Fabrication of Human Milk Fat Substitute: Based on the Similarity Evaluation Model and Computer Software.

We aimed to obtain the optimal formula for human milk fat substitute (HMFS) through a combination of software and an evaluation model and further verify its practicability through an animal experiment. The results showed that a total of 33 fatty acid (FA) and 63 triglyceride (TAG) molecular species were detected in vegetable oils. Palmitic acid, oleic acid, linoleic acid, 18:1/16:0/18:1, 18:2/16:0/18:2, 18:1/18:1/18:1 and 18:1/18:2/18:1, were the main molecular species among the FAs and TAGs in the vegetable oils. Based on the HMFS evaluation model, the optimal mixed vegetable oil formula was blended with 21.3% palm oil, 2.8% linseed oil, 2.6% soybean oil, 29.9% rapeseed oil and 43.4% maize oil, with the highest score of 83.146. Moreover, there was no difference in the weight, blood routine indices or calcium and magnesium concentrations in the feces of the mice between the homemade mixed vegetable oil (HMVO) group and the commercial mixed vegetable oil (CMVO) group, while nervonic acid (C24:1) and octanoic acid (C8:0) were absorbed easily in the HMVO group. Therefore, these results demonstrate that the mixing of the different vegetable oils was feasible via a combination of computer software and an evaluation model and provided a new way to produce HMFS.

Effects of endogenous DHA milk and exogenous DHA milk on oxidative stress and cognition in SAMP8 mice.

In this study, Senescence Accelerated Mice (SAMP8) were supplemented with exogenous DHA milk, endogenous DHA milk, normal milk, or 0.9 % saline solution. Enzyme-linked immunosorbent assay (ELISA), gas chromatography (GC), ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI MS/MS), and Morris water maze were used to characterize the effects of diet on oxidative stress and cognition in SAMP8 mice. Supplementation endogenous DHA milk or exogenous DHA milk can enhance the antioxidant capacity of mice organs. Endogenous DHA milk increased the superoxide dismutase (SOD) activity of mice brain and serum than normal milk and 0.9 % saline solution (P ≤ 0.05), as well as increased SOD activity of mice liver and glutathione peroxidase (GSH-Px) activity of mice brain than normal milk (P ≤ 0.05). Exogenous DHA milk increased SOD activity of mice brain than normal milk and 0.9 % saline solution, as well as increased SOD activity of mice serum than 0.9 % saline solution (P ≤ 0.05). Several polar lipid relative content, such as 18:0/18:2 PS, 17:0 Ceramide, and 20:4 LPC in mice brain was affected by dietary supplementation with DHA-containing milk. Lipid oxidation metabolites in mice brain were not affected by DHA-containing milk. Endogenous DHA milk increased the number of platform location crossing times of mice in the Morris water maze test, compared with Exogenous DHA milk, normal milk, and 0.9 % saline solution (P ≤ 0.05).

Characteristics of casein phosphopeptides in Chinese human milk and its correlation with infant growth: A cross-sectional study.

This research aimed to investigate the characteristics of casein phosphopeptides in Chinese human milk, and their potential relationship to infant growth. Using the liquid chromatography-Orbitrap-mass spectrometry technique, a total of 15 casein phosphopeptides were identified from 200 human milk samples. Also, our results indicate that casein phosphopeptides were phosphorylated with only one phosphate. The relative concentrations of casein phosphopeptides at 6 months postpartum were increased compared with milk at 2 months (FDR < 0.05). Significantly positive correlations were observed between casein phosphopeptides and infant growth, as shown by four casein phosphopeptides were positively correlated with the infants' weight-for-age Z-scores (rs range from 0.20 to 0.29), and three casein phosphopeptides were positively correlated with the infants' length-for-age Z-scores (rs range from 0.19 to 0.27). This study is the first to reveal the phosphorylated level and composition of casein phosphopeptides in Chinese human milk, and their potential relationship with infant growth.

Effects of Pre-Emulsification with Thermal-Denatured Whey Protein on Texture and Microstructure of Reduced-Sodium Processed Cheese.

