CN109221878A - A kind of Thallus Porphyrae fermented product and its preparing the application in antioxidant - Google Patents
A kind of Thallus Porphyrae fermented product and its preparing the application in antioxidant Download PDFInfo
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- CN109221878A CN109221878A CN201811372088.4A CN201811372088A CN109221878A CN 109221878 A CN109221878 A CN 109221878A CN 201811372088 A CN201811372088 A CN 201811372088A CN 109221878 A CN109221878 A CN 109221878A
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Abstract
The present invention provides a kind of Thallus Porphyrae fermented product and its preparing the application in antioxidant, the Thallus Porphyrae fermented product the preparation method is as follows: first seaweed is crushed, distilled water is added after sieving；By after activation lactobacillus plantarum or saccharomycete be inoculated with；The obtained Thallus Porphyrae fermented product that ferments is carried out after inoculation.Present invention determine that the best strain of seaweed fermentation is lactobacillus plantarum, the parameter of best zymotechnique are as follows: solid-liquid ratio 1:15, fermentation time 36h, inoculum concentration 3.5%.In the concentration range of measurement, seaweed fermentation liquid has certain reducing power, and has Scavenging activity to DPPH free radical, superoxide anion and hydroxyl radical free radical.Therefore, seaweed can develop good oxidation resistant product.
Description
Technical field
The invention belongs to field of food, and in particular to a kind of Thallus Porphyrae fermented product and its prepare the application in antioxidant.
Background technique
Seaweed (porphyra) alias, purple English, rope dish, sub- dish, film dish, purple beautiful jade, are the general names of marine alternate algae.Seaweed
Belong to marine products red algae, thallus is made of the one layer of cells being embedded in thin layer colloid.Certain seaweed is as a kind of red algae, tool
There is the membranaceous appearance of red algae family.Seaweed type is more, mainly there is Porphyra yezoensis, porphyra haitanensis, laver etc..Seaweed is known as
" ocean vegetables ".Seaweed is considered as important food in oriental countries, soup and the auxiliary material of other meat products can be done, in English
Seaweed is put in and bakes on bread by state, tastes as oyster.Maximum country of consumption is Japan, Korea Spro to seaweed in the world at present
Some countries such as state, China.
Seaweed starts from the artificial cultivation of China in the 1970s, artificial breeding kind is streak Chinese cabbage on a large scale
And porphyra haitanensis.Wherein Porphyra yezoensis is grown in jiangsu coast, the coastal areas such as Nantong, Lianyun Harbour.Porphyra yezoensis harvesting after by
The size that specification is 21cm × 19cm is made, by the Porphyra yezoensis of sheet through overbaking in cleaning, dehydration, chopping, drying, molding
After machine baking, it can be used for making the non-instant sea sedge of sushi, referred to as " non-seasoning sea sedge ".Certain sheet seaweed is added by baking machine
Work, seasoning machine manually add seasoning, then dry, and edible sea sedge is made, referred to as " seasoning sea sedge ".Porphyra haitanensis is grown in
Fujian Coastal Area in Zhejiang Province, growth area ratio Porphyra yezoensis are wider.Seaweed harvest after, through over cleaning, dehydration, drying, molding,
It is sold after packaging, seaweed is generally packaged into round pie, is mainly used for adjusting soup.It has been said that compared with many marine products, seaweed pollution compared with
It is few.Once seawater is contaminated, the growth of seaweed is limited even having no harvest.
The research of the antioxidant activity for concentrating laver amylose, polyphenol, protein etc. main for the research of seaweed at present, with
And the optimization of various composition extraction process.The research of antioxidant activity is upsurge, and the experiment about anti-oxidant research is very more,
More plants with anti-oxidation function are found, and the function of antioxidant is also gradually completed.The antioxidant activity of seaweed
Be possessed by food few in number, and because seaweed yield is big, nutritive value is high, it is cheap the advantages that, receive vast
Consumer's likes.However at present only in some respect for the research of seaweed, as laver amylose, polyphenol, protein are ground
Study carefully, the research that oxidation resistant measurement this respect is integrally carried out for seaweed is also very few.
