CN103156049A - Method for extracting protein and chitin by fermenting shrimp heads and shrimp shells - Google Patents
Method for extracting protein and chitin by fermenting shrimp heads and shrimp shells Download PDFInfo
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- CN103156049A CN103156049A CN2011104074778A CN201110407477A CN103156049A CN 103156049 A CN103156049 A CN 103156049A CN 2011104074778 A CN2011104074778 A CN 2011104074778A CN 201110407477 A CN201110407477 A CN 201110407477A CN 103156049 A CN103156049 A CN 103156049A
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- 241000238557 Decapoda Species 0.000 title claims abstract description 100
- 229920002101 Chitin Polymers 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 35
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 30
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 30
- 238000000855 fermentation Methods 0.000 claims abstract description 56
- 230000004151 fermentation Effects 0.000 claims abstract description 56
- 240000002605 Lactobacillus helveticus Species 0.000 claims abstract description 43
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- 238000011081 inoculation Methods 0.000 claims description 11
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- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 claims description 2
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Abstract
The invention discloses a method for extracting protein and chitin by fermenting shrimp heads and shrimp shells. The method comprises the following steps of: (1) adding a carbon source and a nitrogen source in the shrimp heads and shrimp shells which are crushed or homogenized to be 10-50 meshes, and then uniformly mixing; (2) inoculating lactobacillus helveticus or a mixed bacterial strain; (3) performing heat-insulating fermentation; (4) separating the fermentation broth from residue after the fermentation is concluded; (5) drying the fermentation broth to obtain protein powder; and (6) soaking the fermentation residue obtained in the step (4), and decolouring and fermenting to obtain chitin. With the adoption of the method disclosed by the invention, the protein powder without fishiness and rich in calcium lactate, and chitin can be obtained; and the method is rapid, simple, and capable of reducing production cost, reducing environmental pollution, and adequately utilizing rich nutritional ingredients in the shrimp heads and the shrimp shells.
Description
Technical field
The present invention is specifically related to a kind of method of extracting protein and chitin by fermented shrimp head and shrimp shell.
Background technology
Concerning shrimps, shrimp head and shrimp shell are rich in abundant nutriment, for example the shrimp head of prawn, shrimp shell account for 30% left and right of shrimp body weight, wherein except containing the mineral matters such as chitin, calcium carbonate, also contain the compositions such as 7-15% protein, 2-4% grease, abundant vitamin E, lutein, lecithin, nutritive value is higher.
The at present utilization of prawn head, shrimp shell mainly two sides and: the one, for the production of chitin, the 2nd, utilize autologous tissue's protease or foreign protein enzyme hydrolysis protein wherein, its liquefaction is separated water outlet.But directly produce chitin with shrimp head, shrimp shell, need acid, alkali fermentation raw material with high concentration, not only seriously polluted, and wherein, protein, fat, calcareous etc. all be removed as impurity, can't utilize.The hydrolyzate that method by protease hydrolytic obtains also all exists obvious bitter taste and stench flavor, and product special flavour is poor, can't be as the base-material of other food, can only make feed, in addition, be easy to occur corrupt in hydrolytic process, and add protease and also improved production cost.
The at present domestic existing report that utilizes lactobacillus-fermented shrimp head, shrimp shell to extract active material, one or more mixing of bacterial strain uses therefor such as Lactobacillus plantarum, lactobacillus acidophilus, not yet utilize Lactobacillus helveticus, perhaps comprise Lactobacillus helveticus at the similar report of interior hybrid bacterial strain.Although a small amount of similarly research report is abroad arranged, but be mainly to utilize lactobacillus-fermented can produce the character of a large amount of lactic acid, the pH value of shrimp head, shrimp shell is reduced greatly, reach nature corrosion-resistant purpose, make raw material be able to long-term storage at normal temperatures, be convenient to the ensilage as animal.Be exactly to utilize lactobacillus-fermented that proteolysis liquefaction in shrimp head, shrimp shell is separated in addition, the compositions such as simultaneously a small amount of fat, astaxanthin also dissociate out, the lactic acid of fermenting and producing is also with the calcareous stripping in shell, due to shrimp head, the decalcification of shrimp shell, produce chitin with the fermentation residue that filters out easier.
