CN104522798B - A kind of fermentation krill beverage and preparation method thereof - Google Patents
A kind of fermentation krill beverage and preparation method thereof Download PDFInfo
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Abstract
The object of this invention is to provide a kind of fermentation krill beverage and preparation method thereof, is prepare as base fluid with krill fermented liquid prepared by morel and fermentation of bacillus subtilis krill.The present invention to be fermented krill by morel and subtilis, do not need during the fermentation to add soda acid or commercial enzyme, the proteolytic enzyme utilizing morel and subtilis to produce during the fermentation, chitinase isoreactivity enzyme can make krill proteolytic degradation be more easily absorb and solvable polypeptide and amino acid, and other nutritive substances are as effective in astaxanthin and various trace element have also been obtained to be dissolved and release.Morel also can produce tool activated biological polyoses class material during the fermentation, enriches the nutrition of fermented drink.The environmental protection of whole drink fermented process is pollution-free, and containing the functional component such as polypeptide, astaxanthin, polysaccharide useful to human body, often drinks and can realize comprehensive extra-nutrition, improves the effects such as immunizing power, antifatigue, coordinating intestines and stomach.
Description
Technical field
The invention belongs to field of foodstuff beverage, be specifically related to a kind of fermentation krill beverage and preparation method thereof.
Background technology
Along with the development of society, growth in the living standard, human consumer has been not content with the single common beverages of nutrition, to more and more urgent through biotechnology processing, the nutritious demand having the nourishing drink of health-preserving function concurrently.
Krill is one of single biology that on the earth, stock number is maximum, is considered to maximum marine protein storehouse.Krill is aboundresources not only, and has very high nutritive value.According to surveying and determination, krill fresh shrimp protein matter content is 11.9-15.4%, and containing the necessary whole amino acid of human body, and the ratio of indispensable amino acid and non-essential amino acid is suitable for, and is more conducive to absorption and the utilization of human body.Simultaneously also be rich in biologically active substance and the medicinal raw materials such as lipid, astaxanthin, chitin in krill body, there is the health-care effecies such as anti-oxidant, anti-ageing, antifatigue, radioprotective and raising immunizing power.The main edible way of present stage krill is directly edible and edible as peeled shrimp after shelling, and edible conditions is restricted and much nutrition is difficult to absorbed intact.If krill to be developed to nutritious, that instant, assimilated efficiency are high drink product is inherently subject to human consumer favor.
Summary of the invention
The object of this invention is to provide a kind of fermentation krill beverage and preparation method thereof, i.e. a kind of nutritious, protein drinks with health-care effect, make up that market existing beverage category is single, the situation of nutrient deficiency.
First the present invention provides a kind of krill fermented liquid, utilizes morel (Morehellaesculenta) and subtilis (Bacillussubtilis) krill that ferments to prepare;
Above-mentioned krill fermented liquid, its a kind of concrete preparation process is as follows:
1) choose krill and carry out degreasing, make krill spent meal;
2) use step 1) in degreasing krill meal to be mixed with mass percent concentration be that the krill meal aqueous solution of 1-3% is as morel activation medium;
3) morel is linked in morel activation medium and carries out activation and obtain morel activation culture liquid,
4) by step 3) in the morel activation culture liquid that activated transfer in toadstool ferment substratum and ferment, obtain morel krill fermented liquid;
The krill meal aqueous solution of to be mass percent concentration the be 3-10% of the toadstool ferment substratum in above-mentioned steps (4); Wherein fermentation condition is 20-30 DEG C, 150-200rpm oscillation and fermentation 5-10 days;
5) the degreasing krill meal in step (1) is used to be mixed with subtilis activation medium,
6) subtilis is accessed in subtilis activation medium activate, make bacillus subtilis bacteria culture fluid;
The wherein krill meal aqueous solution of subtilis activation medium to be mass percent concentration be 3-5%;
7) by step 6) in bacillus subtilis bacteria culture fluid to transfer step 4) in the morel krill fermented liquid that ferments ferment, make fermented liquid;
Fermentation condition is 30-37 DEG C, under 150-200rpm concussion cultivate 3-5 days;
8) by step 7) in cleer and peaceful residue on broken, the extracting and developing of cultured fermented liquid, the supernatant liquor obtained is krill fermented liquid.
Described fragmentation utilizes refiner to smash mycelium fermented liquid, and described extraction is extracted at 50 DEG C-80 DEG C water-bath 5-8 hour.
Above-mentioned krill fermented liquid is used as the base fluid of fermented drink;
The present invention also provides a kind of krill beverage prepared as base fluid with above-mentioned krill fermented liquid; Be that fermented drink base fluid is carried out sterilising treatment, add jujube juice, beta-cyclodextrin, sucrose, perfume ingredient be mixed with krill beverage, in beverage, be rich in krill albumen, polypeptide, amino acid and Morchella esculenta (L.) Pers polysaccharide.