Thermal-denatured whey protein-milk fat emulsion gels with different degrees of pre-emulsification were prepared by pre-emulsifying milk fat with thermal-denatured whey protein and used in the preparation of reduced-sodium processed cheeses. The effect of the thermal-denatured whey protein pre-emulsification process on the texture and microstructure of reduced-sodium processed cheeses was evaluated by studying the composition, color, texture, functional properties, microstructure and sensory analysis of the processed cheeses. The results showed that compared with cheese without pre-emulsified fat (1.5% ES control), the moisture content of cheese with pre-emulsified 100% fat (1.5% ES100) increased by 5.81%, the L* values increased by 7.61%, the hardness increased by 43.24%, and the free oil release decreased by 38%. The microstructure showed that the particle size of fat was significantly reduced, and the distribution was more uniform. In addition, compared with the cheese added with 3% emulsifying salt (3% ES control), the amount of emulsifying salt in the 1.5% ES100 decreased by 50%, but the fat distribution of the two kinds of cheese tended to be consistent, and there was no obvious change in texture characteristics and meltability. Sensory scores increased with the increase in pre-emulsification degree. Overall, the pre-emulsification of milk fat with thermal-denatured whey protein can reduce the sodium content of processed cheese and improve its quality.

Changes in glycosylated proteins in colostrum and mature milk and their implication.

Introduction: Glycosylation is one of the essential post-translational modifications that influences the function of milk proteins. Methods: In the present study, 998 proteins and 764 glycosylated sites from 402 glycoproteins were identified in human milk by TMT labeling proteomics. Compared to human milk proteins, the glycoproteins were mainly enriched in cell adhesion, proteolysis, and defense/immune process. Results: The abundance of 353 glycosylated sites and their 179 parent proteins was quantified. After normalization to their parent protein's abundance, 78 glycosylated sites in 56 glycoproteins and 10 glycosylated sites in 10 glycoproteins were significantly higher in colostrum and mature milk, respectively. These changed glycoproteins were mainly related to host defense. Intriguingly, one glycosylated site (Asp144) in IgA and two glycosylated sites (Asp38 and Asp1079) in tenascin are significantly upregulated even though their protein abundance was downregulated during lactation. Discussion: This study helps us figure out the critical glycosylated sites in proteins that might influence their biological function in an unbiased way.

Determination and Risk Assessment of Flavor Components in Flavored Milk.

This study aimed to determine chemical composition and assess exposure in flavored milk among Chinese residents, based on risk assessment methodologies of acceptable daily intake (ADI) and toxicological concern threshold (TTC). Esters (32.17%), alcohols (11.19%), olefins (9.09%), aldehydes (8.39%), and ketones (7.34%) comprised the majority of the flavoring samples. Methyl palmitate (90.91%), ethyl butyrate (81.82%), and dipentene (81.82%) had the highest detection rates in flavor samples. This study screened fifteen flavor components of concern and discovered that 2,3,5-trimethylpyrazine, furfural, benzaldehyde, and benzenemethanol were detected in 100% of flavored milk samples. Benzenemethanol was found in the highest concentration (14,995.44 µg kg-1). The risk assessment results revealed that there was no risk for Chinese residents in consuming flavored milk, and the maximum per capita daily consumption of 2,3,5-trimethylpyrazine, furfural, and benzenemethanol were 226.208 g, 140.610 g, and 120.036 g, respectively. This study could provide guidelines for amounts of flavor additive ingredients in milk.

Advances in the Metabolic Mechanism and Functional Characteristics of Equol.

Equol is the most potent soy isoflavone metabolite and is produced by specific intestinal microorganisms of mammals. It has promising application possibilities for preventing chronic diseases such as cardiovascular disease, breast cancer, and prostate cancer due to its high antioxidant activity and hormone-like activity. Thus, it is of great significance to systematically study the efficient preparation method of equol and its functional activity. This paper elaborates on the metabolic mechanism of equol in humans; focuses on the biological characteristics, synthesis methods, and the currently isolated equol-producing bacteria; and looks forward to its future development and application direction, aiming to provide guidance for the application and promotion of equol in the field of food and health products.

Fatty acid, triglyceride, and kinetic properties of milk fat fractions made by the combination of dry fractionation and short-path molecular distillation.