as Agricultural University Of South China. a kind of method [P] of extracting protein and chitin from shrimp head and shrimp shell. China: CN 101579132A, any one or the three's that utilize lactobacillus acidophilus, streptococcus thermophilus, lactobacillus delbruockii subspecies bulgaricus who reports in 2009.11.18. Mixed Microbes is processed, a kind ofly utilize Pediococcus acidilactici to extract the method [P] of protein and chitin from shrimp head and shrimp shell. China: CN101869170A, 2010.05.18. one kind is utilized Lactobacillus plantarum to extract the method [P] of protein and chitin from shrimp head and shrimp shell. China: CN 101869171A, 2010.05.18.D.J.Evers and D.J.Carroll (D.J.Evers, D.J.Carroll.Ensiling salt-preserved shrimp waste with grass straw and molasses.Animal feed Science Technology.1998, 71:241-249) having studied mixing utilizes being combined of Lactobacillus helveticus and enterococcus faecalis to add molasses and rice straw fermented shrimp head, the shrimp shell is used for making animal feed.people (the Neith Pacheco such as Neith PacHeco, Monica Garnica-Conzalez, Jessica Y.Ramfrez-Hernandez, et al.Effect of temperature on chitin and astaxanthin recoveries f ' rom shrimp waste using lactic acid bacteria, Bioresource, Technology, 2009, 100:2849-2854) studied and be utilized as the Lactobacillus plantarum fermented shrimp shell, the shrimp head reclaims chitin, or utilize the hybrid bacterial strain fermented shrimp shell, the shrimp head reclaims astaxanthin and chitin, and studied the impact of temperature on ferment effect, the people such as N.M.Sachindra (N.M.Sachindra, N.Bhaskar, G..S.Siddegowda.et al.Recovery of carotenoids from ensilaged shrimp waste[J] .Bioresource Technology, 2007,98:1642-1646) studied and utilize Lactobacillus helveticus or hybrid bacterial strain to reclaim the problem of carotenoid from the zymotic fluid of shrimp shell, shrimp head.People (the Luis A.Cira such as Luis A.Cira, Sergio huerta, George M.hall, et al.Pilot scale lactic acid fermentation of shrimp wastes for chitin recovery.Process Biochemistry, 2002,37:1359-1366) studied the optimal conditions of fermentation of lactobacillus ferment shrimp shell, shrimp head, comprise inoculum concentration, add sugar type, sugaring amount, fermentation time etc., they adopt sucrose to make carbon source, inoculum concentration 5%, 30 ℃ of fermentation temperatures, fermenting, after 96 hours, system pH can be reduced to 4.2-4.3.
Summary of the invention
Technical problem to be solved by this invention is in order to overcome exist in the existing method of extracting protein and chitin from aquatic products seriously polluted, the problem that nutrient loss and treatment effeciency are low, and a kind of method of extracting protein and chitin by fermented shrimp head and shrimp shell is provided.Method of the present invention can obtain the stench flavor of nothing and be rich in the protein powder of calcium lactate, and chitin, and method is simple fast, has reduced production cost, and environmental contamination reduction takes full advantage of the nutritional labeling that is rich in shrimp head and shrimp shell.