Summary of the invention
The object of the present invention is to provide a kind of Thallus Porphyrae fermented product and its preparing the application in antioxidant.The present invention is logical
Cross single factor experiment, the measurement research seaweed fermentation of the process optimizations such as orthogonal test and the oxidation resistance to Thallus Porphyrae fermented product
The antioxidant activity of object develops optimal zymotechnique to further prove the high nutritive value of seaweed, for purple in the future
The commercialization research of dish fermentation material provides solid foundation.
For achieving the above object, the present invention is achieved by the following scheme:
The present invention provides a kind of Thallus Porphyrae fermented product, the Thallus Porphyrae fermented product is made by following preparation method:
(1) seaweed is crushed, water is added after sieving, sterilized；
(2) by after activation lactobacillus plantarum or saccharomycete be inoculated with；
(3) it after being inoculated with, ferments at 25-40 DEG C of temperature.
It is further: in the step (2) in 24-72h, clearance rate of the lactobacillus plantarum to DPPH in Thallus Porphyrae fermented product
Greater than saccharomycete.
Further: lactobacillus plantarum optimum inoculation amount is 3% in the step (2).
Further: the best solid-liquid ratio of seaweed is 1: 20 in the step (2).
Further: lactobacillus plantarum fermentation time is 36h in the step (3).
Further: the optimal conditions of fermentation that orthogonal experiment determines in the step (3) is that solid-liquid ratio is 1: 15, inoculum concentration
It is 3.5%, fermentation time 36h.
It is further: to influence three factor primary and secondary sequences of lactobacillus plantarum fermentation seaweed are as follows: solid-liquid ratio > inoculum concentration >
Fermentation time.
The present invention also provides the Thallus Porphyrae fermented products to prepare the application in antioxidant.
Further: the Thallus Porphyrae fermented product has removing energy to DPPH free radical, hydroxyl radical free radical and superoxide anion
Power.
Compared with prior art, advantages of the present invention and have the technical effect that the present invention is determined through experimentation plant cream bar
The best zymotechnique of bacterium fermentation seaweed, the experimental results showed that the antioxidant activity that seaweed tunning has had, is corresponding
The further research of the product development of Thallus Porphyrae fermented product provides theoretical basis.
Detailed description of the invention
Fig. 1 is measurement chart of the different strain fermentation to seaweed DPPH clearance rate；
Fig. 2 is the measurement chart that different feed liquid compares lactobacillus-fermented seaweed；
Fig. 3 is measurement chart of the lactic acid bacteria different vaccination amount to seaweed DPPH clearance rate；
Fig. 4 is influence measurement chart of the different fermentations time for lactobacillus-fermented seaweed；
Fig. 5 is various concentration seaweed concentrate to DPPH free radical scavenging ability measurement chart；
Fig. 6 is Scavenging activity measurement chart of the various concentration seaweed concentrate to superoxide anion；
Fig. 7 is various concentration seaweed fermentation liquid to hydroxyl radical free radical Scavenging activity measurement chart；
Fig. 8 is various concentration seaweed concentrate reducing power measurement chart.
Specific embodiment
Technical solution of the present invention is described in detail below with reference to embodiment.
Embodiment 1: the selection and activation of strain
Lactic acid bacteria (Bulgarian lactobacillus plantarum, commercially available strain) and saccharomycete (wine yeast, city are selected in this experiment
Sell strain) carry out fermentation comparison.
One ring saccharomycete of scraping and lactobacillus plantarum from inclined-plane gently are inoculated into YPD and MRS fluid nutrient medium respectively
On, it for 24 hours in 37 DEG C of cultures, is quantitatively inoculated into seaweed fermentation liquid, passes on 2-3 times, until viable bacteria counts to 1 × 108 or more.
Embodiment 2: measurement of the different strain to DPPH clearance rate in Thallus Porphyrae fermented product
Seaweed is crushed, 100ml distilled water, 121 DEG C of sterilizing 20min are added after crossing 40 meshes.By the plant cream after activation
Bacillus is inoculated with saccharomycete, inoculum concentration 4%, and 37 DEG C of fermentation temperature, fermentation time 48h, revolving speed 120r/min, pH certainly
So.Fermentation ends, fermentation liquid 5000r/min are centrifuged 10min, take supernatant, measure DPPH, and every group setting three parallel.Experiment
As a result as shown in Figure 1.