In this study, we aimed to detect the physicochemical properties of distilled products (residue and distillate) obtained from anhydrous milk fat (AMF) and its dry fractionation products (liquid and solid fractions at 25°C [25 L and 25 S]). The results showed that the saturated fatty acids and low- and medium molecular-weight triglycerides were easily accumulated in the distillate, and the percentage of unsaturated fatty acid and high molecular-weight triglycerides in the residue were higher, and these components in 25 S and 25 L were influenced more significantly than those in the AMF. In addition, the distillate had larger melting ranges in comparison with the distilled substrate, while the melting ranges of residue was smaller. The triglycerides were presented as the mixture crystal forms (α, ß', and ß crystal) in 25 S, AMF, and their distilling products, and it was transformed gradually to a single form as the increasing of distilling temperature. Moreover, the accumulated pattern of triglycerides was double chain length in 25 S, AMF, and their distilling products. These results provide a new approach to obtain the milk fat fractions with different properties, and the findings of this study enrich the theoretical basis of milk fat separation in practical production.

Effects of different heat treatments on Maillard reaction products and volatile substances of camel milk.

Camel milk has unique compositional, functional and therapeutic properties compared to cow's milk and also contains many protective proteins with anti-cancer, anti-diabetic and anti-bacterial properties. In this experiment, fresh camel milk was heat-treated at different temperatures and times, and the changes in Millard reaction products were analyzed. Meanwhile, headspace-gas chromatography-ion migration spectrometry (HS-GC-IMS), electronic nose and electronic tongue were used to analyze the changes of volatile components in camel milk after different heat treatments. The results showed that the Maillard reaction was more severe with the increase of heat treatment, and the contents of furosine and 5-hydroxymethylfurfural increased significantly when the heat treatment temperature was higher than 120°C. HS-GC-IMS results showed that the contents of aldehydes and ketones increased obviously with the increase of heat treatment degree. The study clarifies the effects of different heat treatment degrees on Maillard reaction degree and flavor of camel milk, which has practical production guidance significance for the research and industrialization of liquid camel milk products.

Comparison of glycerophospholipid and sphingolipid in mature milk from different sampled regions in the Chinese human milk project (CHMP) study.

Milk phospholipids (PLs) are critical components of infant growth. This study aimed to discover PL in mature human milk (HM) from China (n = 201) and mainly assessed the effect caused by sampled regions. The average total PL concentration was quantified from 3.65 to 11.25 mg per g of lipid, and the major PL class identified was sphingomyelin (SM, 38.06-47.62 %), followed by phosphatidylcholine (PC, 29.61-34.39 %), and phosphatidylethanolamine (PE, 10.54-24.46 %). In addition, the 36:2 (18:0/18:2), 38:6 (16:0/22:6), 40:1 (d18:1/22:0), and 42:2 (d18:1/24:1) were the most abundant molecular species identified in glycerophospholipid and SM molecular species respectively. Some PL molecular species were strongly related with region of sampling, like lysophosphatidylinositol 18:1 was only detected in Beijing. In conclusion, those findings showed that the PL molecular species and concentration of HM had significant regional diversity, and it will give the Chinese human milk database more accurate PL data.

Dietary Schizochytrium Microalgae Affect the Fatty Acid Profile of Goat Milk: Quantification of Docosahexaenoic Acid (DHA) and Its Distribution at Sn-2 Position.

The objective of this study was to detect the influence of dietary Schizochytrium microalgae on milk composition, milk fatty acids, and milk sn-2 fatty acids in goat's milk. Firstly, we could see that the fat content increased in low microalgae supplementation goat's milk (LM, 15 g/day) and the lactose content decreased in medium microalgae supplementation goat's milk (MM, 25 g/day) compared with control goat's milk (C, 0 g/day). Moreover, the absolute concentration of the docosahexaenoic acid (DHA) of LM, MM, and high microalgae supplementation (HM, 35 g/day) goat's milk was 29.485, 32.351, and 24.817 mg/100 g raw milk, respectively, which were all higher than that in the control goat's milk with 4.668 mg/100 g raw milk. In addition, the sn-2 DHA content increased in MM and HM goat's milk. However, the decreasing trend of the sn-2 DHA content was observed in LM goat's milk. As for other fatty acids, the oleic acid (C18:1n9c) and linolenic acid (C18:3n3) content decreased and increased, respectively, in all experimental goat milk. Finally, an interesting phenomenon was found, which was that docosanoic acid (C22:0) and tetracosenic acid (C24:1) were only detected in test goat's milk. Consequently, the phenomena of this study demonstrated that dietary Schizochytrium microalgae have an obvious effect on the fatty acid and sn-2 fatty acid profile of goat's milk, and they provide an effective method to improve the content of goat's milk DHA in practical production.

