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Isolation and characterization of bioactive lactoferrin from camel milk by novel pH-dependent method for large scale production.

ABSTRACT: The present article exemplifies a novel method to isolate highly purified bioactive lactoferrin from camel milk. Cytotoxicity of lactoferrin against the Hela cells was used to evaluate its bioactivity. SDS-PAGE and LC-MS analysis was done for its identification and characterization. The purified camel milk lactoferrin was found to be 708 amino acids in length with a molecular weight of 77.3 kDa and a pI value of 8.24. This pH-dependent isolation procedure ensures the retention of bioactive lactoferrin from camel milk. The importance of the present work lies in its simplicity and scalability for manufacturing bioactive lactoferrin at an industrial level.

SUBMITTER: Mahala N

PROVIDER: S-EPMC9519473 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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Isolation and characterization of bioactive lactoferrin from camel milk by novel pH-dependent method for large scale production.

Mahala Neelam N Mittal Aastha A Lal Manohar M Dubey Uma S US

Biotechnology reports (Amsterdam, Netherlands) 20220921

The present article exemplifies a novel method to isolate highly purified bioactive lactoferrin from camel milk. Cytotoxicity of lactoferrin against the Hela cells was used to evaluate its bioactivity. SDS-PAGE and LC-MS analysis was done for its identification and characterization. The purified camel milk lactoferrin was found to be 708 amino acids in length with a molecular weight of 77.3 kDa and a pI value of 8.24. This pH-dependent isolation procedure ensures the retention of bioactive lacto ...[more]

PMID: 36188118

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Project description:As at 2020, Kenya was the best performing camel milk producer globally, with an annual production of 1.125 million tonnes. Despite the high production, about 50% of milk is wasted due to challenges affecting value addition to products such as yoghurt. The production of camel milk yoghurt faces multiple challenges, such as poor texture and weak structure, resulting in poor consumer acceptability. This study aimed to improve the physicochemical properties of camel milk yoghurt by adding different stabilising agents and calcium chloride. Yoghurt samples were processed using 3 L of camel milk, 6% sugar, 0.006% starter cultures, corn starch or modified starch and calcium chloride. The stabilisers were added at 2, 2.5, and 3% and Calcium Chloride at 0.075%. The milk was pasteurised at 90 °C for 30 min. Fermentation was performed for 6 h at 42 ±1 °C, and yoghurt was stored at 4 °C. The total titratable acidity, pH and viscosity were monitored hourly during fermentation and storage, while syneresis and water holding capacity were analysed at 1, 7, 14 and 21 days of refrigerated storage. The sensory evaluation was done using the 9-point hedonic scale to rate yoghurt samples' overall acceptability, colour, sweetness and thickness. The TTA of camel milk yoghurt increased with increasing fermentation time (0–6 h) and storage time from 1 to 21 days. The pH decreased with increasing fermentation time (0–6 h) and storage time from 1 to 21 days. The addition of stabilisers increased the viscosity of the yoghurt, with 3% corn starch exhibiting the highest viscosity throughout the fermentation and storage time. Corn starch had a higher effect on viscosity compared to modified starch. Calcium chloride further amplified the viscosity of the yoghurt. The addition of stabilisers reduced syneresis by over 44% compared to bovine yoghurt. In this study, the best results of viscosity, syneresis and sensory evaluation were observed when stabilising agents were added at the rate of 2.5% modified starch and 0.075% Calcium chloride. Corn starch, modified starch, camel milk yoghurt, physicochemical properties, rheological properties.

Project description:BackgroundMilk production traits are complex traits with vital economic importance in the camel industry. However, the genetic mechanisms regulating milk production traits in camels remain poorly understood. Therefore, we aimed to identify candidate genes and metabolic pathways that affect milk production traits in Bactrian camels.MethodsWe classified camels (fourth parity) as low- or high-yield, examined pregnant camels using B-mode ultrasonography, observed the microscopic changes in the mammary gland using hematoxylin and eosin (HE) staining, and used RNA sequencing to identify differentially expressed genes (DEGs) and pathways.ResultsThe average standard milk yield over the 300 days during parity was recorded as 470.18 ± 9.75 and 978.34 ± 3.80 kg in low- and high-performance camels, respectively. Nine female Junggar Bactrian camels were subjected to transcriptome sequencing, and 609 and 393 DEGs were identified in the low-yield vs. high-yield (WDL vs. WGH) and pregnancy versus colostrum period (RSQ vs. CRQ) comparison groups, respectively. The DEGs were compared with genes associated with milk production traits in the Animal Quantitative Trait Loci database and in Alashan Bactrian camels, and 65 and 46 overlapping candidate genes were obtained, respectively. Functional enrichment and protein-protein interaction network analyses of the DEGs and candidate genes were conducted. After comparing our results with those of other livestock studies, we identified 16 signaling pathways and 27 core candidate genes associated with maternal parturition, estrogen regulation, initiation of lactation, and milk production traits. The pathways suggest that emerged milk production involves the regulation of multiple complex metabolic and cellular developmental processes in camels. Finally, the RNA sequencing results were validated using quantitative real-time PCR; the 15 selected genes exhibited consistent expression changes.ConclusionsThis study identified DEGs and metabolic pathways affecting maternal parturition and milk production traits. The results provides a theoretical foundation for further research on the molecular mechanism of genes related to milk production traits in camels. Furthermore, these findings will help improve breeding strategies to achieve the desired milk yield in camels.

Dataset Information

Isolation and characterization of bioactive lactoferrin from camel milk by novel pH-dependent method for large scale production.

Publications

Isolation and characterization of bioactive lactoferrin from camel milk by novel pH-dependent method for large scale production.

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