As shown in Figure 1, saccharomycete and lactobacillus plantarum generally rise the clearance rate of DPPH in Thallus Porphyrae fermented product
Trend.When seaweed fermentation time is during 24-60h, saccharomycete tends to be flat to DPPH free radical scavenging activity in Thallus Porphyrae fermented product
Slow, otherness is not significant (p > 0.05).When seaweed fermentation time is during 24-48h, lactobacillus plantarum is in Thallus Porphyrae fermented product
DPPH free radical scavenging activity tends to ascendant trend, and otherness is significant (p < 0.05), when seaweed fermentation time is during 48-72h,
Lactobacillus plantarum tends towards stability to DPPH free radical scavenging activity in Thallus Porphyrae fermented product, and otherness is not significant (p > 0.05), by Fig. 1
It knows between 24-72h, lactobacillus plantarum is greater than saccharomycete to the clearance rate of DPPH in Thallus Porphyrae fermented product, therefore selects
Lactobacillus plantarum is tested.
Embodiment 3: different feed liquid compares the measurement of DPPH clearance rate in Thallus Porphyrae fermented product
Seaweed 5g, 4g, 3g, 2g, 1g are weighed respectively, and in the 250ml triangular flask equipped with 100 distilled water, material is successively made
The fermentation liquid that liquor ratio is 1: 20,1: 30,1: 40,1: 50,1: 60.Select best strain, inoculum concentration 4%, fermentation temperature 37
DEG C, fermentation time 48h, after fermentation, fermentation liquid carry out centrifuging and taking supernatant and survey DPPH, and every group setting three are parallel.It is real
It is as shown in Figure 2 to test result.
As shown in Figure 2, being gradually reduced with seaweed solid-liquid ratio, under Thallus Porphyrae fermented product is in the clearance rate of DPPH free radical
The trend of drop, when seaweed solid-liquid ratio is 1: 20, DPPH clearance rate is maximum value 86%, is significantly higher than other groups (P < 0.05),
It is that DPPH clearance rate reaches minimum 37% when seaweed solid-liquid ratio is 1: 60.Therefore selecting best solid-liquid ratio is 1: 20.
Embodiment 4: measurement of the different vaccination amount to DPPH clearance rate in Thallus Porphyrae fermented product
Best solid-liquid ratio is selected, seaweed is weighed in the 250ml triangular flask equipped with 100 distilled water, fermentation liquid is made.Inoculation
Amount is set as 2%, 3%, 4%, 5%, 6%, and fermentation temperature is 37 DEG C, fermentation time 48h.After fermentation, fermentation liquid carries out
Centrifuging and taking supernatant surveys DPPH, and every group setting three parallel.Experimental result is as shown in Figure 3.
From the figure 3, it may be seen that being continuously increased with lactobacillus plantarum inoculum concentration, Thallus Porphyrae fermented product is to DPPH clearance rate on first
Downward trend after rising.When lactobacillus plantarum inoculum concentration increases to 3% from 2%, clearance rate constantly rises, when lactobacillus plantarum connects
When kind of amount is 3%, reach maximum value, being worth is 64%, be significantly higher than other groups (P < 0.05), when lactobacillus plantarum inoculum concentration from
3% when increasing to 6%, and Thallus Porphyrae fermented product DPPH clearance rate is in continuous downward trend, therefore when lactobacillus plantarum inoculum concentration is
When 3%, DPPH clearance rate highest, choosing lactobacillus plantarum optimum inoculation amount is 3%.
Embodiment 5: measurement of the different fermentations time to DPPH clearance rate in Thallus Porphyrae fermented product
Best solid-liquid ratio is selected, seaweed is weighed in the 250ml triangular flask equipped with 100 distilled water, fermentation liquid is made.Selection
Best strain, fermentation temperature be 37 DEG C, fermentation time be set as 12h, for 24 hours, 36h, 48h, 60h, after fermentation, fermentation liquid into
Row centrifuging and taking supernatant surveys DPPH, and every group setting three parallel.Experimental result is as shown in Figure 4.
