CN103960699B - A kind of krill peptide replenishers containing calcium iron zinc - Google Patents

Abstract

The object of the present invention is to provide a kind of Antarctic krill peptide supplement containing calcium, iron and zinc. The Antarctic krill peptide supplement containing calcium, iron and zinc is to combine calcium, iron and zinc metal elements into the Antarctic krill powder degreased, Krill peptide powder made by protease degradation and defluorination with CaCl ₂ . The raw materials used in the present invention are rich and easy to obtain, and have high protein content; the preparation method has the advantages of simple process steps, less investment in equipment, no environmental pollution, high product yield, etc., and is suitable for industrial production; the prepared krill peptide-calcium iron zinc supplement The agent has the following characteristics: the protein content is higher than 90%, and the product has high purity; the fluorine content is lower than 10ppm, and the product safety is high; the molecular weight is mainly distributed in 2000-200Da, and the water solubility is good; Can supplement three kinds of nutrients and essential amino acids at the same time.

Description

An Antarctic Krill Peptide Supplement Containing Calcium Iron Zinc

technical field

The invention belongs to the technical field of functional food, and in particular relates to an Antarctic krill peptide supplement containing calcium, iron and zinc, a preparation method and application thereof.

Background technique

Calcium, iron and zinc deficiency is a series of related diseases caused by the lack of calcium, iron and zinc. It has become an increasingly serious social health problem worldwide, especially for the elderly, children and pregnant women. special groups such as women. Zinc deficiency is mainly manifested as stunted growth and development, suppressed immune system function, brain damage in perinatal infants, anorexia, skin infections, and decreased reproductive capacity. Iron deficiency is mainly manifested as weakened bactericidal ability of phagocytes and decreased immunity of the body. Calcium deficiency is mainly manifested in conditions such as osteoporosis, high blood pressure, colon cancer, obesity and kidney stones. According to the 2004 survey on nutrition and health status of Chinese residents, the average prevalence rate of anemia in residents is 15.2%, and the prevalence rates of anemia in infants under 2 years old, elderly people over 60 years old and women of childbearing age are 24.2%, 21.5% and 20.6% respectively; The calcium intake was only 41% of the recommended intake; the growth retardation rate and underweight rate of children under 5 years old were 17.3% and 9.3% respectively.

The relevant enhancer products currently on the market are mainly inorganic metal salts and organic acid-metal salts. Among them, the absorption rate of calcium, iron and zinc in inorganic metal salts and organic acid-metal salts is less than 40%. Research by medical experts has shown that the absorption of the three elements of calcium, iron and zinc in the body can coordinate with each other and promote the absorption of each other. The scientific combination of the three elements can effectively improve the lack of calcium, iron and zinc in the body and meet the body's demand for calcium, iron and zinc. Clinical studies have shown that in the absence of three elements at the same time, supplementing one of them alone cannot achieve improvement. Calcium-iron-zinc compound preparations can increase children's appetite, improve rickets, increase hemoglobin value and promote children's intelligence and healthy growth and development. Therefore, finding a safe and effective nutritional fortifier that can supplement calcium, iron and zinc at the same time is a current research hotspot.

Contents of the invention

The purpose of the present invention is to provide an Antarctic krill peptide supplement containing calcium, iron and zinc, its preparation method and application, so as to make up for the deficiencies in the prior art.

The Antarctic krill peptide supplement containing calcium, iron and zinc of the present invention is prepared by the following method:

1) After the Antarctic krill powder is treated with ethanol, it is dried and pulverized to obtain the defatted krill powder;

2) Add distilled water to defatted krill powder, the mass ratio of solid to liquid is 1:8-12, adjust the pH value of the solution to 7.0-8.0, then add protease to the solution for hydrolysis, heat to terminate the enzymatic reaction after hydrolysis, centrifuge Take supernatant A;

3) Adjust the pH value of the supernatant A to 8.5-9.5, add CaCl ₂ to the supernatant A for defluorination, mix well and let stand, centrifuge to take the supernatant B, and adjust the pH of the supernatant B to 7.0 ~7.5, the supernatant B was desalted by nanofiltration membrane, and spray-dried to obtain krill peptide powder;