The present invention to be fermented krill by morel and subtilis, do not need during the fermentation to add soda acid or commercial enzyme, the proteolytic enzyme utilizing morel and subtilis to produce during the fermentation, chitinase isoreactivity enzyme can make krill proteolytic degradation be more easily absorb and solvable polypeptide and amino acid, and other nutritive substances are as effective in astaxanthin and various trace element have also been obtained to be dissolved and release.Morel also can produce tool activated biological polyoses class material during the fermentation, enriches the nutrition of fermented drink.The environmental protection of whole drink fermented process is pollution-free, and containing the functional component such as polypeptide, astaxanthin, polysaccharide useful to human body, often drinks and can realize comprehensive extra-nutrition, improves the effects such as immunizing power, antifatigue, coordinating intestines and stomach.
Embodiment
Morel is a kind of famous and precious medicine-food two-purpose fungi, nutritious and have antifatigue, improve the health-care effect such as immunizing power, useful stomach; Subtilis can produce the abundant enzyme such as proteolytic enzyme, lipase in liquid culture process; Utilize morel and subtilis to ferment to krill, by the protein delivery that is combined with the position such as shrimp shell in krill out, and the polypeptide and amino acid that are more conducive to absorbing can be decomposed into further.Morel can produce bioactive polysaccharide in process of growth, improves the nutritive value of krill fermented liquid further.Meanwhile, morel and subtilis can also reduce the distinctive fishy smell of krill during the fermentation, make krill fermented liquid local flavor softer, therefore utilize morel and fermentation of bacillus subtilis krill producing drink to have good application and development prospect.
The morel bacterial classification that the specific embodiment of the invention uses and Bacillus subtilis strain can be purchased from China General Microbiological culture presevation administrative center, also can select the bacterial classification in other sources conventional in field of food.
Embodiment 1
(1) preparation of degreasing shrimp med: by purchase come krill first clean up with tap water, after oven dry with pulverizer by broken for dry shrimp med.Take shrimp med 25g, first add 4-5mL water, then add 200mL chloroform-methanol and to be placed on Milko-Tester 65-70 DEG C of heating in water bath backflow, timing from micro-boiling, extract 6.5h, filter, remove residual solvent, obtain degreasing shrimp med after drying and (contrast non-degreasing shrimp med, degreasing shrimp med look light, coarse, fishy smell is lower, without greasy feeling).For the degreasing of shrimp, also can adopt existing prawn degreasing operation to carry out.
(2) the dull and stereotyped seed activation of morel: the bacterial classification cutting inclined-plane or dull and stereotyped upper preservation is about 0.25cm
2, be connected on new potato dextrose agar flat board, cultivate 7 days, treat that Morciiella Esculeuta Mycelia is paved with flat board for 25 DEG C.
(3) morel activation: smashed (adding about 100mL sterilized water) by dull and stereotyped for the morel grow seed with refiner, accesses in the activation medium containing 1% krill meal by 10% inoculum size, 25 DEG C of shaking culture 5 days.
(4) toadstool ferment: the morel nutrient solution activated is forwarded in the fermention medium containing 7% krill meal by 10% inoculum size, 25 DEG C of shaking culture 5 days.
(5) the dull and stereotyped seed activation of subtilis: get a transfering loop thalline from inclined-plane or the dull and stereotyped bacterial classification preserved, rule on LB plate culture medium, cultivate 3 days, treat flat board grows bacterium colony for 30 DEG C.
(6) subtilis activation: get single colony inoculation in the activation medium containing 3% krill from the flat board grow, 30 DEG C of shaking culture 3 days.
(7) fermentation of bacillus subtilis: the bacillus subtilis bacteria culture fluid activated is forwarded in fermented morel liquid by 10% inoculum size, 30 DEG C of shaking culture 3 days.
(8) broth extraction: utilize refiner to be smashed by fermented liquid, in 60 DEG C of shaking bath 7h.
(9) separation of fermentative broth: by the krill fermented liquid that extracted in the centrifugal 15min of 8000rpm, get supernatant.
(10) Beverage Service: get 1Kg fermentation clear liquid, 60g sucrose, 50 jujube juices, 10g beta-cyclodextrin, 0.3g food flavour, mix, with 100 order filter-cloth filterings, the filling sterilization of filtrate can obtain the fermentation krill beverage of red jujube taste, wherein containing protein 10.6g/L, polypeptide 8.2g/L, polysaccharide 500mg/L.
Embodiment 2
The preparation of degreasing shrimp med and dull and stereotyped seed activation are with embodiment 1, with refiner, dull and stereotyped for the morel grown seed is smashed (adding about 100mL sterilized water), by 10% inoculum size access containing 0.5% krill meal, in the seed culture medium of 1% oatmeal, 25 DEG C of shaking culture 6 days.The morel seed culture medium activated is forwarded in the fermention medium containing 10% krill meal by 10% inoculum size, 23 DEG C of shaking culture 8 days.