Therefore, the present invention relates to a kind of method of extracting protein and chitin by fermented shrimp head and shrimp shell, it comprises the following step:
(1) add carbon source, nitrogenous source and mix in pulverizing or homogenate to 10 order～50 purpose shrimp heads and shrimp shell;
(2) inoculation Lactobacillus helveticus or hybrid bacterial strain;
(3) heat-preservation fermentation;
(4) after fermentation ends, zymotic fluid is separated with residue;
(5) zymotic fluid is dry, obtain protein powder;
(6) fermentation residue with step (4) gained soaks and the decolouring processing through diluted acid, obtains chitin;
Wherein, described hybrid bacterial strain is the hybrid bacterial strain of hybrid bacterial strain, Lactobacillus helveticus and streptococcus thermophilus of Lactobacillus helveticus and lactobacillus bulgaricus or the hybrid bacterial strain of Lactobacillus helveticus and lactobacillus bulgaricus and streptococcus thermophilus, and hybrid bacterial strain is cultivated separately respectively and combined inoculation; In the hybrid bacterial strain of Lactobacillus helveticus and lactobacillus bulgaricus, the inoculative proportion of Lactobacillus helveticus and lactobacillus bulgaricus is 1: 2-2: 1 (preferred 1: 1); In the hybrid bacterial strain of Lactobacillus helveticus and streptococcus thermophilus, the inoculative proportion of Lactobacillus helveticus and streptococcus thermophilus is 1: 2-2: 1 (preferred 1: 1); In the hybrid bacterial strain of Lactobacillus helveticus and lactobacillus bulgaricus and streptococcus thermophilus, the inoculative proportion of Lactobacillus helveticus and lactobacillus bulgaricus and streptococcus thermophilus is 2: 1: 1;
In the step (2) inoculation be Lactobacillus helveticus or hybrid bacterial strain the time, the shrimp head in step (1) and shrimp shell are shrimp head fresh or cold storage and shrimp shell, when shrimp head and shrimp shell were the shrimp head of cold storage and shrimp shell, need thawed shrimp head and the shrimp shell of cold storage.
Wherein, step (5) and (6) can be carried out simultaneously, perhaps carry out according to the order of step (5), (6) or step (6), (5).
In the present invention, in step (1), the described method that shrimp head and the shrimp shell of cold storage are thawed can be the defreezing method of various routines, as naturally thaw, hot-air thaws, flowing water thaws, microwave thawing or electrical heating are thawed etc.
In step (1), the addition of carbon source can be the 5%-30% of shrimp head and shrimp shell gross mass, preferred 12%-20%.These carbon sources can comprise one or more in glucose, sucrose, maltose, dextrin, starch, starch syrup, HFCS, lactose, whey powder and molasses.The addition of nitrogenous source can be 0.1%～10% of shrimp head and shrimp shell gross mass, preferred 0.5-2%.These nitrogenous sources can comprise one or more in ammonium sulfate, ammonium nitrate, ammonium chloride, urea, peptone, dusty yeast and beancake powder etc.
In step (2), described Lactobacillus helveticus or hybrid bacterial strain are those skilled in the art's fermenting and producing varieties of food items probio commonly used, and sale is arranged on the market.The inoculation of Lactobacillus helveticus or mixed bacteria adopts those skilled in the art's routine operation to carry out, and the inoculum concentration of Lactobacillus helveticus can be 1%～20% of shrimp head and shrimp shell gross mass, preferred 5%-15%.
In the heat-preservation fermentation step of step (3), for improving ferment effect, can also add appropriate water in shrimp head, shrimp shell, generally add the water of 20%-200% of shrimp head and shrimp chitin amount better.For shortening the induction period of fermentation, can regulate the pH of shrimp head and shrimp shell with the acid that allows in food or feed to use, reach between 6.0-6.8.
In step (3), the temperature of described heat-preservation fermentation can be selected 20-50 ℃, preferred 35-45 ℃; More than fermentation time can be 18h, be no more than 120h, preferred 48-72h.
In step (4), after fermentation ends, can (because step (1) Prawn head and shrimp shell are 10 orders～50 orders, therefore filtering needs with the higher screen pack of order number by centrifugal or filter method, residue obtained for removing most of calcareous shrimp shell, produce the raw material of chitin as next step.Zymotic fluid is separated with fermentation residue.For removing the tart flavour of zymotic fluid, the alkali that can adopt those skilled in the art to commonly use is as adopting sodium carbonate or sodium acid carbonate neutralise broth.
In step (5), the drying means that the drying of zymotic fluid can adopt any one those skilled in the art to commonly use, one or more in as natural drying in adopting, oven dry and spray-drying obtain powdered substance after drying, be the protein of gained of the present invention.