4) Add distilled water to the krill peptide powder to make a krill peptide solution, adjust the pH value of the solution to 4-6, add ferrous salt and ascorbic acid to the krill peptide solution, and keep warm at 25-45°C for reaction. After the reaction is over, centrifuge to take the supernatant C to obtain an aqueous solution of krill peptide-ferrous complex;

5) Adjust the pH value of the supernatant C to 6-8, add zinc salt to the supernatant C, and keep warm at 25-45°C for reaction. After the reaction, centrifuge and take the supernatant D to obtain krill peptide-subnatant Iron-zinc complex aqueous solution;

6) Adding calcium salt to the supernatant D, incubating and reacting at 25-45°C, centrifuging after the reaction to take the supernatant E to obtain an aqueous krill peptide-ferrous-zinc-calcium complex solution;

7) The supernatant E was desalted through a nanofiltration membrane, and after freeze-drying, the Antarctic krill peptide-calcium, iron and zinc supplement was obtained.

Wherein the protease described in step (2) is one or more in food-grade papain, bromelain, trypsin, neutral protease or alkaline protease, wherein the best effect with alkaline protease, hydrolysis temperature is 50 ℃ , the pH value of the solution is 8.0, and the hydrolysis time is 2-6h;

Described in step (3) CaCl ₂ addition amount is 1-2% (w/v), nanofiltration membrane molecular weight cut-off is 100Da;

The spray drying conditions described in step (3) are: the solid content in the solution is 8-15%, the air inlet temperature is 130-140°C, and the air outlet temperature is 80-100°C;

The concentration of the krill peptide solution described in step (4) is 1-20%, the ferrous salt is one of ferrous chloride, ferrous sulfate or ammonium ferrous sulfate, and the concentration of ferrous salt is 0.05-0.2mmol / per g krill peptide, the concentration of ascorbic acid is 0.05-0.5% (w/v);

The zinc salt described in step (5) is one of zinc chloride, zinc sulfate or zinc acetate, and the zinc salt concentration is 0.1-0.5mmol/g-krill peptide;

The calcium salt described in step (6) is one of calcium chloride or calcium acetate, and the calcium salt concentration is 3-10mmol/g-krill peptide;

The nanofiltration membrane molecular weight cut-off described in step (7) is 100Da.

The present invention establishes for the first time a process for preparing krill peptide-calcium, iron and zinc supplements using Antarctic krill as a raw material. The raw materials used are abundant and easy to obtain, and have high protein content; Pollution, high product yield, etc., are suitable for industrial production; the prepared krill peptide-calcium iron zinc supplement has the following characteristics: protein content is higher than 90%, product purity is high; fluorine content is lower than 10ppm, product safety is high The molecular weight is mainly distributed in 2000-200Da, with good water solubility and easy absorption by the human body; the content of essential amino acids is high, and the content of calcium, iron and zinc is reasonable. It not only supplements the three nutrients of calcium, iron and zinc, but also supplements essential amino acids. An excellent supplement.

Description of drawings

Figure 1: The molecular weight distribution diagram of the krill peptide prepared by the present invention, wherein the curves with molecular weights of 6512Da, 1355Da and 429Da are respectively the chromatograms of the corresponding molecular weight standards.

Fig. 2: the infrared spectrogram of krill peptide and metal complex thereof of the present invention; Wherein, (a) is the infrared spectrogram of Antarctic krill peptide; (b) is the infrared spectrogram of Antarctic krill peptide-iron complex ; (c) is the infrared spectrum of Antarctic krill peptide-iron-zinc complex; (d) is the infrared spectrum of Antarctic krill peptide-iron-zinc-calcium complex.

