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Introduction Calcium plays an important role in the body, such as acting as a cell messenger for signaling, coagulation, muscle contraction, and the formation of the body
.
Therefore, calcium deficiency can lead to metabolic bone diseases, etc.
, which is harmful to health
.
Dietary intake is the only way to replenish calcium in the body, however, ionized calcium tends to form insoluble complexes in the alkaline environment of the intestine or in the presence of other dietary components such as oxalic acid and phytic acid, resulting in insufficient
calcium absorption and utilization.
Studies have shown that casein phosphopeptides (CPPs) can bind calcium ions to form soluble peptide-calcium complexes, promote calcium absorption, and improve the accumulation
of calcium in bone.
CPPs produced by casein can promote calcium transport by upregulating the TRPV6 key calcium transporter expression in Caco-2 cells, which can promote the expression
of TRPV6 in rat intestinal cells.
Yolk high phosphoprotein phosphopeptide, produced by enzymatic hydrolysis of yolk high phosphoprotein in egg yolk granules, has a similar structure to casein phosphopeptide, and has a higher phosphorylation level
than casein.
Yolk high phosphoprotein (Phosvitin, PV) is a natural phosphorylated protein in the yolk, accounting for 8.
0%~11% of the yolk protein, composed of 217 amino acids, of which 128 amino acid residues can be phosphorylated
.
PV phosphorylated serine residues are arranged in succession to form serine clusters, which makes it difficult
to deeply enzymatic hydrolysis of PV.
Many studies use partial dephosphorylation and enzymatic hydrolysis with some success, but the phosphate group
is lost.
Our team developed a high-temperature and medium-pressure combined enzymatic hydrolysis technology, which greatly improved the enzymatic hydrolysis effect of PV, increased the number of PPP (HTMP-Enz-PPP), and produced up to 605 phosphopeptides
from PV.
Yolk high phosphoprotein phosphopeptide has similar structural functions to casein phosphopeptide, and it is speculated that it can also promote calcium absorption and deposition in vivo, so this paper studies the effect
of yolk high phosphoprotein phosphopeptide-calcium complex on calcium absorption and bone deposition in mice.
Results and Discussion When the mass ratio of peptide to calcium reached 7:1, the calcium binding rate increased to 98.
44%, indicating that almost all the added calcium formed PPP-Ca
complexes.
The Zeta potential values in the sample are all negatively charged, indicating that the surfaces of PV and PPP are mainly negatively
charged.
When calcium binds, calcium ions neutralize the negative charge on its surface, and the Zeta potential of the complex rises
.
Fig.
1.
Preparation and structural characterization of peptide-calcium chelate The digestive stability
of PV-Ca and PPP-Ca complexes to pepsin and trypsin was evaluated.
After 30 min of incubation in simulated gastric juice, the calcium retention rate of PV-Ca was 35.
48%, which was relatively low and did not change much
within 6 h of incubation.
HTMP-Enz-PPP formed after enzymatic digestion of PV has better calcium-holding ability, and the calcium retention rate in simulated gastric juice is 65%~72%, and there is no significant change
in 6 h incubation time.
Fig.
2 Effect of simulated gastrointestinal (stomach with pH 2.
0) digestion on the stability of protein/peptide calcium complex The bone density of mice in the low calcium control group was 0.
17 g/cm3, which was significantly lower than that of the normal control group (0.
29 g/cm3).
After 4 weeks of calcium supplementation, the bone density of mice fed calcium chloride was only 0.
19 g/cm3, while that of PV-Ca group and HTMP-Enz-PPP-Ca group was 0.
23 and 0.
27 g/cm3
, respectively.
Calcium deposition and bone density results showed that HTMP-Enz-PPP-Ca had better effects on osteogenic quality than inorganic calcium
during bone remineralization.
Micro-CT images of the distal femur showed that the bone tissue in the middle of the femur was loose in the low calcium control group and calcium chloride mice, and the trabecular bone structure was severely damaged
.
The results showed that HTMP-Enz-PPP-Ca significantly improved the microstructure of bone tissue and improved the integrity
of bone trabeculae.
A.
Normal group; B.
Low calcium control; C.
PV-CaCl2； D.
HTMP-Enz-PPP-CaCl2； E.
CaCl2 Figure 3 Conclusion of distal femur 3D micro-CT images
of each group Conclusion prepared PPP-Ca complexes by high-temperature medium pressure-enzymatic hydrolysis treatment of PV, measured their stability in simulated gastrointestinal tract, established a low-calcium SPF-KM mouse model, and evaluated the effect of
PPP-Ca complexes on bone formation 。 HTMP-Enz-PPP-Ca feeding significantly reduced the abnormal elevation of serum alkaline phosphatase (AKP) caused by calcium deficiency, and the serum calcium content was the highest, significantly improving bone calcium content and bone mineral density
.