As shown in Figure 4, with the increase of lactobacillus plantarum fermentation time, Thallus Porphyrae fermented product is in first to rise to DPPH clearance rate
The trend to tend towards stability afterwards, during 0-36h, with the increase of seaweed fermentation time, DPPH clearance rate is constantly increased, and works as fermentation
Time is 36h, and DPPH clearance rate is 86%, is significantly higher than other groups (p < 0.05)；During 36-60h, with fermentation time
Increase, DPPH clearance rate tends towards stability, and otherness is not significant (P > 0.05), therefore selects lactobacillus plantarum fermentation time most
Good is 36h.
Embodiment 6: orthogonal experiment
1. lactobacillus-fermented seaweed quadrature factor water-glass of table
Orthogonal Experiment and Design and it the results are shown in Table 2.
2. orthogonal test of table and result
According to single factor experiment as a result, devising Three factors-levels L9 (33) orthogonal test, it investigates result and utilizes
DPPH clearance rate indicates, it is thus found that three factor primary and secondarys sequences for influencing lactobacillus plantarum fermentation seaweed are A > B > respectively
C, i.e. solid-liquid ratio > inoculum concentration > fermentation time；The optimum process condition of extraction is A1B3C2, i.e., solid-liquid ratio is 1: 15 (g/ml),
Inoculum concentration is 3.5%, fermentation time 36h, carries out the measurement of DPPH clearance rate, clearance rate according to orthogonal test optimum process condition
It is 92%, is higher than above 9 combinations, orthogonal test optimised process is verified.
Embodiment 7: the measurement of seaweed in vitro anti-oxidation
The Thallus Porphyrae fermented product of 200ml optimised process is taken to dilute 5 times, 6000r/min is centrifuged 20 minutes, and centrifugate is revolved
Inspissation contracting, resulting concentrate is bottled, for 24 hours in -24 DEG C of pre-freezes, the good concentrate of pre-freeze is put into freeze dryer, is freezed
Dry 72h, obtains seaweed condensed powders, and weigh to obtain 5mg, dilution.
1, the measurement of Thallus Porphyrae fermented product scavenging ability of DPPH free radical
DPPH determining free radicals are shown in Table 3 using DPPH method.
The method of 3. scavenging ability of DPPH free radical of table
Note: -- any reagent is not added in representative.
Laboratory is equipped with blank tube and sample cell, the DPPH of 2.4ml 0.2mM will be separately added into control tube and sample cell,
The dehydrated alcohol of 2.4ml is added into sample ginseng pipe again, extraction examination is added in blank tube, control tube, sample cell, sample ginseng pipe respectively
Then 0.5ml sample is added in sample cell and sample ginseng pipe, in room temperature after mixing well in agent 4ml, 1.6ml, 1.1ml, 1.1ml
Dim place stands 30min, and absorbance is surveyed at 517nm.
Obtained sample is made into the solution of various concentration, measures the clear ability of its DPPH free radical, as a result such as Fig. 5 institute
Show.As shown in Figure 5, with the increase of Thallus Porphyrae fermented product mass concentration, fermentation material is after first increasing to DPPH free radical scavenging activity
Tend towards stability, during concentration 0-4mg/ml, Thallus Porphyrae fermented product be to the clearance rate of DPPH free radical it is in rising trend, it is poor
Anisotropic significant (p < 0.05)；During concentration 4-10mg/ml, Thallus Porphyrae fermented product tends towards stability to the clearance rate of DPPH free radical,
There was no significant difference (p > 0.05).It follows that Thallus Porphyrae fermented product has certain Scavenging activity for DPPH free radical, and
Increase with the increase of mass concentration.
2, Thallus Porphyrae fermented product removes the measurement of superoxide anion ability
The removing of superoxide anion uses assay NBT photoreduction, is shown in Table 4.
Table 4. removes the active method of ultra-oxygen anion free radical
Note: -- represent not reagent adding.