Detailed ways

Krill is a group of marine invertebrates that are widely distributed in the world's oceans. According to statistics, the reserves of Antarctic krill are 650-100 million tons, which is a huge treasure house of human protein resources. The protein content of krill dry weight is about 65-75%. Its amino acid composition is reasonable and its essential amino acid content is high. It is an important raw material for high-value utilization of protein. At present, the application and development of Antarctic krill mainly focus on shrimp oil, astaxanthin and chitin, and its protein is mainly used as fishery bait. Using protease to hydrolyze Antarctic krill to prepare low-molecular-weight krill peptides, and using krill peptides as substrates to prepare krill peptide-iron-zinc-calcium composite supplements is one of the effective ways for high-value utilization of krill protein.

The Antarctic krill peptide supplement containing calcium, iron and zinc of the present invention and its preparation method will be described in detail below in conjunction with specific examples.

Example 1

1) Take 10 kg of Antarctic krill powder, add 100 L of 95% ethanol to the krill powder, stir in the reaction kettle for 6 hours, centrifuge to take the precipitate, and dry it in vacuum to obtain defatted krill powder;

2) Take 5kg of defatted shrimp powder, add 60L of distilled water to the defatted shrimp powder, raise the temperature to 50°C, add 2M HCl to the solution to adjust the pH value of the solution to 7.5, add 75g of neutral protease to the solution, hydrolyze at constant temperature for 6h, and heat to 100°C ℃ to terminate the enzyme reaction, and centrifuge to get the supernatant A;

3) Adjust the pH value of the supernatant A to 9.0, add 900g of anhydrous CaCl ₂ to the supernatant A, mix well, let stand at 20°C for 30min, centrifuge to get the supernatant B, add 2M HCl to the supernatant B to adjust The pH value of the supernatant B reached 7.2, and the supernatant B was desalted through a nanofiltration membrane, and spray-dried to obtain krill peptide powder. The molecular weight distribution of the obtained krill peptide powder was mainly 2000-200Da (see Figure 1). In krill peptide powder, the content of essential amino acids is significantly higher than that of krill powder and defatted krill powder (Table 1). It basically conforms to the regulations of FAO/WHO (1985) on the content of essential amino acids in protein required by infants (the contents of valine, histidine, leucine and sulfur-containing amino acids are slightly lower than the regulations).

Table 1: Analysis results of amino acid composition of krill peptide, Antarctic krill meal and defatted Antarctic krill meal prepared by the present invention.

4) Take 2kg of krill peptide powder, add 20L of distilled water to the krill peptide powder to make a krill peptide solution, add 2M HCl to the solution, adjust the pH value of the solution to 5.0, add 68.63g FeCl ₂ 4H ₂ O and 20g ascorbic acid , kept at 25° C. for 2 hours, centrifuged to take supernatant C, which was the krill peptide-ferrous complex aqueous solution. After krill peptide is compounded with ferrous ions, the peak positions of its infrared spectrum bands change significantly (see accompanying drawings 2a and 2b), in which the vibration peak at ^{1455cm disappears} ; The peak position of the vibration peak at ^-1 moves to the lower wave number respectively, that is, 1578cm ^-1 and 1659cm ^-1 , indicating that ferrous ions interact with krill peptide to form a new complex, krill peptide-ferrous Complex.

5) Add 2M NaOH to supernatant C, adjust the pH value of supernatant C to 7.5, add 54.52g ZnCl ₂ to supernatant C, incubate at 25°C for 2 hours, centrifuge to take supernatant D, supernatant D It is krill peptide-ferrous-zinc complex aqueous solution. After the krill peptide-ferrous complex is complexed with zinc ions, the peak positions of some infrared spectrum bands continue to move to the direction of low wavenumbers (see Figure 2b and 2c), among which, the peak positions are located at 1406cm ^-1 , 1630cm ^-1 and ^1659cm -1 The vibration peak at ¹ moved to 1405cm ^-1 , 1629cm ^-1 and 1655cm ^-1 respectively; the vibration peak at 1578cm ^-1 moved to 1590cm ^-1 in the direction of higher wave number, indicating that the middle side of the krill peptide molecule The chain vibration is strengthened, and the hydrogen bond interaction between the molecule and the side chain is weakened; a significant infrared vibration peak appears at 1340cm ^-1 , indicating that the zinc ion interacts with the krill peptide-ferrous complex to form a new complex. That is, krill peptide-ferrous-zinc complex.