In step (6), adopting diluted acid to remove relict mineral matter and decolouring fermentation chitin extraction, is the practices well that those skilled in the art obtain chitin.The present invention during chitin extraction, selects diluted acid soaking fermentation residue from fermentation residue, can select various diluted acids, comprises various inorganic acids or organic acid, and as watery hydrochloric acid, the concentration of diluted acid is higher, and soak time is longer, and the effect of demineralization is just better.Therefore diluted acid can be selected 0.01mol/L-1mol/L in the concentration of hydrogen ion degree (as watery hydrochloric acid), preferred 0.2-0.5mol/L; Soak time can be selected 0.5h-6h, preferred 2-4h.Decolouring after diluted acid soaks is processed, adopt the decolouring fermentation operation that in prior art, chitin extraction is commonly used all can realize the present invention, as adopting conventional hydrogen peroxide decolouring, potassium permanganate decolouring or sunlight exposure etc. can obtain required chitin at last from fermentation residue.
Except specified otherwise, the raw material that the present invention relates to and reagent is commercially available getting all.
Positive progressive effect of the present invention is:
1, the present invention's high acid environment of utilizing the fermentation of Lactobacillus helveticus or hybrid bacterial strain to cause can prevent that shrimp head, shrimp shell from corruption occuring in the self-dissolving process, thereby the self-dissolving time can be extended greatly, the a small amount of protease that produces in the prolongation of self-dissolving time and Lactobacillus helveticus sweat, can guarantee abundant hydrolysis and the high usage of protein, need not to add protease, peracid is also very favourable for follow-up fermentation and the preservation of product in addition again;
2, the present invention's lactic acid of utilizing fermentation to produce can remove the mineral matter major parts such as calcium salt in shrimp head, shrimp shell, with the shrimp head after demineralization, that the shrimp shell is produced chitin is easier, and can reduce the consumption of hydrochloric acid, reduces costs and environmental contamination reduction;
3, the present invention utilizes Lactobacillus helveticus or hybrid bacterial strain fermentation can eliminate the stench flavor of zymotic fluid, can improve the palatability of product, and zymotic fluid is rich in the mineral matters such as calcareous;
4, the present invention adopts Lactobacillus helveticus or hybrid bacterial strain zymotechnique, not only method of operating is simple, low cost of raw materials, and can be simultaneously obtain protein and chitin from shrimp head and shrimp shell makes that in shrimp head and shrimp shell, contained composition is fully utilized.
5, the present invention's shrimp head used, shrimp shell be all through pulverizing or homogenate to 10 order-50 order, and be thinner, not only can fully enlarge the effect contact area of the bacterial strains such as Lactobacillus helveticus, also can reduce fermentation time, improves treatment effect.
6, the present invention's rear residue that ferments is further processed with diluted acid, can guarantee that the impurity content of solid product remains in the scope of requirement.Diluted acid after processing simultaneously is further with the alkali neutralization, and assorted through reverse-osmosis treated desalination etc., and recyclable salt is as feed addictive use, and sewage also reaches discharge standard.
The specific embodiment
The below further illustrates the present invention with embodiment, but the present invention is not limited.
Embodiment 1
The present embodiment utilizes the Lactobacillus helveticus fermentation to extract protein and chitin from shrimp head and shrimp shell, and its concrete steps are as follows:
(1) get the fresh shrimp head of 50g and shrimp shell, shrimp head and shrimp shell meal are broken to 10 orders left and right, add 25mL water (shrimp head and shrimp shell gross mass 50%), survey joint pH to 6.4;
(2) add again 7.5g glucose (shrimp head and shrimp shell gross mass 15%), the dusty yeast of 0.5g (shrimp head and shrimp shell gross mass 1%);
(3) inoculation Lactobacillus helveticus, inoculum concentration is 10% of shrimp head and shrimp shell gross mass;
(4) in 37 ℃ of heat-preservation fermentations 3 days;
(5) after fermentation ends, centrifugation zymotic fluid and fermentation residue; Zymotic fluid neutralizes through sodium carbonate, and is concentrated, obtains powdered substance 7.4g after spray-drying, and this powdered substance is the present embodiment gained protein;
(6) fermentation residue after 0.5mol/L watery hydrochloric acid soaks 5h, is striven neutrality with the clear water rinsing, is put in the decolouring of being exposed to the sun under daylight, obtains chitin.
