CN110106090A - A kind of Neuraspora crassa strain and its application - Google Patents
A kind of Neuraspora crassa strain and its application Download PDFInfo
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Abstract
The invention belongs to technical field of food biotechnology, a kind of Neuraspora crassa strain and its application are specifically disclosed.The bacterial strain is Neuraspora crassa Neurospora crassa, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and the deposit date is on 01 16th, 2019, deposit number was CGMCC NO:3.19233.Neuraspora crassa strain LY03 fermentation with carriers such as corn flour or starch is prepared by mixing into fermentation bacteria cake after spreading cultivation, and will make red bacterium bean curd in fermentation bacteria cake access bean dregs.The ingredients such as corn flour, the starch added in fermentation bacteria cake are conducive to the dispersion of fermentation strain mycelia, reinforcing fiber enzyme activity, reduce the carbohydrate contents such as fiber, the polysaccharide in bean dregs, increase the content of total protein in finished product, improve mouthfeel.The present invention overcomes traditional handicraft fermentation strains to present by nature, and bacterial strain batch is different, the small disadvantage of unstable quality, production scale, while having widened soybean and its attached material production industrial chain, has a good application prospect.
Description
Technical field
The invention belongs to technical field of food biotechnology, and in particular to a kind of Neuraspora crassa strain and its application.
Background technique
" red bacterium bean curd " originates in Fujian Province Wuping County, history have how long, have no way of investigating, only know before monosodium glutamate appearance, warp
Often be taken as monosodium glutamate come using.Rapidly, 3~5d's fermentation of red bacterium bean curd can ferment completion, and " red bacterium bean curd " after fermentation has
Sweet fresh, sweet mouthfeel, therefore locals uses as monosodium glutamate, is food fresh adding flavouring.In addition, it was found that " red bacterium beans
It is rotten " also there is important health-care effect, it can improve human body intestinal canal function, increase the moving property of enteron aisle, and can adsorb intestines
Grease in road simultaneously excretes, and clears up the toxin in enteron aisle, mitigates the removing toxic substances burden of liver, is one to three high crowds
A very good food.In addition, the raw material that " red bacterium bean curd " uses is discarded bean dregs, waste utilization is realized, is had good
Good commercial value.There are about 90,000,000 tons of bean dregs in China every year, these bean dregs cannot get reasonable utilization, generates resource wave
Take.If can further investigate to red bacterium bean curd, the product of more excellent kind is createed, will be one considerable
Foundation money road.
But for a long time, Wuping " red bacterium bean curd " more slightly mysterious color, strain is still not clear from He Erlai；
What kind of growth rhythm its main red bacterium of fermentation strain has, and temporarily also there are no people to find out.
Summary of the invention
It is an object of the present invention to provide a kind of Neuraspora crassa strain, the bacterial strain is Neuraspora crassa
(Neurospora crassa) has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation day
Phase is on 01 16th, 2019, and deposit number is CGMCC NO:3.19233.
The present invention passes through traditional " red bacterium bean curd " the main fermentative microflora in separation Wuping, the results showed that neurospora crassa is Wuping
Traditional " red bacterium bean curd " main fermentative microflora, scientific name Neuraspora crassa (Neurosporacrassa) belong to Neurospora
(Neurospora) a kind of Common fungi has branch and separation with loose netted mycelia, and vegetative propagation forms mitogenetic
Spore.Research shows that its nutritional condition is simple, growth is rapid, fermentation period is short, actual production process is easily controllable, and can produce
High activity cellulase, so that the development and utilization for agricultural and sideline waste provide an effectively approach.But traditional " red in Wuping
Bacterium bean curd " fermentation strain is affected mainly by natural " present ", climate, artificial etc., and finished product batch quality is different, therefore work
Industry large-scale production is restricted, and also affects the marketing of the product.In well-known technique, there are some coarse arteries and veins spores at present
Bacterium is applied to the method for the agricultural and sideline products such as straw, stalk production protein feed, but does not have registration also applied to field of food.This
Outside, there are also the methods that report improves Neuraspora crassa fiber conversion capability in well-known technique, but are not applied to food yet
Field.
