CN106635850B - A kind of marine animal homogenate removing heavy-metal method - Google Patents
A kind of marine animal homogenate removing heavy-metal method Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 43
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- 239000006228 supernatant Substances 0.000 claims abstract description 46
- 241000894006 Bacteria Species 0.000 claims abstract description 42
- 241000235342 Saccharomycetes Species 0.000 claims abstract description 28
- 238000001556 precipitation Methods 0.000 claims abstract description 17
- 238000002360 preparation method Methods 0.000 claims abstract description 17
- 238000012545 processing Methods 0.000 claims abstract description 10
- 238000000926 separation method Methods 0.000 claims abstract description 9
- 230000001376 precipitating effect Effects 0.000 claims abstract description 8
- 239000002244 precipitate Substances 0.000 claims abstract description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 71
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 46
- 239000007788 liquid Substances 0.000 claims description 27
- 239000011780 sodium chloride Substances 0.000 claims description 23
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 20
- 230000004913 activation Effects 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 13
- 239000002253 acid Substances 0.000 claims description 13
- 235000010413 sodium alginate Nutrition 0.000 claims description 13
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- 241000235648 Pichia Species 0.000 claims description 9
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 4
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- 206010018910 Haemolysis Diseases 0.000 claims description 3
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- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 2
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- 238000005469 granulation Methods 0.000 claims 1
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- 229910052785 arsenic Inorganic materials 0.000 description 3
- RQNWIZPPADIBDY-UHFFFAOYSA-N arsenic atom Chemical compound [As] RQNWIZPPADIBDY-UHFFFAOYSA-N 0.000 description 3
- 239000001110 calcium chloride Substances 0.000 description 3
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- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
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- 241000196324 Embryophyta Species 0.000 description 1
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- 238000002525 ultrasonication Methods 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
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- C12N1/14—Fungi; Culture media therefor
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- C12N1/18—Baker's yeast; Brewer's yeast
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Abstract
The present invention provides a kind of marine animal homogenate removing heavy-metal methods, saccharomycete is expanded culture first, prepare fixed yeast gel ball, and preparation marine animal homogenate, acidolysis marine animal homogenate simultaneously carries out isolated supernatant and bottom precipitation, finally using the heavy metal in yeast gel ball removing bottom precipitation, and utilize the heavy metal in free yeast bacterium absorption supernatant.The present invention is directed to the homogenate of marine organisms, it is Promethean propose by homogenate centrifugation be divided into supernatant precipitate two parts processing, supernatant carries out removing heavy-metal processing using yeast bacterium mud, precipitating is removed using fixed yeast gel ball, and the removal efficiency of heavy metal can be significantly improved compared with whole homogenates.Microorganism is immobilized processing by the present invention, can not only have been played microorganism to the absorption property of heavy metal but also can preferably have been realized the separation of microorganism and food material, and industrialized utilization is convenient for.
Description
Technical field
The present invention relates to a kind of marine animal homogenate removing heavy-metal methods, belong to technical field of food biotechnology.
Background technique
With the development of industry with the increase of mankind's activity, the heavy metal pollution in the environment such as big gas and water, soil is increasingly tight
Weight, wherein Heavy Metal Pollution in Water Environment of China problem is very prominent.Heavy metal pollution in water environment suffers aquatic ecosystem
To different degrees of destruction, many aquatiles including fish, shrimp, shellfish are with higher to the heavy metal in its growing environment
Enrichment causes heavy metal pollution event in aquatic products to happen occasionally, and heavy metal pollution, which has become, influences Safety of Aquatic Products
One of an important factor for.Heavy metal can not degrade in the environment, and can accumulate in vivo, and continuous by food chain
Enrichment and transmitting, finally enter human body.Heavy metal is to endanger one of maximum pollutant to the mankind, and toxicological effect mainly shows
To cause the acute and chronic injury of body, causing dysgenesia, hinder embryo's normal development etc..
At present from aquatic products remove heavy metal method mainly have living body temporary feeding purification, chitosan method, chelating resin method,
Complexometry, absorption method etc., these methods some there is at high cost, energy consumption is high, operating process is cumbersome, some easily cause secondary dirt
The disadvantages of dye, especially when concentration of heavy metal ion is slightly exceeded, often due to operating cost is excessively high and is difficult to promote to answer
With.
