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Characterization and identification of novel antidiabetic and anti-obesity peptides from camel milk protein hydrolysates

Mudgil P, Kamal H, Yuen GC, Maqsood S

Food Chem, 2018 Sep 01;259:46-54.
PMID: 29680061 DOI: 10.1016/j.foodchem.2018.03.082

In-vitro inhibitory properties of peptides released from camel milk proteins against dipeptidyl peptidase-IV (DPP-IV), porcine pancreatic α-amylase (PPA), and porcine pancreatic lipase (PPL) were studied. Results revealed that upon hydrolysis by different enzymes, camel milk proteins displayed dramatic increase in inhibition of DPP-IV and PPL, but slight improvement in PPA inhibition was noticed. Peptide sequencing revealed a total of 20 and 3 peptides for A9 and B9 hydrolysates respectively, obtained the score of 0.8 or more on peptide ranker and were categorized as potential DPP-IV inhibitory peptides. KDLWDDFKGL in A9 and MPSKPPLL in B9 were identified as most potent PPA inhibitory peptide. For PPL inhibition only 7 and 2 peptides qualified as PPL inhibitory peptides from hydrolysates A9 and B9, respectively. The present study report for the first time PPA and PPL inhibitory and only second for DPP-IV inhibitory potential of protein hydrolysates from camel milk.
Fulltext Psychosocial safety climate and self-efficacy: Moderating role of job-related expectations in Pakistani private-sector employees during the COVID-19 pandemic

Maqsood S, Sohail M, Naeem F, Nazri M, Fatima D

Front Psychol, 2022;13:1016050.
PMID: 36926534 DOI: 10.3389/fpsyg.2022.1016050

The labor force in Pakistan comprises 59.8 million individuals. The employees have faced major changes in work dynamics and psychosocial safety climate during the COVID-19 pandemic. The purpose of the current study is to find the relationship between psychosocial safety climate, self-efficacy, and job-related expectations. It explores the moderating role of job-related expectations on the relationship between psychosocial safety climate and self-efficacy. It was hypothesized that there is likely to be a significant relationship between psychosocial safety climate, self-efficacy, and job-related expectations, job-related expectations are likely to moderate the relationship between psychosocial safety climate and self-efficacy, and there are likely to be differences between married and unmarried employees; men and women; satisfied and unsatisfied employees with respect to psychosocial safety, self-efficacy, and job-related expectations. A correlational research design and a convenience sampling strategy were used. A total of 281 employees (M = 30.74 years, SD = 10.99) of the private-sector (including educational, industrial, and IT) organizations during the COVID-19 pandemic were part of the study. Results show that psychosocial safety climate had a positive significant relationship with job-related expectations and self-efficacy. Job expectations also significantly correlated with self-efficacy. There were significant differences in measures of study variables with respect to gender, marital status, and employee satisfaction. This research has implications for administration, managers, policymakers, and organizational psychologists.
Identification and molecular docking study of novel cholesterol esterase inhibitory peptides from camel milk proteins

Mudgil P, Baby B, Ngoh YY, Vijayan R, Gan CY, Maqsood S

J Dairy Sci, 2019 Dec;102(12):10748-10759.
PMID: 31548068 DOI: 10.3168/jds.2019-16520

Novel bioactive peptides from camel milk protein hydrolysates (CMPH) were identified and tested for inhibition of cholesterol esterase (CEase), and their possible binding mechanisms were elucidated by molecular docking. Papain-generated CMPH showed the highest degree of hydrolysis. All CMPH produced upon enzymatic degradation demonstrated a dramatic enhancement of CEase inhibition compared with intact camel milk proteins, with papain-generated hydrolysate P9 displaying the highest inhibition. Peptide identification and their modeling through PepSite 2 revealed that among 20 potential bioactive peptides in alcalase-generated hydrolysate A9, only 3 peptides, with sequences KFQWGY, SQDWSFY, and YWYPPQ, showed the highest binding toward CEase catalytic sites. Among 43 peptides in 9-h papain-generated hydrolysate P9, 4 peptides were found to be potent CEase inhibitors. Molecular docking revealed that WPMLQPKVM, CLSPLQMR, MYQQWKFL, and CLSPLQFR from P9 hydrolysates were able to bind to the active site of CEase with good docking scores and molecular mechanics-generalized born surface area binding energies. Overall, this is the first study reporting CEase inhibitory potential of peptides generated from milk proteins.
Identification and characterization of novel α-amylase and α-glucosidase inhibitory peptides from camel whey proteins

Baba WN, Mudgil P, Kamal H, Kilari BP, Gan CY, Maqsood S

J Dairy Sci, 2021 Feb;104(2):1364-1377.
PMID: 33309363 DOI: 10.3168/jds.2020-19271

