The monotreme pattern of egg-incubation followed by extended lactation represents the ancestral mammalian reproductive condition, suggesting that monotreme milk may include saccharides of an ancestral type. Saccharides were characterized from milk of the Tasmanian echidna Tachyglossus aculeatus setosus. Oligosaccharides in pooled milk from late lactation were purified by gel filtration and high-performance liquid chromatography using a porous graphitized carbon column and characterized by (1)H NMR spectroscopy; oligosaccharides in smaller samples from early and mid-lactation were separated by ultra-performance liquid chromatography and characterized by negative electrospray ionization mass spectrometry (ESI-MS) and tandem collision mass spectroscopy (MS/MS) product ion patterns. Eight saccharides were identified by (1)H NMR: lactose, 2'-fucosyllactose, difucosyllactose (DFL), B-tetrasaccharide, B-pentasaccharide, lacto-N-fucopentaose III (LNFP3), 4-O-acetyl-3'-sialyllactose [Neu4,5Ac(α2-3)Gal(β1-4)Glc] and 4-O-acetyl-3'-sialyl-3-fucosyllactose [Neu4,5Ac(α2-3)Gal(β1-4)[Fuc(α1-3)]Glc]. Six of these (all except DFL and LNFP3) were present in early and mid-lactation per ESI-MS, although some at trace levels. Four additional oligosaccharides examined by ESI-MS and MS/MS are proposed to be 3'-sialyllactose [Neu5Ac(α2-3)Gal(β1-4)Glc], di-O-acetyl-3'-sialyllactose [Neu4,5,UAc3(α2-3)Gal(β1-4)Glc where U = 7, 8 or 9], 4-O-acetyl-3'-sialyllactose sulfate [Neu4,5Ac(α2-3)Gal(β1-4)GlcS, where position of the sulfate (S) is unknown] and an unidentified 800 Da oligosaccharide containing a 4-O-acetyl-3'-sialyllactose core. 4-O-acetyl-3'-sialyllactose was the predominant saccharide at all lactation stages. 4-O-Acetylation is known to protect sialyllactose from bacterial sialidases and may be critical to prevent microbial degradation on the mammary areolae and/or in the hatchling digestive tract so that sialyllactose can be available for enterocyte uptake. The ability to defend against microbial invasion was probably of great functional importance in the early evolution of milk saccharides.
Previous structural characterizations of marsupial milk oligosaccharides had been performed in only two macropod species, the tammar wallaby and the red kangaroo. To clarify the homology and heterogeneity of milk oligosaccharides among marsupial species, which could provide information on their evolution, the oligosaccharides of the koala milk carbohydrate fraction were characterized in this study. Neutral and acidic oligosaccharides were separated from the carbohydrate fraction of milk of the koala, a non-macropod marsupial, and characterized by (1)H-nuclear magnetic resonance spectroscopy. The structures of the neutral saccharides were found to be Gal(β1-4)Glc (lactose), Gal(β1-3)Gal(β1-4)Glc (3'-galactosyllactose), Gal(β1-3)Gal(β1-3)Gal(β1-4)Glc (3',3″-digalactosyllactose), Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (lacto-N-novopentaose I) and Gal(β1-3){Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (fucosyl lacto-N-novopentaose I), while those of the acidic saccharides were Neu5Ac(α2-3)Gal(β1-4)Glc (3'-SL), Neu5Ac(α2-3)Gal(β1-3)Gal(β1-4)Gal (sialyl 3'-galactosyllactose), Neu5Ac(α2-3)Gal(β1-3)[Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose a), Gal(β1-3)[Neu5Ac(α2-6)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose b), Gal(β1-3)[Neu5Ac(α2-3)Gal(β1-4)GlcNAc(β1-6)]Gal(β1-4)Glc (sialyl lacto-N-novopentaose c), and Neu5Ac(α2-3)Gal(β1-3){Gal(β1-4)[Fuc(α1-3)]GlcNAc(β1-6)}Gal(β1-4)Glc (fucosyl sialyl lacto-N-novopentaose a). The neutral oligosaccharides, other than fucosyl lacto-N-novopentaose I, a novel hexasaccharide, had been found in milk of the tammar wallaby, a macropod marsupial, while the acidic oligosaccharides, other than fucosyl sialyl lacto-N-novopentaose a had been identified in milk carbohydrate of the red kangaroo. The presence of fucosyl oligosaccharides is a significant feature of koala milk, in which it differs from milk of the tammar wallaby and the red kangaroo.
The structures of milk oligosaccharides were characterized for four strepsirrhine primates to examine the extent to which they resemble milk oligosaccharides in other primates. Neutral and acidic oligosaccharides were isolated from milk of the greater galago (Galagidae: Otolemur crassicaudatus), aye-aye (Daubentoniidae: Daubentonia madagascariensis), Coquerel's sifaka (Indriidae: Propithecus coquereli) and mongoose lemur (Lemuridae: Eulemur mongoz), and their chemical structures were characterized by (1)H-NMR spectroscopy. The oligosaccharide patterns observed among strepsirrhines did not appear to correlate to phylogeny, sociality or pattern of infant care. Both type I and type II neutral oligosaccharides were found in the milk of the aye-aye, but type II predominate over type I. Only type II oligosaccharides were identified in other strepsirrhine milks. α3'-GL (isoglobotriose, Gal(α1-3)Gal(β1-4)Glc) was found in the milks of Coquerel's sifaka and mongoose lemur, which is the first report of this oligosaccharide in the milk of any primate species. 2'-FL (Fuc(α1-2)Gal(β1-4)Glc) was found in the milk of an aye-aye with an ill infant. Oligosaccharides containing the Lewis x epitope were found in aye-aye and mongoose lemur milk. Among acidic oligosaccharides, 3'-N-acetylneuraminyllactose (3'-SL-NAc, Neu5Ac(α2-3)Gal(β1-4)Glc) was found in all studied species, whereas 6'-N-acetylneuraminyllactose (6'-SL-NAc, Neu5Ac(α2-6)Gal(β1-4)Glc) was found in all species except greater galago. Greater galago milk also contained 3'-N-glycolylneuraminyllactose (3'-SL-NGc, Neu5Gc(α2-3)Gal(β1-4)Glc). The finding of a variety of neutral and acidic oligosaccharides in the milks of strepsirrhines, as previously reported for haplorhines, suggests that such constituents are ancient rather than derived features, and are as characteristic of primate lactation is the classic disaccharide, lactose.