6) Add 2.60kg of CaCl ₂ to the supernatant D, incubate at 25°C for 2 hours, centrifuge to get the supernatant E, which is the krill peptide-ferrous-zinc-calcium complex aqueous solution. After the krill peptide-ferrous-zinc complex is combined with calcium ions, the peak positions of some infrared spectrum bands move to the direction of high wave number (see Figure 2c and 2d), among which, the peak positions are located at 1655cm ^-1 , 1405cm ^-1 and 1340cm The peak position of the vibration peak at ^-1 moved to ^{1661cm -1 , 1414cm -1 and 1343cm -1} respectively; The shrimp peptide-ferrous-zinc complex interacts to form a new complex, namely the krill peptide-ferrous-zinc-calcium complex.

7) The supernatant E was desalted through a nanofiltration membrane and freeze-dried to obtain the Antarctic krill peptide-calcium, iron and zinc supplement.

In the Antarctic krill peptide-calcium-iron-zinc supplement prepared by the present invention, the iron content is 2.40mg-Fe/g-product, the zinc content is 8.28mg-Zn/g-product, and the calcium content is 145.38mg-Ca/g-product . According to the dietary guidelines for Chinese residents on the daily recommended intake of nutrients: Infants and young children, the recommended daily intake of calcium is 600-800mg/g, the recommended daily intake of zinc is 9-12mg/g, and the daily recommended intake of iron The recommended intake is 10-12mg/g; the recommended daily intake of calcium for adults is 800mg/g, the recommended daily intake of zinc is 11.5-15mg/g, and the recommended daily intake of iron is 15-20mg/g g. The supplement prepared by the invention can reasonably supplement three nutrients and improve human health.

Example 2

(1) Take 10 kg of Antarctic krill powder, add 80 L of 95% ethanol to the krill powder, stir in the reaction kettle for 7 hours, centrifuge to get the precipitate, and vacuum-dry to obtain defatted krill powder;

(2) Take 5kg of defatted shrimp powder, add 50L of distilled water to the defatted shrimp powder, add 2M HCl to the solution, adjust the pH value of the solution to 8.0, heat up to 50°C, add 100g of alkaline protease, hydrolyze at constant temperature for 4 hours, and heat to 100°C Terminate the enzyme reaction, centrifuge to get the supernatant A;

(3) Adjust the pH value of the supernatant A to 8.5, add 600g of anhydrous CaCl ₂ to the supernatant A, mix well, let stand at 20°C for 30min, centrifuge to get the supernatant B, add 2M HCl to the supernatant B , adjusting the pH value of the supernatant B to 7.0-7.5, desalting the supernatant B through a nanofiltration membrane, and spray-drying to obtain krill peptide powder. The molecular weight distribution of the obtained krill peptide powder is mainly 2000-200Da.

(4) Take 2 kg of krill peptide, add 30 L of distilled water to the krill peptide to make a krill peptide solution, add 2M HCl to the solution, adjust the pH value of the solution to 4.5, add 79.50 g FeCl ₂ 4H ₂ O and 25 g of ascorbic acid, Incubate at 37°C for 2 hours, centrifuge to take supernatant C, supernatant C is krill peptide-ferrous complex aqueous solution;

(5) Add 2M NaOH to the supernatant C, adjust the pH value of the supernatant C to 7.0, add 40.89g ZnCl ₂ to the solution, incubate at 37°C for 2 hours, centrifuge to take the supernatant D, and the supernatant D is It is an aqueous solution of krill peptide-ferrous-zinc complex;

(6) Add 2kg of CaCl ₂ to supernatant D, incubate at 37°C for 2 hours, centrifuge to get supernatant E, supernatant E is krill peptide-ferrous-zinc-calcium complex aqueous solution;

(7) The supernatant E was desalted by nanofiltration membrane and freeze-dried to obtain Antarctic krill peptide-calcium iron zinc supplement.

After testing, in the Antarctic krill peptide-calcium iron zinc supplement, the iron content is 2.78mg-Fe/g-product, the zinc content is 8.35mg-Zn/g-product, and the calcium content is 137.64mg-Ca/g-product .