Effects of Monascus on Proteolysis, Lipolysis, and Volatile Compounds of Camembert-Type Cheese during Ripening.

In order to improve the flavor and taste of Camembert cheese, the use of Monascus as an adjunct starter for the production of Camembert-type cheese was studied to investigate its effect on the proteolysis, lipolysis, and volatile compounds during ripening for 40 days. The Camembert cheese without Monascus was used as a control. The results showed that proteolytic and lipolytic activities increased to a certain extent. The addition of Monascus promoted primary and secondary proteolysis, due to the release of some proteases by Monascus. Aspartic, Threonine, Glutamic, Glycine, Methione, Isoleucine, Phenyalanine, and Lysine contents in experimental group (R) cheese were significantly higher than those in control group (W) cheeses. In addition, the free amino acid and fatty acid contents were also affected. The identification of flavor components using gas-mass spectrometry (GC-MS) showed that 2-undecone, 2-tridecanone, phenylethyl alcohol, butanediol (responsible for the production of flowery and honey-like aroma), ethyl hexanoate, ethyl octanoate, and ethyl citrate (fruit-like aroma) were significantly higher (p < 0.05) in the experimental cheeses than in the control. The contents of 2-nonanone, 2-octanone and 2-decanone (showing milky flavor), and 1-octene-3 alcohol with typical mushroom-like flavor were lower than the control.

Ultrasound pretreatment prior to spray drying improve the flowability and water sorption properties of micellar casein concentrate.

This research investigated the effect of ultrasound (US) pretreatment prior to spray drying on the powder flow and moisture sorption behaviour of micellar casein concentrate (MCC). MCC produced from skim milk microfiltration was sonicated at energy intensity of 0 (control), 47 J/mL (S-2000), 62 J/mL (S-3000) and 76 J/mL (S-4000). The results revealed that US pretreatment significantly increased the average particle size (D50) from 82.46 µm to 100.73 µm and reduced the surface fat content from 19.2% to 13.8%, resulting in decreased basic flow energy, cake energy and cohesion. Besides, the US treated samples showed relatively poor ability to acquire the moisture from the atmosphere than the control. Protein structure analysis showed that α-helix decreased with enhanced US power, while ß-sheet and surface hydrophobicity increased, implying hydrophobic groups were exposed and water sorption rate was impeded. As a result, US pretreatment can improve the powder flow and potentially reduce the negative effect of cake formation at high humidity.

Chemical and Flavor Characteristics of Enzyme-Modified Cheese Made by Two-Stage Processing.

(1) Background: to date, a clear description of the impact of specific enzymes on the enzyme-modified cheese (EMC) flavor is lacking. Moreover, comparative studies on the aroma compounds' intensity of EMC have been rarely investigated. Therefore, this study was done to determine the influence of incubating substrates with proteases and different lipases on cheese ripening index and aroma compounds. (2) Methods: two-stage processing was adopted; proteolysis followed by lipolysis. (3) Results: results showed that the usage of Flavourzyme may improve the value of pH 4.6-WSN/TN%. Butanoic acid and hexanoic acid have a significant influence on the overall flavor of EMCs. In particular, the ethenyl acetate compound was detected in all products and was perceived as a fruity and sweet aroma, which has not been reported in previous literature. The concentration of short-chain fatty acids of EMCs made by Lipase MER was higher than EMCs made by Palatase, while the total content of medium and long-chain fatty acids of EMCs made by Lipase MER was lower than EMCs made by Palatase. The percentage of esters compounds in EMCs made by Lipase AY 30G was higher than the other two lipases, except EMC1. (4) Conclusions: Flavourzyme may be used to speed up the ripening of cheeses that need extensive proteolysis. The ability of Lipase MER to hydrolyze short-chain fatty acids was stronger than that of Palatase, while the ability of Lipase MER to hydrolyze medium and long-chain fatty acids was weaker than that of Palatase. The use of Lipase AY 30G was accompanied by the production of some other flavor esters, which made the final hydrolysates more fragrant and may be a good choice to produce fruity cheese flavor EMC. While Lipase MER may barely contain ester activity. This study may provide a reference for the selection of incubated enzymes for specific flavor EMC.