By subtilis access seed culture medium 37 DEG C of activation culture 1 day containing 5% krill, the bacillus subtilis bacteria culture fluid after activation is inoculated in fermented morel liquid by 5%, 30 DEG C of fermentations 5 days.
To ferment complete fermented liquid being smashed, in 70 DEG C of water-bath mechanical shaking extraction 6h.
The morel krill fermented liquid extracted is filtered, get 1Kg filtrate, 70g jujube juice, 65g sucrose, 10g beta-cyclodextrin, 0.5g food flavour, mix, mix, with 100 order filter-cloth filterings, the filling sterilization of filtrate can obtain the fermentation krill beverage of red jujube taste, wherein containing protein 15.8g/L, polypeptide 10.5g/L, polysaccharide 600mg/L.
Embodiment 3
The content of nutritive substance before and after fermentation, to contain 10% krill fermentation liquor beverage.
Soluble protein, polypeptide and polysaccharide content change before and after table 1 krill drink fermented
| Detection | Blank (g/L) | Sample (g/L) | Content (g/L) | Content |
| Soluble protein | 6.5 | 15.8 | 9.3 | 243.1％ |
| Polypeptide | 3.9 | 10.5 | 6.6 | 269.2％ |
| Polysaccharide | 0.2 | 0.6 | 0.4 | 300.0％ |
As can be seen from Table 1, through morel and subtilis double-bacterium ferment, the content of the soluble protein in krill beverage, polypeptide and polysaccharide all improves a lot.The raising of soluble protein adds the nutrient protein value of krill beverage, the raising of polypeptide enables the nutrition of krill beverage be absorbed and utilized by the body quickly, has antifatigue, improves the raising of polysaccharide of immunity effect of krill beverage is got a promotion.
Free aminoacid content change before and after table 2 krill drink fermented
| Title | Abbreviation | Blank (mg/L) | Sample (mg/L) | Amino acid change (mg/L) | Amino acid changes |
| Threonine | Thr | 13.9 | 1.2 | -12.8 | 8.4％ |
| α-amino-isovaleric acid | Val | 16.9 | 105.0 | 88.1 | 620.3％ |
| Methionine(Met) | Met | 1.5 | 137.4 | 135.9 | 9039.5％ |
| Isoleucine | Ile | 13.4 | 85.1 | 71.8 | 637.2％ |
| Leucine | Leu | 1.9 | 171.8 | 169.9 | 9104.8％ |
| Phenylalanine | Phe | 45.9 | 105.2 | 59.3 | 229.1％ |
| Tryptophane | Trp | 0.6 | 53.9 | 53.3 | 9794.5％ |
| Methionin | Lys | 38.1 | 156.0 | 117.9 | 409.4％ |
| Aspartic acid | Asp | 6.9 | 0.1 | -6.8 | 1.1％ |
| Serine | Ser | 9.9 | 0.8 | -9.1 | 8.2％ |
| L-asparagine | Asn | 8.6 | 9.2 | 0.5 | 106.1％ |
| L-glutamic acid | Glu | 6.2 | 4.5 | -1.7 | 72.8％ |
| Proline(Pro) | Pro | 58.9 | 0.5 | -58.4 | 0.9％ |
| Glycine | Gly | 53.5 | 1.6 | -51.9 | 3.0％ |
| L-Ala | Ala | 23.8 | 2.5 | -21.3 | 10.6％ |
| Tyrosine | Tyr | 14.2 | 135.9 | 121.7 | 956.7％ |
| Histidine | His | 1.8 | 88.7 | 86.9 | 4876.6％ |
| Arginine | Arg | 91.6 | 2.4 | -89.3 | 2.6％ |
| Phosphoserine | Pser | 7.6 | 9.1 | 1.5 | 120.1％ |
| Taurine | Tau | 36.4 | 37.9 | 1.5 | 104.0％ |
| α-aminoadipic acid | α-Aaa | 25.9 | 0.8 | -25.1 | 3.0％ |
| Citrulline | Cit | 1.7 | 0.1 | -1.7 | 3.5％ |
| Beta-alanine | β-Ala | 55.7 | 32.7 | -23.0 | 58.7％ |
| Ornithine | Orn | 5.7 | 14.2 | 8.5 | 248.7％ |
| Indispensable amino acid | EAA | 132.2 | 815.5 | 683.3 | 616.8％ |
| Non-essential amino acid | NEAA | 275.5 | 246.2 | -29.3 | 89.4％ |
| Non-protein amino acid | NPAA | 133.1 | 94.7 | -38.3 | 71.2％ |
| Total amino acid content | TAA | 540.7 | 1156.5 | 615.7 | 213.9％ |
Be increased to 1156.5mg/L by the total free aminoacids total amount in the visible sample by fermentation of table 2 by 540.7mg/L, increase 113.9%.α-amino-isovaleric acid, methionine(Met), Isoleucine, leucine, phenylalanine, tryptophane and Methionin wherein in human body eight kinds of indispensable amino acids have and significantly improve, TEAA improves 516.8%, illustrate and utilize morel and subtilis double-bacterium ferment effectively can improve essential amino acid content in krill beverage, increase the nutritive value of krill beverage.