In view of this, it is another object of the present invention to the production that Neuraspora crassa LY03 is applied to red bacterium bean curd
Cheng Zhong.
To achieve the above object, the invention adopts the following technical scheme: the Neuraspora crassa strain LY03 is seeded to liquid
After fermentation spreads cultivation in body culture medium, the carriers such as gained bacterium solution and corn flour or starch are prepared by mixing into fermentation bacteria cake, by the hair
Red bacterium bean curd is made in yeast-like fungi cake access bean dregs.
Fluid nutrient medium composition are as follows: sucrose 2~3%, corn stalk powder 2.0~3.0%, potassium dihydrogen phosphate 0.1~
0.2%, magnesium sulfate 0.1~0.2%, sodium chloride 0.4~0.5%, pH6.8~7.0.
Fermentation condition are as follows: aerobic, 150~180r/min of revolving speed, 28~30 DEG C, ferment 48~72h.
Further, the red bacterium bean curd the production method is as follows: dried after bean dregs are squeezed, control bean dregs water content exist
Then 20% hereinafter, pave bean dregs, briquetting, the bacteria cake that will ferment uniformly is sprinkling upon bean dregs surface, and ferment 24- at room temperature
48h.In fermentation process, bean dregs surface is covered with to mycelia, by bean dregs turn-over, fermentation bacteria cake is uniformly spread to another surface of bean dregs
Continue to ferment, until another surface of bean dregs covers with mycelia, i.e. fermentation ends, obtains red bacterium bean curd.
The dosage of the fermentation bacteria cake is the 0.05-0.1% of bean dregs quality.
The invention adopts the above technical scheme, provides and a kind of prepares red bacterium bean curd using Neuraspora crassa fermented soybean dregs
Technique.Wherein, the ingredients such as corn flour, the starch added in bacteria cake of fermenting are conducive to the dispersion of fermentation strain mycelia, reinforcing fiber enzyme
Vigor reduces the carbohydrate contents such as fiber, the polysaccharide in bean dregs, increases the content of total protein in finished product, improves mouthfeel.This
For the red bacterium bean curd of invented technology preparation compared with traditional handicraft, total protein, fat content are respectively increased 43.45~61.35%
With 2.08~4.08%, carbohydrate reduces 28.86~29.10%, and cellulase total activity improves 67.27~
95.32%, appearance, mouthfeel are more preferably, it is easier to be favored by consumer.In addition, fermentation bacteria cake preparation process is easy, it is suitble to industry
Metaplasia produces, and the bacterial strain retentive activity time is long, overcomes traditional handicraft fermentation strain by " presenting naturally ", bacterial strain batch is different, matter
Measure disadvantage unstable, that production scale is small.Present invention process can make full use of the waste materials such as bean dregs to carry out industrialized production, open up
Wide soybean and its attached material production industrial chain, have a good application prospect.
Detailed description of the invention
Technical scheme of the present invention will be described in further detail with reference to the accompanying drawings and examples.
Fig. 1 is the flat-plate bacterial colony aspect graph of LY03 bacterial strain；
Fig. 2 is the PCR electrophoretogram in embodiment 1；
Fig. 3 is LY03 bacterial strain affinity figure.
Specific embodiment
Embodiment 1
Separation, the identification of bacterial strain
Using PDA culture medium from Wuping " red bacterium bean curd " of purchase separation and fermentation bacterial strain, and by culture bacterium colony shape
State observation, Microscopic Identification determine the Basic Biological Character of separated fermentation strain, and carry out ITS series primer to the bacterial strain
Sequencing carries out sequence alignment with known bacterial strain on NCBI, determines the species types and affiliation of the bacterial strain.
Concrete technology flow process is as follows:
(1) configuration PDA culture medium: 180~220 grams of potato, 20~25 grams of glucose, 15~20 grams of agar, water 1000
Milliliter, natural PH；Conventionally with postponing, 121 DEG C sterilize 30 minutes, good plate.