Method using the absorption of microorganism and absorption removing heavy metal is called microbial method, research shows that some can
Edible saccharomycete, bacterium, mould etc. also has preferable heavy metal removing effect.Microbial method removing heavy-metal has source rich
It is rich, at low cost, safe and non-toxic, treatment effeciency is high, be suitble to the advantages that applying in complicated food material.But microbial method
There are scavengers to be difficult to isolated problem with removing feed liquid.
Summary of the invention
Safe and non-toxic the object of the present invention is to provide a kind of marine animal homogenate removing heavy-metal method, effectively removing is extra large
Heavy metal in foreign animal homogenate, and can be realized industrial application, to make up the deficiencies in the prior art.
The present invention specifically utilizes the absorption of microorganism and absorption function to take off heavy metal ion from animal sources food material
It removes, and microorganism is immobilized into Gel Treatment, and can quickly and effectively be separated with homogenate after removing heavy metal.
In order to achieve the above objectives, the specific technical solution that the present invention takes is:
A kind of marine animal homogenate removing heavy-metal method, first expands culture saccharomycete, prepares immobilization ferment
Female gel ball, and preparation marine animal homogenate, acidolysis marine animal homogenate simultaneously carry out isolated supernatant and bottom
Precipitating, finally using the heavy metal in yeast gel ball removing bottom precipitation, and using in free yeast bacterium absorption supernatant
Heavy metal.
Further, the saccharomycete includes the one of Pichia kudriavezii, saccharomyces cerevisiae, Lu Shi yeast, brewer's yeast etc.
Kind or a variety of mixing.
Further, osmotic pressure Stress treatment is carried out to saccharomycete during above-mentioned saccharomycete expands culture.
Further, the osmotic pressure stress conditions include NaCl Stress or/and carbohydrate stress.
Further, the carbohydrate stress is preferably sucrose stress.
Further, the acid solution in the acid hemolysis process includes one or more mixing such as hydrochloric acid, phosphoric acid, lactic acid.
There are the cross protection phenomenon between stress conditions in microorganism, i.e., a kind of stress conditions can help microorganism to resist
The harm of other stress conditions.In eukaryotic microorganisms, the stress conditions in growing environment can induce cell and generate to other differences
The cross protection of stress conditions.Carrying out infiltration pressure stress to eukaryotic microorganisms can be improved its resistance to heavy metals such as cadmium, zinc.
The removing heavy-metal method specific implementation step includes:
1) saccharomycete activation and the preparation of yeast paste
Saccharomycete carries out solid activation first, i.e., saccharomycete is inoculated in solid slope culture medium, in constant incubator
Culture；Liquid activation is then carried out, the saccharomycete of slant activation is inoculated in fluid nutrient medium, shaken cultivation；It takes again certain
The yeast bacteria suspension of volume of liquid activation is added in the fluid nutrient medium of sodium chloride-containing and/or sugar, shaken cultivation；Culture terminates
Afterwards, fermentation liquid centrifugation, flocculation or film are filtered, yeast bacterium mud is obtained after cleaning；
2) preparation of fixed yeast gel ball
Sodium alginate is taken to dissolve in aqueous solution, ultrasonic degassing processing prepares sodium alginate soln；Ferment prepared by step 1)
Female bacterium mud and sodium alginate after mixing, prepare fixed yeast gel ball using granulating technique, gel media is preferably chlorine
Change calcium solution, standing, sterile water or aseptic sodium chloride solution or calcium chloride cleaning, it is standby to obtain fixed yeast gel ball
With；
3) the acidolysis processing of marine animal homogenate
It is homogenized after marine animal is pre-processed according to a conventional method, obtains marine animal homogenate；It is added pH0.5~pH6's
The quality volume (g/ml) of acid solution stirring or oscillation, marine animal homogenate and acid solution is than being 1:1~1:20, reaction knot
Shu Hou, then stand, it is then centrifuged for or film filters, collect supernatant and bottom precipitation respectively；
4) in marine animal homogenate supernatant heavy metal removing
The yeast bacterium mud that step 1) is obtained is added in the marine animal homogenate supernatant of step 3) preparation, adjusts supernatant
The pH of liquid is 2~10,10~35 DEG C, perhaps 5~240min of stirring centrifugation or flocculation or film are filtered to remove yeast for oscillation
Bacterium, the supernatant after obtaining removing heavy metal；
5) in marine animal homogenate bottom precipitation heavy metal removing
It is heavy that fixed yeast gel ball prepared by step 2) is added to the marine animal homogenate bottom that step 3) obtains
It in shallow lake, adds water and stirs, adjusting pH is 2~10,10~35 DEG C, 0.2~8h of shaken cultivation, and collection fixed yeast gel ball is simultaneously clear
It washes, the nutriments such as the albumen in liquid and gel ball cleaning solution is removed using isoelectric point method or flocculence recycling.