The purpose of this study was to determine the quality (physical, chemical, microbiological characteristics), total phenolic content, and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) inhibition assay of probiotic yogurt supplemented with roselle flower extract (Hibiscus sabdariffa L) during cold storage. The experiment used treatment for types of yogurt as follows: cow's milk probiotic yogurt + roselle, goat's milk probiotic yogurt + roselle, cow's milk yogurt, and goat's milk yogurt. The yogurt was stored in cold storage and evaluated the quality and antioxidant activity variables on days 0, 3, 6, 9, 12, and 15 th. The results showed that there were interaction (P<0.05) between types of yogurt and storage time on pH value and total lactic acid bacteria (LAB), but no interaction effect on viscosity. The types of yogurt significantly affected (P<0.05) a w , total titrable acid (TTA), total phenolic content, and antioxidant activity. Cow's milk probiotic yogurt + roselle and goat's milk probiotic yogurt + roselle were the best yogurt that contributed to a good quality and high antioxidant activity up to 15 d at cold storage.
Dangke, a dairy product of cow or buffalo, is a traditional food of Enrekang, South Sulawesi Province. Addition of papain in dangke preparation is responsible for the formation of solid texture of dangke. This study was aimed to find optimum conditions (temperature and concentration of papain enzyme) and their effects on physical, chemical, microbiological, and hedonic qualities of dangke. This study consisted of two stages: preparation of papain and dangke production with heating temperatures (70, 80, and 90 °C) and papain treatments (0.2%, 0.3%, and 0.4%). The experiment was conducted in a completely randomized design with a 3 x 3 factorial arrangement with three replicates. The first factor was the processing temperature consisted of 3 levels i.e., 70, 80, and 90 °C. The second factor was the papain concentration consisted of 3 levels i.e., 0.2%, 0.3%, and 0.4%. The obtained data were evaluated using analysis of variance (ANOVA), followed by Duncan's Multiple Range Test to observe the significances among treatments. Papain and amino acids were characterized using descriptive methods and organoleptic study was performed by non-parametric test (Kruskal-Wallis). The highest protein concentration was found in commercial papain (Merck, 360.63 mg/100 g), while the protein content of papain used in this study was of 323.21 mg/100g. However, these enzymes had similar molecular weight of 19.17 kDa. The optimum condition of dangke preparation was found at heating temperature of 80 °C and 0.3% of papain concentration, resulting in the most desirable characteristics of dangke in terms of chemical, physical, and microbiological properties as well as hedonic evaluation. Keywords
The percentage of carbohydrate in the milk/colostrum of the mammalian is range from trace to over 10%, of which disaccharide lactose (Gal(β1-4)Glc) is usually constitutes the major part. Apart from the lactose (Gal(β1-4)Glc; Gal, D-galactose; Glc, D-glucose), the rest of carbohydrate components is composed of variety of sugars, commonly named as milk oligosaccharides. Human mature milk and colostrum contain 12 ~ 13 g/l and 22 ~ 24 g/l of oligosaccharides, respectively. In contrast, bovine colostrum contains more than 1 g/l oligosaccharides and this concentration rapidly decreases after 48 hr post partum. Most of human milk oligosaccharides (HMO) are resistant to digestion and absorption within the small intestine. Therefore they can reach the infant colon, where they can act as prebiotics that stimulate the growth of benefi cial microorganisms such as various species of Bifi dobacterium. They can also act as receptor analogues that inhibit the a achment of pathogenic microorganisms to the infant's colonic mucosa. A small part of the milk oligosaccharides is absorbed intact into the circulation and it has been hypothesized that these may act as immunomodulators. Generally, the bovine milk oligosaccharides are believed not to be absorbed by human adults or infants, thus making them available to be utilized as prebiotics or anti-infection materials. The colostrum of cows and other domestic farm animals is a potential source of free oligosaccharides, and oligosaccharides isolated from these natural sources can be utilized as functional foods or animal feedstuff s on the industrial scale.Key words: milk, oligosaccharides, prebiotic, Bifi dobacterium * Corresponding author: e-mail: urashima@obihiro.ac.jp oligosaccharides . In this review, we will introduce the structural feature of human milk oligosaccharides, on their fate within the gastrointestinal tract as well as on their possible biological functions as prebiotics, anti infection agents and immunomodulation factors. In addition, we will discuss the possibility of the commercial utilization of bovine milk oligosaccharides and those of other domestic farm animals. THE CHEMICAL STRUCTURES AND THEIR QUANTITATIVE ASPECTS OF HUMAN MILK OLIGOSACCHARIDESThe structures of at least 115 human milk oligosaccharides (HMOs) have been determined to date (Urashima et al., in press), while as many as 200 diff erent oligosaccharides have been separated and studied by microfl uidic high performance liquid chromatography
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