Four test tubes are taken respectively, 5mlTris-HCl is added in every test tube, and the extraction of 50ul is added into 1, No. 2 test tube
The sample of 50ul is added in 3, No. 4 test tubes, first mixes test tube for reagent, will add in 1,4 test tubes after 25 DEG C of standings water-bath 20min
Enter 10mM hydrochloric acid, the 25mM pyrogallol of 40u1, test tube is mixed in 2, No. 3 test tubes, is added dropwise rapidly after 25 DEG C of standing 3min
50ul DTT, is stored at room temperature 15min, and OD value is measured at 316nm.
It will be appreciated from fig. 6 that Thallus Porphyrae fermented product is to superoxide anion Scavenging activity with the increase of Thallus Porphyrae fermented product mass concentration
Be it is in rising trend, Thallus Porphyrae fermented product mass concentration be 1-5mg/ml during, Thallus Porphyrae fermented product is for the clear of superoxide anion
Except there was no significant difference (p > 0.05) for rate, when Thallus Porphyrae fermented product mass concentration is in 5mg/ml, clearance rate reaches maximum, most
Big value is 30%, thus, Thallus Porphyrae fermented product has certain elimination effect to superoxide anion, and with the increase of concentration
And it is gradually increasing.
3, Thallus Porphyrae fermented product removes the measurement of hydroxyl radical free radical ability
The measurement of hydroxy radical (OH) uses o-phenanthroline, is shown in Table 5.
Table 5. removes the active measuring method of hydroxyl radical free radical
Note: -- any reagent is not added in representative.
3.25ml PBS is added in every test tube, and 0.75ml distilled water, backward No. 2 test tubes are then added into 1, No. 4 test tube
Middle addition 0.25ml distilled water；Then 1ml sample is added into 4, No. 5 test tubes, 1ml extraction is added in backward 1,2, No. 3 test tube
0.25ml Phen, the test tube rotated after mixing well on vortex mixer (2,3,5) are added into 2,3, No. 5 test tubes for reagent
The middle ferric sulfate that 0.25ml is added, then into 3, No. 5 test tubes, the hydrogen peroxide of addition 0.25ml is put into 37 DEG C of water-baths, is stood
Absorbance is surveyed after 90min at 536nm.
Obtained sample is made into the solution of various concentration, measures the Scavenging activity of its hydroxyl radical free radical, as a result such as Fig. 7 institute
Show.As shown in Figure 7, with the increase of mass concentration, fermentation material be for the clearance rate of hydroxyl radical free radical it is in rising trend, when
For mass concentration during 1-3mg/ml, Thallus Porphyrae fermented product tends towards stability to hydroxyl radical free radical clearance rate, there was no significant difference (p >
0.05), in 3-5mg/ml, Thallus Porphyrae fermented product is in rising trend to hydroxyl radical free radical clearance rate, significant (the p < of otherness
0.05), when mass concentration is 5mg/ml, Thallus Porphyrae fermented product is maximum to the clearance rate of hydroxyl radical free radical, maximum value 30%, by
This is it is found that Thallus Porphyrae fermented product has scavenging effect to hydroxyl radical free radical in certain concentration range, and with concentration
Increase and increases.
Embodiment 8: the measurement of the total reducing power of Thallus Porphyrae fermented product
The measurement of reducing power uses potassium ferricyanide reduction method, is shown in Table 6.
6. reducing power measuring method of table
Note: -- any reagent is not added in representative.
Prepare blank tube and sample cell first, 1ml distilled water is added in blank tube, 1ml sample is added in sample cell, then
Into each test tube be added 0.2mol/L PBS (PH=6.6) 0.2ml, 1% potassium ferricyanide solution 0.5ml, after mixing well in
20min is placed under the conditions of 50 DEG C, 10%TCA1ml is added after flowing water is cooling, after mixing, in 5000r/min, is centrifuged 20min, is taken
1% iron chloride 0.2ml and distilled water 3ml is added in 1.5ml supernatant, shakes up and surveys absorbance after standing 5min at 720nm, inhales
Light value size and reducing power size are directly proportional.
As shown in Figure 8, with the increase of Thallus Porphyrae fermented product mass concentration, Thallus Porphyrae fermented product reducing power is in rising trend.
