transfructosylase
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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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1051—Hexosyltransferases (2.4.1)
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/66—Aspergillus
- C12R2001/69—Aspergillus oryzae
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Abstract
The present invention relates to a kind of aspergillus oryzae mutant strain, deposit number is CCTCC NO:M2018879.The mutant strain energy high yield transfructosylase, can be widely applied to the production of the enzyme, advantageously reduces the production cost of the enzyme, accelerate it in the promotion and application of field of food industry.
Description
Technical field
The invention belongs to microbe to screen and renovation technique field, particular content is related to the rice of one plant of production transfructosylase
Aspergillus strain and its application.
Technical background
Oligofructose, also known as Fructooligosaccharides or cane fruit trisaccharide oligosaccharides mainly include ketose, Nystose, sugarcane fruit
Pentasaccharides and its mixture, it is under the action of transfructosylase, by connecting 1-3 fructosyl on the fructosyl of sucrose
And formed.Oligofructose is a kind of super bifidobacterium, Bifidobacterium can be promoted to be proliferated, and Bifidobacterium is that typically have
Beneficial bacteria is not only able to promote to digest and assimilate, treats chronic diarrhea, can also adjust endocrine and restore immune function and extensive
Multiple intestines peristalsis function, thus the symptoms such as relief of constipation.In addition, the sugariness of oligofructose is the 30-60% of sucrose, heat is very low,
Blood glucose value and insulin will not be caused to increase, on the synthesis of the hepatic glycogen controlled by insulin also without influencing, can be used as glycosuria
The health-care sweetening agent of disease and hypertensive patient.A variety of diseases can be prevented and be adjusted to oligofructose, be a kind of auxotype health care
Food does not have any toxic side effect to human body, is used in food, cosmetics, drug, feed etc. by many state approvals.
It is glucose and fructose that transfructosylase, which can be catalyzed sucrose hydrolysis, can also act on sucrose by turning fructosyl
Connect 1-3 fructosyl on fructosyl and forms oligofructose.The enzyme is widely present in higher plant and microorganism, can generate the enzyme
Plant have onion, asparagus, barley, banana, beet etc., microorganism has aspergillus niger, aspergillus oryzae, aspergillus japonicus, mould, withered grass bud
Spore bacillus etc..Transfructosylase used in industrialized production oligofructose is raw as industry mostly from microorganism at present
The important sources of transfructosylase are produced, in production practice, hair is given in the curling balling-up of mycelial growth for aspergillus niger, aspergillus oryzae
Ferment production brings inconvenience.And the transfructosylase yield of existing production bacterial strain is lower, leads to the production cost of the enzyme
It is high, limit the extensive use of the enzyme.
Summary of the invention
The present invention be solve prior art problem, provide one plant production transfructosylase aspergillus oryzae mutant strain and its
Using.Applicant screens one plant of aspergillus oryzae for itself producing transfructosylase from rotten bagasse sample
(Aspergillus oryzae) bacterial strain, it is then further screened by the method for ultraviolet mutagenesis and obtains a plant mutant bacterium, it can be big
Amplitude improves the yield of transfructosylase, is conducive to the promotion and application of the enzyme.
One aspect of the present invention provides a plant mutant strain Aspergillus oryzae FT18(Aspergillus oryzae FT18),
The China typical culture collection center of Wuhan, China Wuhan University, deposit number CCTCC are preserved on December 10th, 2018
NO:M2018879.
The present invention also provides application of the above-mentioned aspergillus oryzae mutant strain in transfructosylase production.
The present invention also provides a kind of method for producing transfructosylase, be with above-mentioned aspergillus oryzae mutant strain fermentation come
Preparation.
Applicant from rotten bagasse sample separation screening to one plant of transfructosylase superior strain aspergillus oryzae
(Aspergillus oryzae) FT01, after shake flask fermentation 5d, transfructosylase enzyme activity is up to 235 u/ in fermented supernatant fluid
ml.In order to further increase the yield of transfructosylase, applicant is bacterium germination with aspergillus oryzae FT01, is sieved by ultraviolet mutagenesis
One plant mutant strain Aspergillus oryzae of choosing acquisition (Aspergillus oryzae) FT18, after shake flask fermentation 5d, in fermented supernatant fluid
Transfructosylase enzyme activity is up to 548 u/ml, improves 133% than going out bacterium germination, unexpected technical results have been achieved.This hair
The production cost of transfructosylase will be greatly lowered in the mutant strain of bright offer, promotes transfructosylase in food service industry
In popularization and application.
Specific embodiment
The present invention has used the routine techniques and method that microbe to screen field uses.Those skilled in the art can be
On the basis of technical solution documented by the present invention, using the other conventional methods in this field, experimental program and reagent, and it is unlimited
In the restriction of the specific embodiment of the invention.