(2) separation of bacterial strain
In aseptic operating platform, using aseptic nipper or inoculation shovel dig from the red bacterium bean curd in Wuping of purchase a fritter bacterial strain to
In PDA plate, it is placed in 30 DEG C of incubators and cultivates 24-48 hours.See whether that mycelia grows.
Mycelia: being dug a fritter into new PDA plate by purifying again, is cultivated 24-48 hours again.Mycelia is seen whether
It grows.Mycelia bacterium colony and Anatomical characteristics are observed, plate pollution-free, that mycelia is sturdy is selected and is identified for bacterial strain.
(3) identification of bacterial strain
Selected plate bacterial strain (name LY03) is subjected to bacterium colony and fibre morphology identification, its DAN is extracted, send still raw to platinum
Object technology (Shanghai) Co., Ltd. is sequenced, and is logged in NCBI and is compared, determines its evolutionary relationship and attribute.Flat-plate bacterial colony form is such as
Shown in Fig. 1.The result shows that: the bacterium selected meets the grown form and biological property of fungi, with loose netted bacterium
Silk, mycelia is transparent, colourless or grey, there is branch and separation, and vegetative propagation forms conidium.Ascus size 160~185 × 20
~22 μm, ascospore 8,25~34 × 12~18 μm.Conidium be in light orange, 6~7 μm of diameter.
The analysis of l8S rDNA sequence is carried out to aimed strain LY03.
Forward primer (as shown in SEQ ID NO:1): ITS1:TCCGTAGGTGAACCTGCGG；
Reverse primer (as shown in SEQ ID NO:2): ITS4:TCCTCCGCTTATTGATATGC.Expanding fragment length
600bp or so, PCR electrophoretogram as shown in Fig. 2, measurement primer sequence as shown in SEQ ID NO:3.The primer sequence is logged in
NCBI carries out Blast comparison, and affinity is as shown in Figure 3.The result shows that separated LY03 bacterial strain and Neurospora
(Neurospora) the close source relationship of several bacterial strains is nearest, and for similitude up to 98% or more, the form of comprehensive LY03 bacterial strain is special
Sign and l8S rDNA the sequencing results, are accredited as and are set to Neuraspora crassa (Neurosporacrassa), be commonly called as red
Color bread mold, and bacterial strain is sent to China General Microbiological culture presevation administrative center (CGMCC) preservation, deposit number:
CGMCC3.19233。
Embodiment 2
Using Neuraspora crassa LY03 fermented soybean dregs, red bacterium bean curd is made
(1) production for bacteria cake of fermenting: Neuraspora crassa strain LY03 is seeded in fluid nutrient medium after fermentation spreads cultivation, gained
Bacterium solution is prepared by mixing into fermentation bacteria cake with by dry corn flour, starch (as carrier).
Fluid nutrient medium: sucrose 2.5%, corn stalk powder 2.0%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.1%, chlorine
Change sodium 0.45%, pH6.8.
Fermentation condition: aerobic, revolving speed 150r/min, 28 DEG C, ferment 48h.
(2) production of red bacterium bean curd: the bean dregs of honed soya-bean milk being squeezed, naturally dry, and control bean dregs water content exists
Then 20% hereinafter, pave bean dregs, briquetting, the bacteria cake that will ferment uniformly is sprinkling upon bean dregs surface, and the dosage for bacteria cake of fermenting is beans
The 0.05% of slag amount ferments for 24 hours at room temperature.In fermentation process, bean dregs surface is covered with to mycelia, by bean dregs turn-over,
Fermentation bacteria cake is uniformly spread to another surface of bean dregs to continue to ferment, until another surface of bean dregs covers with mycelia, i.e. fermentation ends,
Obtain red bacterium bean curd.