Effect of Different Temperature-Controlled Ultrasound on the Physical and Functional Properties of Micellar Casein Concentrate.

Micellar casein concentrate (MCC) is a novel dairy ingredient with high protein content. However, its poor functional properties impair its potential for further application, highlighting the importance of using innovative processing methods to produce modified MCC, such as ultrasound (US). This work investigated the impact of US on the physical and functional properties of MCC under temperature-controlled and -uncontrolled conditions for different time intervals. Under temperature-controlled ultrasound (TC-US) treatment, a reduction was found in the supernatant particle size of casein micelles. Soluble calcium content and hydrophobicity increased following ultrasound treatment at 20 °C, resulting in a remarkable improvement in emulsification. However, long-time ultrasonication led to an unstable state, causing the MCC solutions to show shear thinning behavior (pseudoplastic fluid). Compared with 50 °C temperature-controlled ultrasonication, ultrasonication at 20 °C had a greater influence on particle size, viscosity and hydrophobicity. These findings indicate that 20 °C TC-US could be a promising technology for the modification of MCC.

Comparative Proteomics of Human Milk From Eight Cities in China During Six Months of Lactation in the Chinese Human Milk Project Study.

Human milk (HM) is the golden standard of infant nutrition that can protect immature body function and enhance nutrition metabolism to ensure infant growth. Region specificity and lactation period could change the protein composition in HM. In this research, proteomics analysis was used to compare proteomes across eight cities, namely Harbin, Lanzhou, Guangzhou, Chengdu, Jinhua, Weihai, Zhengzhou, and Beijing, which represented the northeast, northwest, southeast, southwest, east, and north and central regions of China,. Proteins varied significantly among the cities. These different proteins were mainly involved in the process of platelet degranulation, innate immune response, and triglyceride metabolic process, which might be due to different living environments. These differences also lead to variation in protection and fat metabolism from mothers to infants in different cities. Four proteins were expressed differently during 6 months of lactation, namely Dipeptidyl peptidase 1, Lysozyme C, Carbonic anhydrase 6, and Chordin-like protein 2. The changes in these proteins might be because of the change of growth needs of the infants. The findings from our results might help to improve the understanding of HM as well as to design infant formula.

Comprehensive Human Milk Patterns Are Related to Infant Growth and Allergy in the CHMP Study.

SCOPE: The aim of the present study is to identify human milk pattern using multi-omics datasets and to explore association between patterns, infant growth, and allergy using data from the Chinese Human Milk Project (CHMP) study. METHODS AND RESULTS: Three patterns are identified from integrative analysis of proteome, lipidome, and glycome profiles of 143 mature human milk samples. Factor 1 is positively associated with 128 proteins, phospholipids, and human milk oligosaccharides (HMOs) including lacto N-neohexaose (LNnH) and lacto-N-difucohexaose II (LNDFH II); factor 2 is negatively associated with as1 -casein, phospholipids while positively associates with HMOs including LNnH, lactosialyl tetrasaccharide c (LSTc), and 2'-fucosyllactose (2'FL); factor 3 is positively associated with lysophospholipids while negatively associates with 27 proteins, triglycerides with two saturated fatty acids, 6'-sialyllactose (6'SL) and 2'FL. In general, factor 1 and factor 2 are associated with slower while factor 3 is associated with faster growth rate (p < 0.044). One unit higher in loadings of factor 2 is associated with 34% lower risk of allergies (p ≤ 0.017). Associations are not significant after adjustment for city except for factor 1. CONCLUSIONS: Three possible human milk patterns with varying degree of stability are identified. Future work is needed to understand these patterns in terms of generalization, biologic mechanisms, and genotype influences.