The present invention will be described in detail With reference to embodiment.
The screening and identification of the natural bacterial strain of 1 high yield transfructosylase of embodiment
Applicant's separation screening from rotten bagasse sample is inoculated with PDA plate, 30 DEG C of cultures to 29 plants of Aspergillus strains respectively
7d；The fungus block for extracting 2cm × 2cm size is inoculated in 50ml liquid submerged culture base (maltose 12%；Corn pulp 1%；Ammonium sulfate
0.5%；Potassium dihydrogen phosphate 0.35%；Dipotassium hydrogen phosphate 0.75%；Epsom salt 0.03%) in ferment, 30 DEG C of culture 5d；From
Heart thallus obtains fermented supernatant fluid.
Above-mentioned fermented supernatant fluid is subjected to transfructosylase enzyme activity determination respectively.The results show that above-mentioned 29 plants of aspergillus
Transfructosylase enzyme activity is up to 235 u/ml in the fermented supernatant fluid of bacterium.
Highest this plant of bacterium of enzyme activity is carried out ITS sequence amplification by applicant, identified, which is aspergillus oryzae
(Aspergillus oryzae), applicant is named as aspergillus oryzae FT01(Aspergillus oryzaeFT01).
1, transfructosylase enzyme activity detection method
(1) definition of enzyme-activity unit
At 40 DEG C, under the conditions of pH is 5.0, enzyme amount needed for sucrose hydrolysis per minute generates 1umol reduced sugar is defined as a work
Unit of force (U).
(2) enzyme activity determination method
(2.1) reagent and solution:
0.1mol/L disodium phosphate soln: accurately weighing disodium hydrogen phosphate dodecahydrate 35.814g, and the dissolution of 800ml water is added,
1000ml is settled to after dissolution；
0.05mol/L citric acid solution: accurately weighing monohydrate potassium 10.507g, and the dissolution of 800ml water is added, fixed after dissolution
Hold to 1000ml；
Both above-mentioned to mix, intermodulation pH is 5.0；
25% sucrose solution: it accurately weighs sucrose 25g and is dissolved in 80ml buffer, 100ml is settled to after dissolution；
10mg/ml Standard glucose solution: precision weighs DEXTROSE ANHYDROUS 1.0000g and sets stirring and dissolving in 80ml buffer, to
100ml, 4 DEG C of preservations are settled to after being completely dissolved；
DNS reagent: weighing 3,5- dinitrosalicylic acid 3.15g, adds water 500ml, stirs 5s, water-bath is to 45 DEG C.Then gradually add
Enter 100ml sodium hydroxide solution, be stirred continuously simultaneously, until solution is as clear as crystal.It is gradually added Rochelle salt again
91.0g, phenol 2.50g and anhydrous sodium sulfite 2.50g.Continue 45 DEG C of heating water baths, while water supplement 300ml, be stirred continuously,
Until the substance of addition is completely dissolved.Stop heating, after being cooled to room temperature, is settled to 1000ml with water.It is filtered with sintered glass
Device filtering, takes filtrate, and storage in a brown bottle, is kept in dark place.It is used after storing 7d at room temperature, validity period is 6 months.
(2.2) drafting of standard curve
4ml DNS reagent is added after taking 8 test tube accordings to the form below that solution is added, is sufficiently shaken up, is set and react 5min in boiling water bath, it is fast
Speed is cooled to room temperature, and is settled to 20ml with water, uses No. 0 test tube solution as control, it is molten that other each test tubes are surveyed under 540nm wavelength
The absorbance of liquid.Using absorbance as abscissa, standard curve is drawn by ordinate of glucose content.
(2.3) enzyme activity determination
Determination step is as follows:
Enzyme activity calculation formula:
Enzyme activity (U/ml or U/g)=(k × Δ A540+b)×1000×n/t/180.16
In formula: Δ A540Light absorption value difference；
The slope of k-- standard curve；
The intercept of b-- standard curve；
1000 --- conversion coefficient, mg/ml are converted to ug/ml；
T --- reaction time, 15min；
The extension rate of n-- enzyme sample；
180.16-- glucose molecule amounts, g/mol.
The mutagenesis screening of 2 transfructosylase superior strain of embodiment
Mutation randomness caused by ultraviolet mutagenesis is very strong, and it is also random for being mutated the effect of generation, it is difficult to be predicted.Therefore, in order to
Effective direct mutation is obtained, technical staff usually requires to carry out more wheel ultraviolet mutagenesis, the larger workload of screening, and there are nothings
Method obtains a possibility that effective direct mutation.But it because equipment needed for ultraviolet mutagenesis is simple, expense is few, and can obtain in a short time
Mass mutation body is obtained, therefore, it is still a kind of common mutagenic breeding method now.