Measure traditional handicraft Wuping " red bacterium bean curd " of market purchase and " red bacterium bean curd " fermentation of present invention process preparation
The composition of nutriment afterwards, and by the mycelia on two kinds of technique bean dregs surfaces, it accesses in fermentation liquid and ferments (fermentation condition is same as above)
48h, takes supernatant to be used to measure the cellulase total activity of fermentation strain, and measuring method is cellulase total activity measuring method.Comparison
The fermenting property index of two technique, the results are shown in Table 1.Wherein, the measurement of total protein is using Kjeldahl's method, fatty survey
Surely anthrone colorimetry is used using the measurement of soxhlet extraction, carbohydrate.The result shows that " red bacterium manufactured in the present embodiment
For bean curd " compared with traditional handicraft, total protein, fat content are higher, and 43.45% and 2.08%, carbon hydrate has been respectively increased
Object reduces 28.86%, and cellulase total activity improves 67.27%, and appearance, mouthfeel are more preferably.
The red bacterium soybean curd nutrition ingredient comparison of 1 traditional handicraft of table and present invention process preparation
Embodiment 3
Using Neuraspora crassa LY03 fermented soybean dregs, red bacterium bean curd is made
(1) production for bacteria cake of fermenting: Neuraspora crassa strain LY03 is seeded in fluid nutrient medium after fermentation spreads cultivation, gained
Bacterium solution is prepared by mixing into fermentation bacteria cake with by dry corn flour, starch (as carrier).
Fluid nutrient medium: sucrose 3.0%, corn stalk powder 3.0%, potassium dihydrogen phosphate 0.15%, magnesium sulfate 0.15%, chlorine
Change sodium 0.50%, pH7.0.
Fermentation condition: aerobic, revolving speed 180r/min, 30 DEG C, ferment 72h.
(2) production of red bacterium bean curd: the bean dregs of honed soya-bean milk being squeezed, naturally dry, and control bean dregs water content exists
Then 20% hereinafter, pave bean dregs, briquetting, the bacteria cake that will ferment uniformly is sprinkling upon bean dregs surface, and the dosage for bacteria cake of fermenting is beans
The 0.1% of slag amount, ferment 48h at room temperature.In fermentation process, bean dregs surface is covered with to mycelia, it, will by bean dregs turn-over
Fermentation bacteria cake is uniformly spread to another surface of bean dregs and continues to ferment, until another surface of bean dregs covers with mycelia, i.e. fermentation ends, obtains
To red bacterium bean curd.
Measure traditional handicraft Wuping " red bacterium bean curd " of market purchase and " red bacterium bean curd " fermentation of present invention process preparation
The composition of nutriment afterwards, and by the mycelia on two kinds of technique bean dregs surfaces, it accesses in fermentation liquid and ferments (fermentation condition is same as above)
72h, takes supernatant to be used to measure the cellulase total activity of fermentation strain, and measuring method is cellulase total activity measuring method.Comparison
The fermenting property index of two technique, the results are shown in Table 2.Wherein, the measurement of total protein is using Kjeldahl's method, fatty survey
Surely anthrone colorimetry is used using the measurement of soxhlet extraction, carbohydrate.The result shows that " red bacterium manufactured in the present embodiment
For bean curd " compared with traditional handicraft, total protein, fat content are higher, and 72.39% and 4.08%, carbon hydrate has been respectively increased
Object reduces 29.10%, and cellulase total activity improves 95.32%, and appearance, mouthfeel are more preferably.