Can find out compared with Other Drinks preparation technology, the present invention utilizes morel and subtilis double-bacterium ferment krill beverage, utilizes the growth metabolism of thalline to improve the nutritive value of krill beverage.Beverage preparation does not need to use proteolytic enzyme, technique greatly reduces the cost using expensive business enzyme to bring, uses the fermenting process of precious fungi to also improve nutrition and the class of drink product.
Claims (10)
1. a krill fermented liquid, is characterized in that, the preparation method of described fermented liquid is as follows:
1) choose krill and carry out degreasing, make krill spent meal;
2) use step 1) in degreasing krill meal to be mixed with mass percent concentration be that the krill meal aqueous solution of 1-3% is as morel activation medium;
3) morel is linked in morel activation medium and carries out activation and obtain morel activation culture liquid,
4) by step 3) in the morel activation culture liquid that activated transfer in toadstool ferment substratum and ferment, obtain morel krill fermented liquid;
5) the degreasing krill meal in step (1) is used to be mixed with subtilis activation medium,
6) subtilis is accessed in subtilis activation medium activate, make bacillus subtilis bacteria culture fluid;
7) by step 6) in bacillus subtilis bacteria culture fluid to transfer step 4) in the morel krill fermented liquid that ferments ferment, make fermented liquid;
8) by step 7) in cleer and peaceful residue on broken, the extracting and developing of cultured fermented liquid, the supernatant liquor obtained is krill fermented liquid.
2. krill fermented liquid as claimed in claim 1, is characterized in that, described step 4) in the krill meal aqueous solution of toadstool ferment substratum to be mass percent concentration be 3-10%.
3. krill fermented liquid as claimed in claim 1, is characterized in that, described step 4) in fermentation condition be 20-30 DEG C, 150-200rpm oscillation and fermentation 5-10 days.
4. krill fermented liquid as claimed in claim 1, is characterized in that, described step 5) in the krill meal aqueous solution of subtilis activation medium to be mass percent concentration be 3-5%.
5. krill fermented liquid as claimed in claim 1, is characterized in that, described step 6) in fermentation condition be 30-37 DEG C, under 150-200rpm concussion cultivate 3-5 days.
6. krill fermented liquid as claimed in claim 1, is characterized in that, described step 8) described in fragmentation be utilize refiner to smash mycelium fermented liquid.
7. krill fermented liquid as claimed in claim 1, is characterized in that, described step 8) described in extraction be extract at 50 DEG C-80 DEG C water-bath 5-8 hour.
8. krill fermented liquid according to claim 1 is as the application of fermented drink base fluid.
9. a krill beverage, is characterized in that, described krill beverage krill fermented liquid according to claim 1 is prepared as fermented drink base fluid.
10. krill beverage as claimed in claim 9, it is characterized in that, described krill beverage is that fermented drink base fluid is carried out sterilising treatment, adds jujube juice, beta-cyclodextrin, sucrose, perfume ingredient be mixed with.
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| CN106561831A (en) * | 2016-10-20 | 2017-04-19 | 华南师范大学 | Shrimp-Chinese caterpillar fungus fermented live bacteria beverage and preparation method thereof |
| CN107841518A (en) * | 2017-12-06 | 2018-03-27 | 华南协同创新研究院 | A kind of krill anti-oxidation peptide and preparation method and application |
| CN109971572B (en) * | 2018-11-28 | 2020-08-14 | 中国海洋大学 | Preparation method of degreased antarctic krill fermented milk beer |
| CN111149963A (en) * | 2020-01-21 | 2020-05-15 | 中国海洋大学 | Health-care krill oil lactobacillus beverage and preparation method thereof |
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| CN101962666B (en) * | 2010-09-06 | 2012-12-12 | 湖北工业大学 | Method for preparing chitin, L-calcium lactate and compound amino acid or peptide from carapace waste |
| CN103224973B (en) * | 2013-04-03 | 2014-09-10 | 中国海洋大学 | Method of fementing shrimp heads to prepare active substances, chitin and organic acidity calcium |
| CN104357361B (en) * | 2014-11-14 | 2015-06-03 | 青岛农业大学 | Bacillus subtilis capable of producing chitinase and application thereof |
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