The Antarctic krill peptide-calcium iron zinc supplement prepared by the present invention has the following characteristics: the protein content is higher than 90%, and the product purity is high; the fluorine content is lower than 10ppm, and the product safety is high; It has good properties and is easy to be absorbed by the human body; the content of essential amino acids is high, and the content of calcium, iron and zinc is reasonable. It not only supplements the three nutrients of calcium, iron and zinc, but also supplements essential amino acids. It is an excellent supplement.

Claims (7)

1. the krill peptide replenishers containing calcium iron zinc, it is characterized in that, described replenishers prepare by following method:

1) by euphausia superba powder after Ethanol Treatment, drying and crushing obtains degreasing krill meal;

2) in degreasing krill meal, add distilled water, feed liquid mass ratio is 1:8 ~ 12, and regulate solution ph to be 7.0 ~ 8.0, then in solution, add protease be hydrolyzed, hydrolysis temperature is 50 DEG C, and solution ph is 8.0, and hydrolysis time is 2-6h; Hydrolysis terminates rear heating termination enzyme digestion reaction, centrifuging and taking supernatant A;

3) pH value of supernatant A is adjusted to 8.5 ~ 9.5, in supernatant A, adds CaCl ₂defluorinate, leave standstill after mixing, centrifuging and taking supernatant B, by supernatant B adjust ph to 7.0 ~ 7.5, supernatant B crosses NF membrane desalination, spray-dried krill Gly-His-Lys;

4) in krill Gly-His-Lys, add distilled water and be made into krill peptide solution, solution ph is adjusted to 4 ~ 6, in krill peptide solution, add ferrous salt and ascorbic acid, insulation reaction under 25-45 DEG C of condition, reaction terminates rear centrifuging and taking supernatant C and obtains krill peptide-ferrous compound water solution;

Wherein ferrous salt is any one or several in frerrous chloride, ferrous sulfate or iron ammonium sulfate, and ferrous salt and the proportionate relationship of krill peptide are the corresponding 0.05-0.2mmol ferrous salt of every g krill peptide;

5) pH value of supernatant C is adjusted to 6 ~ 8, in supernatant C, adds zinc salt, insulation reaction under 25-45 DEG C of condition, reaction terminates rear centrifuging and taking supernatant D and obtains krill peptide-ferrous iron-zinc complexes aqueous solution; Wherein zinc salt is any one or several in zinc chloride, zinc sulfate or zinc acetate, and zinc salt and the proportionate relationship of krill peptide are the corresponding 0.1-0.5mmol zinc salt of every g krill peptide;

6) in supernatant D, add calcium salt, insulation reaction under 25-45 DEG C of condition, reaction terminates rear centrifuging and taking supernatant E and obtains krill peptide-ferrous iron-zinc-calcium complexes aqueous solution; Wherein calcium salt is any one or two kinds of in calcium chloride or calcium acetate, and calcium salt and the proportionate relationship of krill peptide are the corresponding 3-10mmol calcium salt of every g krill peptide;

7) supernatant E is crossed NF membrane desalination, after freeze drying, obtain krill peptide-calcium iron Zinc supplements.

2. krill peptide replenishers as claimed in claim 1, is characterized in that, described step 2) in protease be one or more in food-grade papain, bromelain, trypsase, neutral proteinase or alkali protease.

3. krill peptide replenishers as claimed in claim 2, is characterized in that, described step 2) in protease be alkali protease.

4. krill peptide replenishers as claimed in claim 1, is characterized in that, described step 3) middle CaCl ₂addition be 1-2%.

5. krill peptide replenishers as claimed in claim 1, is characterized in that, described step 3) in spray drying condition as follows: in solution, solid content is 8-15%, and EAT is 130-140 DEG C, and leaving air temp is 80-100 DEG C.

6. krill peptide replenishers as claimed in claim 1, is characterized in that, described step 4) in the concentration of krill peptide solution be 1-20%.

7. krill peptide replenishers as claimed in claim 1, is characterized in that, described step 4) in ascorbic acid concentrations be 0.05-0.5%.