Further, cleaning yeast bacterium mud selects sterile water or aseptic sodium chloride solution in the step 1).
Further, in the step 1) sodium chloride concentration be 0.1wt%~40wt%, sugared concentration be 0.1wt%~
50wt%；The concentration of sodium chloride is preferably 8wt%.
Further, the concentration of sodium alginate is 0.5wt%~5wt% in the step 2), and calcium chloride concentration is
0.1wt%~6wt%.
Further, acid solution is preferably hydrochloric acid in the step 3), and pH is preferably 3.
Further, the pH removed in the step 4) and step 5) is preferably 8.
The invention has the advantages that the present invention is directed to the characteristics of homogenate of marine organisms is in sauce shape poor fluidity, originality
Propose homogenate is handled after be centrifuged, be divided into supernatant precipitate two parts processing, supernatant using yeast bacterium mud into
The processing of row removing heavy-metal, precipitating are removed using fixed yeast gel ball.This method can either effectively remove heavy metal, again
Can be convenient for filtering and removing and recycling, the removing of heavy metal suitable for for sauce shape poor fluidity food material, and
The effect for dividing supernatant and bottom precipitation two parts respectively to be removed is more preferable than the removal effect of complete homogenate, removing knot
Shu Hou, dry matter yield is higher, largely avoids the loss of nutriment.
In addition, homogenate is carried out acidolysis and extracts heavy metal in acid condition by the present invention, it is easy to operate at low cost,
Removal efficiency is high；The present invention carries out Stress treatment in saccharomycete incubation, can significantly increase the heavy metal removing of saccharomycete
Efficiency.Yeast is separated from certain food, safe and non-toxic, is that one plant of salt resistance, antiacid, impermeabilisation pressure, resistant to high temperatures etc. are more
Resistant yeast, while there is various heavy adsorption capacity, heavy metal ion can be adsorbed in complicated food material；And this hair
Bright selection Pichia kudriavezii, the good removal effect of heavy metal is belonged to present invention firstly discovers that.What the present invention used
Sodium alginate and calcium chloride are food-grade, safe and non-toxic, it can be ensured that foodsafety.The present invention immobilizes microorganism
Processing, can not only play microorganism to the absorption property of heavy metal but also can preferably realize the separation of microorganism and food material, just
In industrialized utilization.The present invention has the characteristics that easy to operate, safe and non-toxic, at low cost and is conducive to industrial applications.
Detailed description of the invention
Fig. 1 is process flow chart of the invention；
Fig. 2 is fixed yeast gel ball schematic diagram produced by the present invention；
Fig. 3 is concentration of hydrochloric acid in marine animal homogenate acid hemolysis process to homogenate heavy metal removing effect picture；
Fig. 4 is Pichia kudriavezii sodium chloride concentration and pH to homogenate heavy metal removing effect picture.
Specific embodiment
The invention will be described in further detail with reference to embodiments.
Embodiment 1
Removing heavy-metal method in oyster homogenate, the specific steps are as follows:
1) culture of Pichia kudriavezii and the preparation of yeast bacterium mud
Slant activation: the strain inoculated of 4 DEG C of preservations to YPD slant medium is cultivated for 24 hours in 28 DEG C of constant incubators；
Fluid nutrient medium activation: the thallus after oese scraping slant activation is inoculated on a small quantity in YPD fluid nutrient medium, permanent
28 DEG C in warm shaken cultivation case, 180r/min is cultivated for 24 hours, is seed liquor；
NaCl Stress culture: taking activated saccharomycete seed liquor 1ml, is inoculated into the YPD liquid training containing 8% sodium chloride
28 DEG C, 180r/min culture 20h are supported in base.
Collect bacterium mud: quantitative using blood counting chamber after NaCl Stress culture 20h.Take the saccharomycete of certain volume outstanding
Liquid centrifugation abandons supernatant and it is spare twice with sterile water wash to obtain bacterium mud.