The aspergillus oryzae FT01(that applicant is screened with embodiment 1Aspergillus oryzaeIt FT01) is starting strain,
Science of heredity transformation is carried out to it by ultraviolet mutagenesis method, further increases the yield of its transfructosylase.
1, lethality is determined:
Aspergillus oryzae FT01 is inoculated in PDA plate, 30 DEG C of culture 5-7d.When bacterium colony surface generate a large amount of spores when, draw 5ml without
Bacterium water elution obtains spore liquid, is resuspended after centrifugation with sterile water, is counted with blood counting chamber.A 90mm culture dish is taken, is added
(concentration is about 1 to the spore suspension that 5ml has diluted107A/mL), rotor, which is added, and stirs on magnetic stirring apparatus makes spore liquid
In uniform state.In aseptic superclean bench, irradiated with the ultraviolet lamp that power is 9w in the top of vertical range 20cm, point
Not Zhao She 30s, 45s, 60s, 75s, 90s, 105s, 120s, take irradiation after spore liquid dilute 10,100,1000 times, take 100ul
It is coated with PDA plate, is counted after 30 DEG C of culture 2-3d, is control with non-irradiated spore liquid, calculates lethality.Wherein irradiate 105s
When, lethality 95% chooses the irradiation time and carries out subsequent Mutagenesis experiments.
2, first round mutagenesis screening:
A 90mm culture dish is taken, the spore suspension that 5ml has diluted is added, and (concentration is 1 × 107Cfu/ml), be added rotor and
Stirring makes spore liquid be in uniform state on magnetic stirring apparatus.In aseptic superclean bench, with power be 9w ultraviolet lamp in
The top of vertical range 20cm is irradiated, and dilutes 1000 times after irradiating 105s, 100ul is taken to be coated with PDA plate, 30 DEG C of culture 2-3d.
The first round screens and is coated with 150 pieces of PDA plates altogether, and after 30 DEG C of culture 2-3d, each plate grows 30-50 bacterium colony,
Colonial morphology is first passed through, the mutant bacteria of short branch is screened.Applicant's picking colony form is smaller, mycelia is fine and close, periphery of bacterial colonies suede
Hair is mutant bacteria totally 96 shorter, is inoculated into PDA plate, 30 DEG C of culture 5-7d respectively.It is big that each mutant bacteria extracts 2cm × 2cm
Small fungus block is inoculated in 50ml liquid submerged culture base (maltose 12% respectively；Corn pulp 1%；Ammonium sulfate 0.5%；Biphosphate
Potassium 0.35%；Dipotassium hydrogen phosphate 0.75%；Epsom salt 0.03%) in fermentation, after 30 DEG C of culture 5d, centrifugation thallus acquisition supernatant
Liquid.The detection of transfructosylase enzyme activity is carried out to the fermented supernatant fluid liquid of acquisition respectively.Simultaneously with starting strain aspergillus oryzae
FT01 is as a control group.
The results show that in the 96 plant mutant bacterium that first round Uv-induced screening obtains, without a plant mutant bacterium fermentation supernatant
The enzyme activity of transfructosylase is higher than bacterium germination in liquid enzyme；Wherein, the enzyme activity and bacterium germination out of 81 plant mutant bacterium are substantially suitable, remaining
The enzyme activity of 15 plant mutant bacterium even generally reduces 5-9% than going out bacterium germination.
Applicant has continued 10 wheel mutagenesis screenings according to the method described above, finally obtains 1 plant of transfructosylase yield
It is significantly higher than the mutant strain of bacterium germination out, is named as aspergillus oryzae FT18(Aspergillus oryzaeFT18), which exists
It ferments in 50ml liquid submerged culture base, after 30 DEG C of culture 5d, transfructosylase in the fermented supernatant fluid that centrifugation thallus obtains
Enzyme activity is up to 548u/ml, and than go out bacterium germination raising 133%, unexpected technical results have been achieved.
Applicant is on December 10th, 2018 by aspergillus oryzae FT18(Aspergillus oryzaeFT18) in being preserved in
The China typical culture collection center of Wuhan Wuhan University, state, deposit number are CCTCC NO:M2018879.
Claims (3)
1. a kind of aspergillus oryzae mutant strain, which is characterized in that the deposit number of the aspergillus oryzae mutant strain is CCTCC NO:
M2018879。
2. application of the aspergillus oryzae mutant strain as claimed in claim 2 in production transfructosylase.