The red bacterium soybean curd nutrition ingredient comparison of 2 traditional handicraft of table and present invention process preparation
Sequence table
<110>Longyan School
<120>a kind of Neuraspora crassa strain and its application
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tcctccgctt attgatatgc 20
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tcctccgctt attgatatgc ttaagttcag cgggtattcc tacctgatcc gaggtcaacc 60
ttagaaatgg ggggttttac ggcaagaacc cgccgcacga ccatagcgat gtagagttac 120
tacgctcggt gtgactagcg agcccgccac tgattttgag ggaccgcgga cagccgcgga 180
tccccaacgc aagcagagct tgatggttga aatgacgctc gaacaggcat gctcgccaga 240
atactggcga gcgcaatgtg cgttcaaaga ttcgatgatt cactgaattc tgcaattcac 300
attacttatc gcatttcgct gcgttcttca tcgatgccag aaccaagaga tccgttgttg 360
aaagttttga cttatttaaa agtttactca gagagacata aaatatcaag agtttagttt 420
cggcactccg gcgggcagcc tcccgcgagc gggagacccg aggatccggg agggcccgag 480
ggcctttccg gaccgccagc gccgaggcaa ccgtacgggt aagattcgcg atggtttgtg 540
ggagttttgc aactctgtaa tgatccctcc gctggttcac caacggagac cttgttacga 600
cttttacttc c 611
Claims (8)
1. a kind of Neuraspora crassa strain, it is characterised in that: the bacterial strain is Neuraspora crassa Neurospora crassa, has been protected
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, the deposit date is on 01 16th, 2019, preservation
Number be CGMCC NO:3.19233.
2. application of the Neuraspora crassa strain as described in claim 1 in red bacterium bean curd.
3. application according to claim 2, it is characterised in that: the Neuraspora crassa strain LY03 is seeded to Liquid Culture
After fermentation spreads cultivation in base, gained bacterium solution and carrier are prepared by mixing into fermentation bacteria cake, and the fermentation bacteria cake is accessed in bean dregs and is made
Red bacterium bean curd.
4. application according to claim 3, it is characterised in that: the condition to spread cultivation of fermenting is: 28-30 DEG C of temperature, turning
Fast 150-180r/min, aerobic fermentation 48-72h.
5. application according to claim 3, it is characterised in that: the carrier is corn flour or starch.
6. application according to claim 3, it is characterised in that: the red bacterium bean curd the production method is as follows: by bean dregs pressure
It dries, then paves bean dregs, briquetting after squeezing, fermentation bacteria cake is uniformly sprinkling upon bean dregs surface, is sent out at room temperature
Ferment covers with bean dregs surface to mycelia, by bean dregs turn-over, fermentation bacteria cake is uniformly spread to another surface of bean dregs and continues to ferment, directly
Mycelia, i.e. fermentation ends are covered with to another surface of bean dregs, obtains red bacterium bean curd.
7. application according to claim 6, it is characterised in that: the dosage of the fermentation bacteria cake is the 0.05- of bean dregs quality
0.1%。
8. application according to claim 6, it is characterised in that: the control of bean dregs water content after squeezing, drying 20% with
Under.
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Cited By (4)
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|---|---|---|---|---|
| CN109957514A (en) * | 2019-01-25 | 2019-07-02 | 南昌大学 | A kind of new application of sturdy crassa |
| CN114052187A (en) * | 2021-11-16 | 2022-02-18 | 仲恺农业工程学院 | Bean dreg leavening agent, bean dreg fermentation method and fermented bean product |
| CN114606272A (en) * | 2022-03-01 | 2022-06-10 | 河北工程大学 | Preparation method and application of neurospora crassa fermentation extract |
| CN114601092A (en) * | 2022-04-14 | 2022-06-10 | 河北工程大学 | Sweet potato fermented beverage and preparation method thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109957514A (en) * | 2019-01-25 | 2019-07-02 | 南昌大学 | A kind of new application of sturdy crassa |
| CN114052187A (en) * | 2021-11-16 | 2022-02-18 | 仲恺农业工程学院 | Bean dreg leavening agent, bean dreg fermentation method and fermented bean product |
| CN114606272A (en) * | 2022-03-01 | 2022-06-10 | 河北工程大学 | Preparation method and application of neurospora crassa fermentation extract |
| CN114606272B (en) * | 2022-03-01 | 2023-09-26 | 河北工程大学 | Preparation method and application of Neurospora crassa fermentation extract |
| CN114601092A (en) * | 2022-04-14 | 2022-06-10 | 河北工程大学 | Sweet potato fermented beverage and preparation method thereof |
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| Publication number | Publication date |
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| CN110106090B (en) | 2022-12-27 |
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