Embodiment 2
The present embodiment utilizes the Lactobacillus helveticus fermentation to extract protein and chitin from shrimp head and shrimp shell, and its concrete steps are as follows:
(1) get the fresh shrimp head of 100g and shrimp shell, shrimp head and shrimp shell meal are broken to about 10 orders, survey joint pH to 6.3;
(2) add again 20g glucose (shrimp head and shrimp shell gross mass 20%), the dusty yeast of 2g (shrimp head and shrimp shell gross mass 2%);
(3) inoculation Lactobacillus helveticus, inoculum concentration is 5% of shrimp head and shrimp shell gross mass;
(4) in 37 ℃ of heat-preservation fermentations 3 days;
(5) after fermentation ends, centrifugation zymotic fluid and fermentation residue; Zymotic fluid neutralizes through sodium carbonate, and is concentrated, obtains powdered substance 14.9g after spray-drying, and this powdered substance is the present embodiment gained protein;
(6) fermentation residue soaks 6h through 0.4mol/L watery hydrochloric acid, then floats Shen to neutral with clear water, is put in the decolouring of being exposed to the sun under daylight, obtains chitin.
Embodiment 3
The present embodiment utilizes the hybrid bacterial strain fermentation to produce protein and chitin from shrimp head and shrimp shell, and its concrete steps are as follows:
(1) get shrimp head and the shrimp shell of 50g drying, shrimp head and shrimp shell meal are broken to 10 orders left and right, add 25mL water (shrimp head and shrimp shell gross mass 50%), survey joint pH to 6.4;
(2) add again 7.5g glucose (shrimp head and shrimp shell suffer from quality 15%), the dusty yeast of 1g (shrimp head and shrimp shell gross mass 2%);
(3) inoculation Lactobacillus helveticus: lactobacillus bulgaricus=1: 1, inoculum concentration are 10% of shrimp head and shrimp shell gross mass;
(4) in 42 ℃ of heat-preservation fermentations 3 days;
(5) after fermentation ends, centrifugation zymotic fluid and fermentation residue; Zymotic fluid is produced astaxanthin after ethanol extraction, the water after extraction neutralizes through sodium carbonate, and is concentrated, obtains powdered substance 7.4g after spray-drying, and this powdered substance is the present embodiment gained protein;
(6) fermentation residue after 0.5mol/L watery hydrochloric acid soaks 5h, is striven neutrality with the clear water rinsing, is put in the decolouring of being exposed to the sun under daylight, obtains chitin.
Embodiment 4
The present embodiment utilizes the hybrid bacterial strain fermentation to produce protein and chitin from shrimp head and shrimp shell, and its concrete steps are as follows:
(1) get shrimp head and the shrimp shell of 100g drying, with shrimp head and shrimp shell meal broken 10 about orders of striving, survey joint pH to 6.3;
(2) add again 20g glucose (shrimp head and shrimp shell gross mass 20%), the dusty yeast of 1g (shrimp head and shrimp shell gross mass 1%);
(3) inoculation Lactobacillus helveticus: lactobacillus bulgaricus: thermophilic hammer=2: 1: 1, inoculum concentration are 10% of shrimp head and shrimp shell gross mass;
(4) in 42 ℃ of heat-preservation fermentations 3 days;
(5) after fermentation ends, centrifugation zymotic fluid and fermentation residue; Zymotic fluid is produced astaxanthin after ethanol extraction, the water after extraction neutralizes through sodium carbonate, and is concentrated, obtains powdered substance 14.9g after spray-drying, and this powdered substance is the present embodiment gained protein;
(6) fermentation residue soaks 6h through 0.4mol/L watery hydrochloric acid, then floats Shen to neutral with clear water, is put in the decolouring of being exposed to the sun under daylight, obtains chitin.
