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VERMA – Milk Protein Mining in Indian Goat Breeds

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VERMA – Milk Protein Mining in Indian Goat Breeds Milk Protein Mining in Indian Goat Breeds Mahima Verma1, Pramod K Rout1,* 1 Genetics and Breeding Division, ICAR-Central Institute for Research on Goats, Makhdoom, Farah, Mathura, Uttar Pradesh. India – 281122 2020 * corresponding author: [email protected], [email protected] Results Summary Milk protein association with CSN1S1 genotypes Protein% was significantly higher (P<0.05) in Himalayan local goat (4.31%), Ganjam (4.01%) and Gaddi (3.91%) as compared to other breeds. The goats with CSN1S1AB genotype had highest protein content (3.56%) followed by CSN1S1AA (3.50%), CSN1S1BB (3.26%), CSN1S1BF (3.03%) and CSN1S1AF (2.99%) genotypes. The CSN1S1AB genotype was producing 1.32 times more protein compared to CSN1S1FF genotype. Alleles CSN1S1A and CSN1S1B alleles were associated with higher protein content and occur in frequency 0.456, 0.502, respectively. Net Energy Milk yield at Protein % Fat % SNF % Lactose % (MJ/Kg) 90 days (Kg) Mean 3.386 ± 0.026 3.368 ± 0.189 9.007 ± 0.043 5.627 ± 0.035 3.058 ± 0.073 73.169± 0.2.012 CV 7.34 53.75 4.59 6.00 23.07 26.20 Introduction Range 2.46-4.51 1.01-8.9 7.72-10.51 4.14-7.28 2.062-5.360 314-1414 Heritability [Model: . Goat is a precious genetic resource for future agricultural productivity 0.441 ± 0.051 NE 0.294 ± 0.055 0.326 ± 0.053 NE NE Animal +pe] and climate change and Goat milk is used for family nutrition in disadvantage places. Proteomic characterization by 2DE and nLC-MS/MS . India is the highest producer of goat milk in the world and produces over The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the 5.6 million metric tons. PRIDE partner repository with the dataset identifier PXD013593. Goat milk is Identified 1067 peptides and 1307 proteins including casein and other low abundance proteins . Most HUMANIZED milk • Poteins were distributed in pI ranging 5 to 6 (24%), 74% of the proteins in the range of pI 5-9, 8% with pI <5 and Fulfills nutritional requirement of pregnant mothers, infants and 18% >9. older people in difficult conditions. • Major milk proteins caseins and whey were distributed in pI range of 4.28-9.7. Structures of lipid (fat globule) makes it easily digestible Gene Ontology analysis Lesser in lactose content Biological Process Terms: Metabolic process (18%), regulation of biological process (18.9%), defence response (8%), transport (10.7%), response to stimulus (16%) and cell organization and biogenesis (14.7%). Properties of hypo-allergenicity Cellular component Terms: Nucleus (18.1%), extracellular space (17.2%) cytoplasm (14.3%), chromosome (7.4%), Used for treatment of allergy, heat stress and respiratory diseases cytoskeleton (7.5%) and membrane (13.4%). Goat mammary gland is being used as a Bioreactor for production of Molecular function Terms: Protein binding (25.9%), catabolic activity (15.9%), structural molecule (14.9%), metal desired protein. ion binding (8.3%) and DNA binding (9.3%). Application in food and neutraceutical, cosmetic and fashion industry and pharmaceutical industry (Ayurveda drug formulation) Methodology Figure: Ontology The present study was carried out in 1450 milk samples of 15 Indian goat breeds/genotypes belonging to different agro-climatic zones. Analysis classification based on biological • Milk composition analysis was carried out by infrared spectroscopy process, molecular • Milk protein polymorphism was analysed by 14% SDS PAGE. function and cellular • CSN1S1 alleles were associated with milk composition traits component • 2DE was carried out for 32 identified protein variants • .raw files generated were analyzed using Proteome Discoverer (v2.2) against the in-house Uniprot proteome database. Biogrid interaction • Analysis was performed with goat, human, sheep and cow database with Annotations were available for 233 genes of the total identified proteome. Amyloid beta precursor protein had the 7056, 20117, 27666, and 23869 entries maximum count of 28, followed by heat shock protein family E (Hsp10) member 1 (count 12), keratin 40 (count 12). • Sequest search was carried out and both peptide spectrum match and Major interactions also involved deoxyuridine triphosphatase, tumor protein p53, abl interactor 2. protein false discovery rate were set to 0.01. Proteins associated in 144 KEGG pathways including information processing, metabolism and disease pathways such as Tuberculosis, Malaria, and Measles. Milk Systemic lupus erythematosus, alcoholism, viral carcinogenesis, metabolic pathways, biosynthesis of Goat yield antibiotics, tuberculosis, influenza A, legionellosis had the highest number of hits milk Composition Proteins involved in immune-regulation and mineral absorption like Antileukoproteinase, Neutrophil Protein% defensing, Thioredoxin, Clusterin, complement C3, Calcium-binding protein, Osteopontin and FABP were SNF% found to be present in goat milk. Fat% Lactose% Sixteen peptides identified in the database showed antimicrobial, ACE inhibitory, anti-inflammatory, anxiolytic, hypocholesterolemic, proliferating and opioid properties. Genotyping Name of genes identified in Tuberculosis pathway Digestion and Pre-fractionation • NF-κβ • Cathepsin D • Corosin -1 • Cathelicidin • IL-10 • Apaf -1 • C3b • HSP70 Tuberculosis Pathway and associated proteins Peptides and their functions The phosphosites observed for CSN2, CSN1S1, CSN1S2, CSN3 were 11P, 13P, 17P and 6P, respectively in 105 casein phosphopeptides. BLG and LALBA showed 19 and 4 phosphosites respectively PTM was observed in 45 low abundant non-casein milk proteins mainly associated with signal transduction, immune system, developmental biology and metabolism pathways. Conclusions: Applications of goat milk proteins and peptides in • Drug development • Nutraceuticals for human health application • Functional food development • Bioactive peptides and their functional role in health and nutrition ACKNOWLEDGEMENT: We express our gratitude to ICAR-NASF for funding this project. We also thank ICAR- CIRG for providing the required infrastructure. .
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