2) acidolysis oyster homogenate
Fresh oyster decladding is homogenized 5min, obtains oyster homogenate.The hydrochloric acid solution of oyster homogenate and pH3 are pressed into matter
Amount is mixed with the solid-liquid ratio of volume 1:5, oscillation or ultrasonication 1h at 25 DEG C.It stands or is centrifuged, collect supernatant respectively
Liquid and bottom precipitation.It takes supernatant to carry out wet digestion, surveys cadmium content with ICP-MS.
3) cadmium in supernatant is taken off
The supernatant for taking acidolysis oyster homogenate and obtaining after being centrifuged, the pH for adjusting supernatant is no more than 8, by 1.3*108
Yeast bacterium mud is added in the additive amount of a/ml.28 DEG C, 180r/min processing 2h.It is centrifuged after absorption, takes supernatant and substrate heavy
It forms sediment and carries out wet digestion, survey cadmium content with ICP-MS.
It takes the oyster homogenate of certain mass to carry out wet digestion, cadmium content is surveyed with ICP-MS, to calculate removal efficiency.
Oyster homogenate is after acidolysis is centrifuged, and cadmium content reaches standard requirements in substrate precipitating, need to only continue to use ferment at this time
Female bacterium mud takes off the cadmium in supernatant, and removal efficiency 53.05%, cadmium concentration is 1.039mg/kg after removing, meets the requirements of the standard.
As a result as shown in table 1 below:
1 oyster homogenate of table is by comparison before and after acidolysis-yeast adsorbing and removing cadmium
| Project | Before removing | After removing | Removal efficiency |
| Cadmium (mg/kg) | 2.213 | 1.039 | 53.05% |
Embodiment 2
The method of removing heavy-metal in sea cucumber intestine homogenate, specific step is as follows (as shown in Figure 1):
1) culture of Pichia kudriavezii and the preparation of yeast bacterium mud
Slant activation: the strain inoculated of 4 DEG C of preservations to YPD slant medium is cultivated for 24 hours in 28 DEG C of constant incubators；
Fluid nutrient medium activation: the thallus after oese scraping slant activation is inoculated on a small quantity in YPD fluid nutrient medium, permanent
28 DEG C in warm shaken cultivation case, 180r/min is cultivated for 24 hours, is seed liquor；
NaCl Stress culture: taking activated saccharomycete seed liquor 1ml, is inoculated into the YPD liquid training containing 8% sodium chloride
28 DEG C, 180r/min culture 20h are supported in base.
Collect bacterium mud: quantitative using blood counting chamber after NaCl Stress culture 20h.Take the saccharomycete of certain volume outstanding
Liquid centrifugation abandons supernatant and it is spare twice with sterile water wash to obtain bacterium mud.
2) preparation of fixed yeast gel ball
The yeast bacterium mud that step 1) obtains is uniformly mixed with 4% sodium alginate soln, is added drop-wise to 1% calcium chloride solution
In, the volume ratio of yeast bacteria suspension and sodium alginate is 4:1.After standing 20min is fixed, sterile water wash 2~3 times, consolidate
Surely change yeast gel ball, as shown in Figure 2.
3) acidolysis sea cucumber intestine homogenate
Sea cucumber intestine goes sand to clean, and is homogenized 5min, obtains sea cucumber intestine homogenate.The hydrochloric acid of sea cucumber intestine homogenate and pH3 is molten
Liquid is mixed by quality with the solid-liquid ratio of volume 1:5, oscillation treatment 20min at 25 DEG C.Centrifugation, collects supernatant respectively and bottom is heavy
It forms sediment.Supernatant and bottom precipitation is taken to carry out wet digestion, with ICP-MS check weighing tenor.
4) heavy metal in supernatant is taken off
The supernatant for taking acidolysis sea cucumber intestine homogenate and obtaining after being centrifuged, the pH for adjusting supernatant is no more than 8, by 2.5*
108Yeast bacterium mud is added in the additive amount of a/ml.28 DEG C, 180r/min oscillation treatment 30min.It is centrifuged after absorption, takes supernatant
Liquid carries out wet digestion, with ICP-MS check weighing tenor.If supernatant content of beary metal is not up to standard, it is straight to continue above-mentioned steps
It is extremely up to standard.If heavy metals exceeding standard in bottom precipitation, enter next step.
