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Antibacterial Activity of Trypsin-Hydrolyzed Camel and Cow Whey and Their Fractions.

ABSTRACT: Antibacterial peptides were isolated and purified from whey proteins of camel milk (CaW) and cow milk (CoW) and their antimicrobial activities were studied. The whey proteins were hydrolyzed using trypsin, and the degree of hydrolysis was identified by gel electrophoresis. The whey hydrolysate (WH) was purified using ultrafiltration and Dextran gel chromatography to obtain small peptides with antibacterial activity. The effect of the antimicrobial peptides on the morphology of bacterial strains was investigated using transmission electron microscopy. Their amino acid composition and antimicrobial activities were then determined. Polypeptides CaWH-III (<3 kDa) and CoWH-III (<3 kDa) had the strongest antibacterial activity. Both Fr.A2 (CaWH-?'s fraction 2) and Fr.B1 (CoWH-?'s fraction 1) had antibacterial effects toward Escherichia coli and Staphylococcus aureus, with minimum antimicrobial mass concentrations of 65 mg/mL and 130 mg/mL for Fr.A2, and 130 mg/mL and 130 mg/mL for Fr.B1, respectively. The highly active antimicrobial peptides had high amounts of alkaline amino acids (28.13% in camel milk Fr.A2 and 25.07% in the cow milk Fr.B1) and hydrophobic amino acids. (51.29% in camel milk Fr.A2 and 57.69% in the cow milk Fr.B1). This results showed that hydrolysis of CaW and CoW using trypsin produced a variety of effective antimicrobial peptides against selected pathogens, and the antibacterial activity of camel milk whey was slightly higher than that of cow milk whey.

SUBMITTER: Wang R

PROVIDER: S-EPMC7070643 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Antibacterial Activity of Trypsin-Hydrolyzed Camel and Cow Whey and Their Fractions.

Wang Ruixue R Han Zhihao Z Ji Rimutu R Xiao Yuchen Y Si Rendalai R Guo Fucheng F He Jing J Hai Le L Ming Liang L Yi Li L

Animals : an open access journal from MDPI 20200220 2

Antibacterial peptides were isolated and purified from whey proteins of camel milk (CaW) and cow milk (CoW) and their antimicrobial activities were studied. The whey proteins were hydrolyzed using trypsin, and the degree of hydrolysis was identified by gel electrophoresis. The whey hydrolysate (WH) was purified using ultrafiltration and Dextran gel chromatography to obtain small peptides with antibacterial activity. The effect of the antimicrobial peptides on the morphology of bacterial strains ...[more]

PMID: 32093356

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Project description:Abstract Objectives The first year of life is a critical window for appropriate enteric microbiome development. Variations in infant diets have persistent effects on gut microbial ecology. Compared to intact protein formulas (IPF), partially hydrolyzed infant formulas (PHF) contain broken down milk proteins and may impact development of atopic conditions in high-risk infants. However, the effects of PHFs on the developing gut microbiome are not known. Methods This study was a secondary analysis of the GLOWING study (ClinicalTrials.gov NCT01131117). Infant stool samples were collected at 6 months of age and microbial composition was examined via 16S rRNA amplicon sequencing (V4 region) using standardized methods. Samples were grouped by primary feeding type: Exclusive breastfeeding (BF; n = 42), IPF (n = 17), or PHF (n = 12). We used linear models to examine the effects of feeding type adjusting for lifetime breastfeeding exposure at 6 mo. Group differences in α-diversity (richness and diversity) and relative abundances were assessed by ANOVA or t-test. Between-specimen diversity (β-diversity) was assessed using Bray − Curtis dissimilarities. Results Total breast milk exposure did not differ between PHF and IPF (mean 32.3% of feeds from human milk, p = 0.08). Analysis of α-diversity showed higher evenness (Shannon, Simpson) in IPF and higher richness (Chao1, Observed OTU) in PHF. However, these differences were no longer significant after adjusting for breastmilk exposure. β-diversity did not differ between groups. At the genus level compared to BF, infants fed IPF had higher abundance of Parabacteroides, while those fed PHF had higher Erwinia and Clostridium. Peptostreptococcaceae family was also increased in PHF fed infants. Compared to IPF, the PHF group had higher abundance of Clostridium genus, and Peptostreptococcaceae and Enterobacteriaceae families. Elevated Enterobacteriaceae and Clostridiales have previously been found in children with food allergies. Conclusions Our results suggest that partially hydrolyzed infant formulas may have modest impacts on the gut microbiome compared to intact protein formulas. Larger samples sizes are needed to fully characterize these differences and potential influence on long-term health. Funding Sources U.S. Department of Agriculture, NIDDK.

Project description:Human clinical trials have shown that a specific partially hydrolyzed 100% whey-based infant formula (pHF-W) reduces AD risk in the first yeast of life. Meta-analyses with a specific pHF-W (pHF-W1) confirm a protective effect while other meta-analyses pooling different pHF-W show conflicting results. Here we investigated the molecular composition and functional properties of the specific pHF-W1 as well as the stability of its manufacturing process over time. This specific pHF-W1 was compared with other pHF-Ws. We used size exclusion chromatography to characterize the peptide molecular weight (MW), a rat basophil degranulation assay to assess the relative level of beta-lactoglobulin (BLG) allergenicity and a preclinical model of oral tolerance induction to test prevention of allergic sensitization. To analyze the exact peptide sequences before and after an HLA binding assay, a mass cytometry approach was used. Peptide size allergenicity and oral tolerance induction were conserved across pHF-W1 batches of production and time. The median MW of the 37 samples of pHF-W1 tested was 800 ± 400 Da. Further oral tolerance induction was observed using 10 different batches of the pHF-W1 with a mean reduction of BLG-specific IgE levels of 0.76 log (95% CI = -0.95; -0.57). When comparing pHF-W1 with three other formulas (pHF-W2 3 and 4), peptide size was not necessarily associated with allergenicity reduction in vitro nor oral tolerance induction in vivo as measured by specific IgE level (p < 0.05 for pHF-W1 and 2 and p = 0.271 and p = 0.189 for pHF-W3 and 4 respectively). Peptide composition showed a limited overlap between the formulas tested ranging from 11.7% to 24.2%. Furthermore nine regions in the BLG sequence were identified as binding HLA-DR. In conclusion, not all pHF-Ws tested have the same peptide size distribution decreased allergenicity and ability to induce oral tolerance. Specific peptides are released during the different processes used by different infant formula producers.

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Antibacterial Activity of Trypsin-Hydrolyzed Camel and Cow Whey and Their Fractions.

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Antibacterial Activity of Trypsin-Hydrolyzed Camel and Cow Whey and Their Fractions.

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