Preparation and drying of water-in-oil-in-water (W/O/W) double emulsion to encapsulate soy peptides.

Soy peptide solution (40%, w/w) was successfully encapsulated in a W1/O/W2 double emulsion produced by a two-step emulsification process. Polyglycerol polyricinoleate (PGPR) was found to be an effective inner emulsifier compared to Span 60 and lecithin to produce stable W1/O primary emulsion. The primary emulsion was subsequently emulsified into an outer aqueous phase (W2) containing octenyl succinic anhydride (OSA) starch and maltodextrin. The droplet size and encapsulation efficiency of the peptide solution in W1/O/W2 emulsion were found to depend on the W1:O ratio, peptide concentration in the inner W1 phase and homogenization condition of the secondary emulsification step. The double emulsion with the highest encapsulation efficiency (>80%) was prepared by: (i) using 40% (w/w) soy peptide solution as W1 phase; (ii) controlling W1:O ratio at 3:7 (w/w) and (iii) homogenizing the emulsion at 10,000 rpm for 3 min. The freeze-dried microcapsule powder of W1/O/W2 emulsion showed higher encapsulation efficiency (>70%) compared to spray-dried one. The freeze-dried microcapsule of W1/O/W2 double emulsion developed in this study is a promising delivery matrix to encapsulate hydrophilic ingredients including peptides. Fourier-transform infrared spectroscopy (FTIR) spectra of the microcapsule powder indicated good compatibility between peptide and encapsulants.

Comparative Analysis of Triglycerides From Different Regions and Mature Lactation Periods in Chinese Human Milk Project (CHMP) Study.

The kinds and proportions of triglycerides of human mature milk play an independent role in the growth of infants. In this study, the human milk samples obtained from eight different Chinese cities (Chengdu, Weihai, Lanzhou, Jinhua, Beijing, Guangzhou, Zhengzhou, and Harbin) and six sequential mature lactation times (30, 60, 90, 120, 150, and 180 days) were detected for the triglycerides. The result demonstrated that total 66 triglycerides were detected in mature human milk, with acyl carbon number (ACN) numbers were locating in the range of 34-54 and double bond (DB) numbers were locating in the range of 0-6. In addition, the percentage of OPO, OPL, and OOO was relatively higher than others, accounted for more than 4% of total triglycerides in all the lactation areas and times, and the percentage of U2S and LLL triglycerides was also richest in mature milk. Furthermore, it was obvious that lactation regions had more significant effect on the triglycerides compared with lactation time and the triacylglycerols (TAGs) of human milk in Guangzhou were clearly different from that in other regions. Therefore, the results of this study will provide data reference for the design of infant formula suitable for Chinese babies.

Insight into the effects and biotechnological production of kestoses, the smallest fructooligosaccharides.

Kestoses, the smallest fructooligosaccharides, are trisaccharides composed of a fructose molecule and a sucrose molecule linked by either ß-(2,1) or ß-(2,6) linkage. 1-kestose, 6-kestose and neokestose are the three types of kestoses occurring in nature. As the main kind of fructooligosaccharide, kestoses share similar physiological effects with other fructooligosaccharides, and they have recently been determined to show more notable effects in promoting the growth of probiotics including Faecalibacterium prausnitzii and Bifidobacterium than those of other fructooligosaccharides. Kestoses exist in many plants, but the relatively low content and the isolation and purification are the main barriers limiting their industrial application. The production of kestoses by enzymatic biosynthesis and microbial fermentation has the potential to facilitate its production and industrial use. In this article, the recent advances in the research of kestoses were overviewed, including those studying their functions and production. Kestose-producing enzymes were introduced in detail, and microbial production and fermentation optimization techniques for enhancing the yield of kestoses were addressed. ß-Fructofuranosidase is the main one used to produce kestoses because of the extensive range of microbial sources. Therefore, the production of kestoses by microorganisms containing ß-fructofuranosidase has also been reviewed. However, few molecular modification studies have attempted to change the production profile of some enzymes and improve the yield of kestoses, which is a topic that should garner more attention. Additionally, the production of kestoses using food-grade microorganisms may be beneficial to their application in the food industry.