3. a kind of method for producing transfructosylase is fermented with aspergillus oryzae mutant strain described in claim 1 to prepare
's.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811508345.2A CN110172407A (en) | 2018-12-11 | 2018-12-11 | One plant of aspergillus oryzae for producing transfructosylase and its application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110904064A (en) * | 2019-11-12 | 2020-03-24 | 广西大学 | Gene sequence of fructosyl transferase and preparation method and application thereof |
| CN112941046A (en) * | 2021-02-09 | 2021-06-11 | 珠海高新区维得力生物工程有限公司 | Production process and production equipment of high-activity and high-specificity fructosyltransferase |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02182195A (en) * | 1989-01-05 | 1990-07-16 | Fuji Spinning Co Ltd | Production of inulooligosaccharide |
| CN1255947A (en) * | 1998-02-06 | 2000-06-07 | 尼斯技术公司 | Method for inverting cane sugar into glucose and fructose |
| CN102899256A (en) * | 2012-09-27 | 2013-01-30 | 量子高科(中国)生物股份有限公司 | Bacterial strain for producing fructosyl transferase and preparation method thereof |
| CN103045561A (en) * | 2012-12-17 | 2013-04-17 | 宁夏乙征生物工程有限公司 | Solid state fermentation production method for beta-D-fructofuranosidase |
| CN103937691A (en) * | 2014-04-29 | 2014-07-23 | 山东大学 | Aspergillus oryzae strain capable of producing beta-fructofuranosidase, as well as culture method and application thereof |
| CN104560936A (en) * | 2015-01-02 | 2015-04-29 | 保龄宝生物股份有限公司 | Compound immobilization method for fructosyl transferase |
| CN105112306A (en) * | 2015-09-21 | 2015-12-02 | 山东百龙创园生物科技有限公司 | Aspergillus oryzae as well as culture method and application thereof |
| CN112708567A (en) * | 2021-01-26 | 2021-04-27 | 天津科技大学 | Fructosyltransferase and high-yield strain thereof |
-
2018
- 2018-12-11 CN CN201811508345.2A patent/CN110172407A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH02182195A (en) * | 1989-01-05 | 1990-07-16 | Fuji Spinning Co Ltd | Production of inulooligosaccharide |
| CN1255947A (en) * | 1998-02-06 | 2000-06-07 | 尼斯技术公司 | Method for inverting cane sugar into glucose and fructose |
| CN102899256A (en) * | 2012-09-27 | 2013-01-30 | 量子高科(中国)生物股份有限公司 | Bacterial strain for producing fructosyl transferase and preparation method thereof |
| CN103045561A (en) * | 2012-12-17 | 2013-04-17 | 宁夏乙征生物工程有限公司 | Solid state fermentation production method for beta-D-fructofuranosidase |
| CN103937691A (en) * | 2014-04-29 | 2014-07-23 | 山东大学 | Aspergillus oryzae strain capable of producing beta-fructofuranosidase, as well as culture method and application thereof |
| CN104560936A (en) * | 2015-01-02 | 2015-04-29 | 保龄宝生物股份有限公司 | Compound immobilization method for fructosyl transferase |
| CN105112306A (en) * | 2015-09-21 | 2015-12-02 | 山东百龙创园生物科技有限公司 | Aspergillus oryzae as well as culture method and application thereof |
| CN112708567A (en) * | 2021-01-26 | 2021-04-27 | 天津科技大学 | Fructosyltransferase and high-yield strain thereof |
Non-Patent Citations (8)
| Title |
|---|
| FERNANDEZ,RC 等: "Screening of veta-fructofurampsidase-producing microorgannisms and effect of pH and temperature on enzymatic rate", 《APPLIED MICROBIOLOGY AND BIOTECHNOLOGY》 * |
| SHENGHAI WANG 等: "Enhanced production of fructosyltransferase in Aspergillus oryzae by genome shuffling", 《BIOTECHNOLOGY LETTERS》 * |
| 于国萍等: "β-呋喃果糖苷酶产生菌的筛选及产酶条件初探", 《东北农业大学学报》 * |
| 毛多斌等: "果糖基转移酶及低聚果糖生产研究进展", 《广东化工》 * |
| 覃益民等: "米曲霉GX0011 β-果糖基转移酶的分离纯化", 《高校化学工程学报》 * |
| 覃益民等: "米曲霉GX0011β-果糖基转移酶的性质研究", 《食品工业科技》 * |
| 赵秀红: "《高功能性大豆低聚糖的制备及功效应用技术》", 30 November 2018, 辽宁科学技术出版社 * |
| 马莺等: "米曲霉中提取的β-D-呋喃果糖苷酶的浓缩与纯化", 《食品科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110904064A (en) * | 2019-11-12 | 2020-03-24 | 广西大学 | Gene sequence of fructosyl transferase and preparation method and application thereof |
| CN112941046A (en) * | 2021-02-09 | 2021-06-11 | 珠海高新区维得力生物工程有限公司 | Production process and production equipment of high-activity and high-specificity fructosyltransferase |
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