5) in bottom precipitation heavy metal removing
Obtained bottom precipitation after taking acidolysis sea cucumber intestine homogenate and being centrifuged, is added appropriate ultrapure water and adjusts pH and be no more than
8, the fixed yeast gel ball of step 2) preparation is added, is added in every 1g substrate precipitating fixed made of 15ml yeast bacteria suspension
Change yeast gel ball.28 DEG C, 180r/min absorption 2h.After absorption, using the method for filtering by fixed yeast gel ball
With feed separation, and wet digestion is carried out to it, with ICP-MS check weighing tenor.
It takes the sea cucumber intestine homogenate of certain mass to carry out wet digestion, with ICP-MS check weighing tenor, is taken off to calculate
Except rate.
The removal efficiency of arsenic is 61.93% in sea cucumber intestine, and the removal efficiency of cadmium is 52.63%, and the nutriment rate of recovery is 89%,
Greatly avoid the loss of nutriment.
As a result as shown in table 2 below:
2 sea cucumber intestine homogenate of table is by comparison before and after acidolysis-yeast adsorbing and removing heavy metal
Embodiment 3
The method of removing heavy-metal in sea cucumber ovum homogenate, the specific steps are as follows:
1) culture of Pichia kudriavezii and the preparation of yeast bacterium mud
Slant activation: the strain inoculated of 4 DEG C of preservations to YPD slant medium is cultivated for 24 hours in 28 DEG C of constant incubators；
Fluid nutrient medium activation: the thallus after oese scraping slant activation is inoculated on a small quantity in YPD fluid nutrient medium, permanent
28 DEG C in warm shaken cultivation case, 180r/min is cultivated for 24 hours, is seed liquor；
NaCl Stress culture: taking activated saccharomycete seed liquor 1ml, is inoculated into the YPD liquid training containing 8% sodium chloride
28 DEG C, 180r/min culture 20h are supported in base.
Collect bacterium mud: quantitative using blood counting chamber after NaCl Stress culture 20h.Take the saccharomycete of certain volume outstanding
Liquid centrifugation abandons supernatant and it is spare twice with sterile water wash to obtain bacterium mud.
2) preparation of fixed yeast gel ball
The yeast bacterium mud that step 1) obtains is uniformly mixed with 2% sodium alginate soln, is added drop-wise to 3% calcium chloride solution
In, the volume ratio of yeast bacteria suspension and sodium alginate is 4:1.After standing 20min is fixed, sterile water wash 2~3 times, consolidate
Surely change yeast gel ball.
3) acidolysis sea cucumber ovum homogenate
Sea cucumber ovum goes sand to clean, and is homogenized 5min, obtains sea cucumber ovum homogenate.The hydrochloric acid solution of sea cucumber ovum homogenate and pH3
It is mixed by quality with the solid-liquid ratio of volume 1:5, oscillation treatment 20min at 25 DEG C.Centrifugation, collects supernatant respectively and bottom is heavy
It forms sediment.Supernatant and bottom precipitation is taken to carry out wet digestion, with ICP-MS check weighing tenor.
4) heavy metal in supernatant is taken off
The supernatant for taking acidolysis sea cucumber ovum homogenate and obtaining after being centrifuged, the pH for adjusting supernatant is no more than 8, by 2.5*
108Yeast bacterium mud is added in the additive amount of a/ml.28 DEG C, 180r/min oscillation treatment 30min.It is centrifuged after absorption, takes supernatant
Liquid carries out wet digestion, with ICP-MS check weighing tenor.
5) in bottom precipitation heavy metal removing
Obtained bottom precipitation after taking acidolysis sea cucumber ovum homogenate and being centrifuged, is added appropriate ultrapure water and adjusts pH and be no more than
8, the fixed yeast gel ball of step 2) preparation is added, is added in every 1g substrate precipitating fixed made of 15ml yeast bacteria suspension
Change yeast gel ball.28 DEG C, 180r/min absorption 2h.After absorption, using the method for filtering by fixed yeast gel ball
With feed separation, and wet digestion is carried out to it, with ICP-MS check weighing tenor.