For mass concentration during 100-500ug/ml, otherness is significant (p < 0.05), when concentration is 500ug/ml, Thallus Porphyrae fermented product
Reducing power reach maximum, maximum value 0.06, it follows that reducing power of the Thallus Porphyrae fermented product in certain concentration range
Be it is stronger, reducing power also with mass concentration increase and increase.
The result shows that Thallus Porphyrae fermented product to DPPH free radical, hydroxyl radical free radical, reducing power, superoxide anion clearance rate most
A height of 69%, 30%, 0.06,30%.It follows that seaweed fermentation liquid has certain also proper energy in the concentration range of measurement
Power, and there is Scavenging activity to DPPH free radical, superoxide anion and hydroxyl radical free radical.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than is limited；Although referring to aforementioned reality
Applying example, invention is explained in detail, for those of ordinary skill in the art, still can be to aforementioned implementation
Technical solution documented by example is modified or equivalent replacement of some of the technical features；And these are modified or replace
It changes, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (9)
1. a kind of Thallus Porphyrae fermented product, it is characterised in that the Thallus Porphyrae fermented product is made by following preparation method:
(1) seaweed is crushed, water is added after sieving, sterilized；
(2) by after activation lactobacillus plantarum or saccharomycete be inoculated with；
(3) it after being inoculated with, ferments at 25-40 DEG C of temperature.
2. Thallus Porphyrae fermented product according to claim 1, it is characterised in that: in the step (2) in 24-72h, plant cream
Bacillus is greater than saccharomycete to the clearance rate of DPPH in Thallus Porphyrae fermented product.
3. Thallus Porphyrae fermented product according to claim 1, it is characterised in that: lactobacillus plantarum most preferably connects in the step (2)
Kind amount is 3%.
4. Thallus Porphyrae fermented product according to claim 1, it is characterised in that: the best solid-liquid ratio of seaweed is in the step (2)
1:20。
5. Thallus Porphyrae fermented product according to claim 1, it is characterised in that: when lactobacillus plantarum ferments in the step (3)
Between be 36h.
6. Thallus Porphyrae fermented product according to claim 1, it is characterised in that: orthogonal experiment determines most in the step (3)
Good fermentation condition is that solid-liquid ratio is 1:15, inoculum concentration 3.5%, fermentation time 36h.
7. Thallus Porphyrae fermented product according to claim 1, it is characterised in that: influence lactobacillus plantarum fermentation three of seaweed because
Plain primary and secondary sequence are as follows: solid-liquid ratio > inoculum concentration > fermentation time.
8. Thallus Porphyrae fermented product according to claim 1 is preparing the application in antioxidant.
9. Thallus Porphyrae fermented product according to claim 9 is preparing the application in antioxidant, it is characterised in that: the seaweed
Fermentation material has Scavenging activity to DPPH free radical, hydroxyl radical free radical and superoxide anion.
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| CN113174411A (en) * | 2021-03-16 | 2021-07-27 | 集美大学 | Lactobacillus and rhodophyta fermentation supernatant with alpha-glucosidase inhibitory activity and application thereof |
| CN113383917A (en) * | 2021-06-28 | 2021-09-14 | 集美大学 | Preparation method of red algae-lactobacillus fermentation liquor for inhibiting lipase activity |
| CN115386509A (en) * | 2022-07-11 | 2022-11-25 | 江苏海洋大学 | Lactobacillus plantarum MMB-05 and application thereof in preparation of laver sauce by fermenting seaweed leftovers |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113174411A (en) * | 2021-03-16 | 2021-07-27 | 集美大学 | Lactobacillus and rhodophyta fermentation supernatant with alpha-glucosidase inhibitory activity and application thereof |
| CN113383917A (en) * | 2021-06-28 | 2021-09-14 | 集美大学 | Preparation method of red algae-lactobacillus fermentation liquor for inhibiting lipase activity |
| CN115386509A (en) * | 2022-07-11 | 2022-11-25 | 江苏海洋大学 | Lactobacillus plantarum MMB-05 and application thereof in preparation of laver sauce by fermenting seaweed leftovers |
| CN115386509B (en) * | 2022-07-11 | 2023-12-29 | 江苏海洋大学 | Lactobacillus plantarum MMB05 and application thereof in preparing laver sauce from fermented sea weed leftovers |
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