The femur of HTMP-Enz-PPP-Ca intervention had a more compact, complete and dense trabecular bone network structure
than the low calcium and calcium chloride groups.
These results showed that egg yolk phosphopeptide calcium complex could effectively promote calcium absorption and bone deposition, and the effect was better than inorganic calcium
chloride.
First author
Mengdie Zhao, female, holds a master's degree
from the College of Food Science and Technology, Huazhong Agricultural University.
His main research direction is the preparation of yolk high phosphoprotein phosphopeptide calcium complex and its calcium-promoting absorption mechanism
.
Huang Qian, professor of Huazhong Agricultural University, doctoral supervisor, director of the Department of Food Nutrition and Health, visiting scholar at Iowa State University, has successively served as the deputy secretary-general and director of the China Animal Products Processing Research Association of the national first-level society, a member of the Youth Committee, and a director of
the Asian Egg Association.
The winner of the "Youth Science and Technology Morning Light Program" in Hubei Province, he was awarded the title of "Outstanding Youth of China's Egg Processing Industry", and the title of Class A Young Talent of the "Shishan Shuoyan Program" of Huazhong Agricultural University
.
He has presided over more than 10 projects such as 3 projects of the National Natural Science Foundation of China, key research and development projects of the National 13th Five-Year Plan, special sub-projects of public welfare industries, and projects of the Hubei Provincial Natural Science Foundation
.
He has published more than 40 SCI/EI papers
.
Associate editor of the "Twelfth Five-Year Plan" textbook "Animal Food Processing" of the Ministry of Agriculture, participated in the compilation of 1 English monograph of the Royal Society of Chemistry and 2 planning textbooks of the Ministry of Agriculture
.
It has obtained 11 invention patents authorized by the state, including 5 international patents in the United States, Japan and Australia, and 1 achievement transformation
.
Won the first and second prizes of Hubei Province Science and Technology Progress Award
.
In 2021, he received the American Egg Board Research Award from the American Poultry Science Association (PSA
).
Food Hydrocolloids、Anal.
Chim.
Acta、Biosens.
Bioelectron.
、Int.
J.
Bio.
Reviewers
for SCI magazines such as Macr.
and Poultry Sci.
Mengdie Zhaoa, Dong Uk Ahnb, Songming Lia, Wei Liua, Shengwei Yic, Xi Huanga,*
a National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China
b Animal Science Department, Iowa State University, Ames 50011, USA
c Chongqing Institute for Food and Drug Control, Chongqing 401121, China
*Corresponding author.
E-mail address: huangxi@mail.
hzau.
edu.
cn
Phosvitin (PV) was treated with high-temperature, mild pressure (HTMP), and enzyme combination, and then phosvitin phosphopeptides-calcium (PPP-Ca) complexes were prepared.
The low-calcium specific pathogen free-Kunming (SPF-KM) mice were used to determine the effect of PPP-Ca complexes on intestinal calcium absorption and their utilization for bone formation.
The serum calcium content was the highest with the HTMP-Enz-PPP-Ca treatment (2.
19 mmol/L), and it significantly down-regulated the abnormal elevation of serum alkaline phosphatase (AKP) caused by calcium deficiency.
The low-calcium control group had the lowest calcium deposited to the femur (80.
41 mg/g) and the lowest femur bone mineral density (BMD) (0.
17 g/cm3 ), while HTMP-Enz-PPP-Ca significantly improved bone calcium content (94.
33 mg/g) and BMD (0.
29 g/cm3 ).
The micro-computed tomography (MCT) images showed that the femur with the normal control, PV-Ca, and HTMP-Enz-PPP-Ca treatments had a more compact, complete, and thicker trabecular network than the low-calcium and CaCl2 treatments.
These results indicated that the organic calcium (HTMP-Enz-PPP-Ca) promoted calcium absorption and bone deposition, and the effect of HTMP-Enz-PPP-Ca was better than the inorganic CaCl2.
Reference:
ZHAO M D, AHN D U, LI S M, et al.
Effects of phosvitin phosphopeptide-Ca complex prepared by efficient enzymatic hydrolysis on calcium absorption and bone deposition of mice[J].
Food Science and Human Wellness, 2022, 11(6): 1631-1640.
DOI:10.
1016/j.
fshw.
2022.
06.
022.