It takes the sea cucumber ovum homogenate of certain mass to carry out wet digestion, with ICP-MS check weighing tenor, is taken off to calculate
Except rate.
As a result as shown in table 3 below:
3 sea cucumber ovum homogenate of table is by comparison before and after acidolysis-yeast adsorbing and removing heavy metal
| Project | Before removing | After removing | Removal efficiency |
| Arsenic (mg/kg) | 2.727 | 0.678 | 75.14% |
Comparative example 1:
The method of removing heavy-metal in sea cucumber ovum homogenate, the specific steps are as follows:
1) culture of Pichia kudriavezii and the preparation of yeast bacterium mud
Slant activation: the strain inoculated of 4 DEG C of preservations to YPD slant medium is cultivated for 24 hours in 28 DEG C of constant incubators；
Fluid nutrient medium activation: the thallus after oese scraping slant activation is inoculated on a small quantity in YPD fluid nutrient medium, permanent
28 DEG C in warm shaken cultivation case, 180r/min is cultivated for 24 hours, is seed liquor；
NaCl Stress culture: taking activated saccharomycete seed liquor 1ml, is inoculated into the YPD liquid training containing 8% sodium chloride
28 DEG C, 180r/min culture 20h are supported in base.
Collect bacterium mud: quantitative using blood counting chamber after NaCl Stress culture 20h.Take the saccharomycete of certain volume outstanding
Liquid centrifugation abandons supernatant and it is spare twice with sterile water wash to obtain bacterium mud.
2) preparation of fixed yeast gel ball
The yeast bacterium mud that step 1) obtains is uniformly mixed with 2% sodium alginate soln, is added drop-wise to 3% calcium chloride solution
In, the volume ratio of yeast bacteria suspension and sodium alginate is 4:1.After standing 20min is fixed, sterile water wash 2~3 times, consolidate
Surely change yeast gel ball.
3) acidolysis sea cucumber ovum homogenate
Sea cucumber ovum goes sand to clean, and is homogenized 5min, obtains sea cucumber ovum homogenate.The hydrochloric acid of sea cucumber ovum homogenate and pH3 is molten
Liquid is mixed by quality with the solid-liquid ratio of volume 1:5, oscillation treatment 20min at 25 DEG C.
4) heavy metal in acid hydrolysis solution is taken off
The sea cucumber ovum homogenate for taking acidolysis to obtain is added suitable ultrapure water and adjusts pH no more than 8, step 2) system is added
Fixed yeast gel ball made of 15ml yeast bacteria suspension is added in every 10g acid hydrolysis solution for standby fixed yeast gel ball.28
DEG C, 180r/min adsorb 2h.After absorption, using the method for filtering by fixed yeast gel ball and feed separation, and it is right
It carries out wet digestion, with ICP-MS check weighing tenor.
It takes the sea cucumber ovum homogenate of certain mass to carry out wet digestion, with ICP-MS check weighing tenor, is taken off to calculate
Except rate.
As a result as shown in table 5 below:
5 sea cucumber ovum homogenate of table is by comparison before and after acidolysis-yeast adsorbing and removing heavy metal
| Project | Before removing | After removing | Removal efficiency |
| Arsenic (mg/kg) | 2.727 | 1.756 | 35.61% |
Sea cucumber ovum homogenate is not centrifuged into two parts again and handles respectively, but be directly added into after acidolysis in comparative example 1
Fixed yeast gel ball adsorbs heavy metal, and removal efficiency only has 35.61%, in embodiment 3 after the homogenate acidolysis of sea cucumber ovum, centrifugation
It is divided into two parts to handle respectively, i.e., respectively in yeast bacterium mud and fixed yeast gel ball absorption supernatant and substrate precipitating
Heavy metal, removal efficiency is up to 75.31%, than being doubled in comparative example, it is seen that removal methods provided by the invention can be obvious
Improve the removal efficiency of heavy metal.
As can be seen from Figure 2 fixed yeast gel ball is milky, the gel ball that diameter is about 2mm, it is easy into
Separation after row removing.
From figure 3, it can be seen that when the pH of hydrochloric acid solution is between 2.5~3.5, weight after acidolysis marine animal homogenate
The removal efficiency of metal can reach 90% or more, when concentration of hydrochloric acid is too low, and heavy metal removing rate is decreased obviously.