Claims (10)
1. method of extracting protein and chitin by fermented shrimp head and shrimp shell is characterized in that comprising the following step:
(1) add carbon source, nitrogenous source and mix in pulverizing or homogenate to 10 order～50 purpose shrimp heads and shrimp shell;
(2) inoculation Lactobacillus helveticus or hybrid bacterial strain;
(3) heat-preservation fermentation;
(4) after fermentation ends, zymotic fluid is separated with residue;
(5) zymotic fluid is dry, obtain protein powder;
(6) fermentation residue with step (4) gained soaks and the decolouring processing through diluted acid, obtains chitin;
Wherein, described hybrid bacterial strain is the hybrid bacterial strain of hybrid bacterial strain, Lactobacillus helveticus and streptococcus thermophilus of Lactobacillus helveticus and lactobacillus bulgaricus or the hybrid bacterial strain of Lactobacillus helveticus and lactobacillus bulgaricus and streptococcus thermophilus, and hybrid bacterial strain is cultivated separately respectively and combined inoculation; In the hybrid bacterial strain of Lactobacillus helveticus and lactobacillus bulgaricus, the inoculative proportion of Lactobacillus helveticus and lactobacillus bulgaricus is 1: 2-2: 1; In the hybrid bacterial strain of Lactobacillus helveticus and streptococcus thermophilus, the inoculative proportion of Lactobacillus helveticus and streptococcus thermophilus is 1: 2-2: 1; In the hybrid bacterial strain of Lactobacillus helveticus and lactobacillus bulgaricus and streptococcus thermophilus, the inoculative proportion of Lactobacillus helveticus and lactobacillus bulgaricus and streptococcus thermophilus is 2: 1: 1;
In the step (2) inoculation be Lactobacillus helveticus or hybrid bacterial strain the time, the shrimp head in step (1) and shrimp shell are shrimp head fresh or cold storage and shrimp shell, when shrimp head and shrimp shell were the shrimp head of cold storage and shrimp shell, need thawed shrimp head and the shrimp shell of cold storage;
Wherein, carry out simultaneously step (5) and (6), perhaps carries out according to the order of step (5), (6) or step (6), (5).
2. the method for claim 1 is characterized in that: in step (1), the described method that shrimp head and the shrimp shell of cold storage are thawed is that nature thaws, hot-air thaws, flowing water thaws, microwave thawing or electrical heating are thawed.
3. the method for claim 1, it is characterized in that: in step (1), the addition of carbon source is the 5%-30% of shrimp head and shrimp shell gross mass; Described carbon source comprises one or more in glucose, sucrose, maltose, dextrin, starch, starch syrup, HFCS, lactose, whey powder and molasses.
4. method as claimed in claim 3, it is characterized in that: in step (1), the addition of carbon source is the 12%-20% of shrimp head and shrimp shell gross mass.
5. the method for claim 1, it is characterized in that: in step (1), the addition of nitrogenous source is 0.1%～10% of shrimp head and shrimp shell gross mass; These nitrogenous sources comprise one or more in ammonium sulfate, ammonium nitrate, ammonium chloride, urea, peptone, dusty yeast and beancake powder.
6. the method for claim 1, it is characterized in that: in step (2), the inoculum concentration of described Lactobacillus helveticus is 1%～20% of shrimp head and shrimp shell gross mass.
7. method as claimed in claim 6, it is characterized in that: in step (2), the inoculum concentration of described Lactobacillus helveticus is shrimp head and shrimp shell gross mass 5%-15%.
8. the method for claim 1 is characterized in that: in step (3), also add the water of the 20%-200% of shrimp head and shrimp chitin amount in shrimp head, the shrimp shell; Regulate the pH of shrimp head and shrimp shell with the acid that allows in food or feed to use, reach between 6.0-6.8.
9. the method for claim 1, it is characterized in that: in step (3), the temperature of described heat-preservation fermentation is 20-50 ℃; Fermentation time is 18h-120h.
10. the method for claim 1, is characterized in that: in step (4), after fermentation ends, adopt sodium carbonate or sodium acid carbonate neutralise broth; In step (6), described diluted acid is take the concentration of hydrogen ion degree as 0.01mol/L-1mol/L; Soak time is selected 0.5h-6h; Decolouring after diluted acid soaks is processed, and adopts conventional hydrogen peroxide decolouring, potassium permanganate decolouring or sunlight exposure method.
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