Figure 4, it is seen that removal efficiency obviously increases when adjustment supernatant pH is 8, when possible cause is pH too low,
It is unfavorable for Yeast Growth, is also unfavorable for saccharomycete and absorbs heavy metal.Under conditions of pH is 8, Pichia kudriavezii uses 8%
After sodium chloride preculture, removal efficiency highest.
Sea cucumber intestine homogenate nutriment yield after acidolysis-yeast absorption removing heavy-metal is 89% in embodiment 2, is shown
The rate of recovery (generally below 50%) for being higher than conventional removing heavy-metal method is write, the loss of nutriment is greatly reduced.
Claims (9)
1. a kind of marine animal homogenate removing heavy-metal method, which is characterized in that expand culture, prepare to saccharomycete first
Fixed yeast gel ball, then prepare marine animal homogenate；Acidolysis marine animal homogenate simultaneously carries out solid-liquid separation treatment, most
Afterwards using the heavy metal in precipitating after the removing separation of yeast gel ball, and using in supernatant after free yeast bacterium adsorbing separation
Heavy metal.
2. removing heavy-metal method as described in claim 1, which is characterized in that the saccharomycete includes Pichia kudriavezii, makes
One or more mixing of brewer yeast, Lu Shi yeast, brewer's yeast.
3. removing heavy-metal method as described in claim 1, which is characterized in that above-mentioned saccharomycete is right during expanding culture
Saccharomycete carries out osmotic pressure Stress treatment, and osmotic pressure stress conditions include NaCl Stress or/and carbohydrate stress.
4. removing heavy-metal method as claimed in claim 3, which is characterized in that the carbohydrate stress is sucrose stress.
5. removing heavy-metal method as described in claim 1, which is characterized in that the acid solution in the acid hemolysis process includes salt
The one or more mixing of acid, phosphoric acid, lactic acid.
6. removing heavy-metal method as described in claim 1, which is characterized in that its specific implementation step includes:
1) saccharomycete activation and the preparation of yeast paste
Saccharomycete successively carry out solid activation and liquid activation, take liquid activate after yeast bacteria suspension be added sodium chloride-containing and/
Or in the fluid nutrient medium of sugar, shake culture；After culture, fermentation liquid is centrifuged, is flocculated or film filtration treatment, and
Yeast bacterium mud is obtained after cleaning；
2) preparation of fixed yeast gel ball
Sodium alginate soln is prepared, then yeast bacterium mud and sodium alginate prepared by step 1) are after mixing, using granulation skill
Art prepares fixed yeast gel ball, and gel media is calcium chloride solution, stands cleaning, it is standby to obtain fixed yeast gel ball
With；
3) the acidolysis processing of marine animal homogenate
Marine animal is homogenized after pre-processing according to a conventional method, obtains marine animal homogenate；The acid solution of pH0.5~pH6 is added
The mass volume ratio of stirring, marine animal homogenate and acid solution is 1:1~1:20, after reaction, then stands, is then centrifuged for
Or film filtering, supernatant and bottom precipitation are collected respectively；
4) in marine animal homogenate supernatant heavy metal removing
The yeast bacterium mud that step 1) is obtained is added in the marine animal homogenate supernatant of step 3) preparation, adjusts supernatant
PH is 2~10,10~35 DEG C, stirs 5~240min, removes saccharomycete, the supernatant after obtaining heavy metal removing；
5) in marine animal homogenate bottom precipitation heavy metal removing
Fixed yeast gel ball prepared by step 2) is added in the marine animal homogenate bottom precipitation that step 3) obtains,
It adds water and stirs, adjusting pH is 2~10,10~35 DEG C, 0.2~8h of shake culture, removes fixed yeast gel ball, obtains bottom
It precipitates and carries out nutriment recycling.
7. removing heavy-metal method as claimed in claim 6, which is characterized in that the concentration of sodium chloride is in the step 1)
8wt%.
8. removing heavy-metal method as claimed in claim 6, which is characterized in that acid solution is hydrochloric acid, pH in the step 3)
It is 3.
9. removing heavy-metal method as claimed in claim 6, which is characterized in that carry out removing weight in the step 4) and step 5)
The pH of metal is 8.
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| CN109793149A (en) * | 2019-03-02 | 2019-05-24 | 大连棒棰岛海产股份有限公司 | Sea cucumber intestine extract liquid quelite and its preparation method |
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