This study explores the inhibitory properties of camel whey protein hydrolysates (CWPH) toward α-amylase (AAM) and α-glucosidase (AG). A general full factorial design (3 × 3) was applied to study the effect of temperature (30, 37, and 45°C), time (120, 240, and 360 min), and enzyme (pepsin) concentration (E%; 0.5, 1, and 2%). The results showed that maximum degree of hydrolysis was obtained when hydrolysis was carried out at higher temperature (45°C; P < 0.05), compared with lower temperatures of 30 and 37°C. Electrophoretic pattern displays degradation of all protein bands upon hydrolysis by pepsin at various hydrolysis conditions applied. All the 27 CWPH generated showed significant AAM and AG inhibitory potential as indicated by their lower IC50 values (mg/mL) compared with intact whey proteins. In total 196 peptides were identified from selected hydrolysates and 15 potential peptides (PepSite score > 0.8; http://pepsite2.russelllab.org/) were explored via in silico approach. Novel peptides PAGNFLMNGLMHR, PAVACCLPPLPCHM, MLPLMLPFTMGY, and PAGNFLPPVAAAPVM were identified as potential inhibitors for both AAM and AG due to their high number of binding sites and highest binding probability toward the target enzymes. CCGM and MFE, as well as FCCLGPVPP were identified as AG and AAM inhibitory peptides, respectively. This is the first study that reports novel AG and AAM inhibitory peptides from camel whey proteins. The future direction for this research involves synthesis of these potential AG and AAM inhibitory peptides in a pure form and investigate their antidiabetic properties in the in vitro, as well as in vivo models. Thus, CWPH can be considered for potential applications in glycaemic regulation.
Molecular basis of the anti-diabetic properties of camel milk through profiling of its bioactive peptides on dipeptidyl peptidase IV (DPP-IV) and insulin receptor activity

Ashraf A, Mudgil P, Palakkott A, Iratni R, Gan CY, Maqsood S, et al.

J Dairy Sci, 2021 Jan;104(1):61-77.
PMID: 33162074 DOI: 10.3168/jds.2020-18627

The molecular basis of the anti-diabetic properties of camel milk reported in many studies and the exact active agent are still elusive. Recent studies have reported effects of camel whey proteins (CWP) and their hydrolysates (CWPH) on the activities of dipeptidyl peptidase IV (DPP-IV) and the human insulin receptor (hIR). In this study, CWPH were generated, screened for DPP-IV binding in silico and inhibitory activity in vitro, and processed for peptide identification. Furthermore, pharmacological action of intact CWP and their selected hydrolysates on hIR activity and signaling and on glucose uptake were investigated in cell lines. Results showed inhibition of DPP-IV by CWP and CWPH and their positive action on hIR activation and glucose uptake. Interestingly, the combination of CWP or CWPH with insulin revealed a positive allosteric modulation of hIR that was drastically reduced by the competitive hIR antagonist. Our data reveal for the first time the profiling and pharmacological actions of CWP and their derived peptides fractions on hIR and their pathways involved in glucose homeostasis. This sheds more light on the anti-diabetic properties of camel milk by providing the molecular basis for the potential use of camel milk in the management of diabetes.
New insights into the cholesterol esterase- and lipase-inhibiting potential of bioactive peptides from camel whey hydrolysates: Identification, characterization, and molecular interaction

Baba WN, Mudgil P, Baby B, Vijayan R, Gan CY, Maqsood S

J Dairy Sci, 2021 Jul;104(7):7393-7405.
PMID: 33934858 DOI: 10.3168/jds.2020-19868

Novel antihypercholesterolemic bioactive peptides (BAP) from peptic camel whey protein hydrolysates (CWPH) were generated at different time, temperature, and enzyme concentration (%). Hydrolysates showed higher pancreatic lipase- (PL; except 3 CWPH) and cholesterol esterase (CE)-inhibiting potential, as depicted by lower half-maximal inhibitory concentration values (IC50 values) compared with nonhydrolyzed camel whey proteins (CWP). Peptide sequencing and in silico data depicted that most BAP from CWPH could bind active site of PL, whereas as only 3 peptides could bind the active site of CE. Based on higher number of reactive residues in the BAP and greater number of substrate binding sites, FCCLGPVPP was identified as a potential CE-inhibitory peptide, and PAGNFLPPVAAAPVM, MLPLMLPFTMGY, and LRFPL were identified as PL inhibitors. Molecular docking of selected peptides showed hydrophilic and hydrophobic interactions between peptides and target enzymes. Thus, peptides derived from CWPH warrant further investigation as potential candidates for adjunct therapy for hypercholesterolemia.
Simulated gastrointestinal digestion of camel and bovine casein hydrolysates: Identification and characterization of novel anti-diabetic bioactive peptides

Mudgil P, Kamal H, Priya Kilari B, Mohd Salim MAS, Gan CY, Maqsood S

Food Chem, 2021 Aug 15;353:129374.
PMID: 33740505 DOI: 10.1016/j.foodchem.2021.129374

Camel milk proteins are an important substrate for bioactive peptides generation. This study investigates in-vitro antidiabetic effect (via inhibition of α-amylase (AA), α-glucosidase (AG) and dipeptidyl peptidase IV (DPP-IV)) of bovine (BC) and camel casein (CC) hydrolysates. Further, effect of simulated gastrointestinal digestion (SGID) on inhibitory potential of generated hydrolysates was also explored. Both BC and CC hydrolysates displayed potent inhibitory properties against AA (IC50 value- 0.58 & 0.59 mg/mL), AG (IC50 value- 1.04 & 0.59 mg/mL) and DPP-IV (IC50 value- 0.62 & 0.66 mg/mL), respectively. Among different peptides identified in BC and CC hydrolysates, it was observed that FLWPEYGAL was predicted to be most potent inhibitory peptide against AA. While LPTGWLM, MFE and GPAHCLL as most active inhibitor of AG and HLPGRG, QNVLPLH and PLMLP were predicted to be active against DPP-IV. Overall, BC and CC hydrolysates can be proposed to be used in different food formulations as functional antidiabetic agents.
Fulltext Camel whey protein hydrolysates induced G2/M cellcycle arrest in human colorectal carcinoma

Murali C, Mudgil P, Gan CY, Tarazi H, El-Awady R, Abdalla Y, et al.

Sci Rep, 2021 03 29;11(1):7062.
PMID: 33782460 DOI: 10.1038/s41598-021-86391-z

Camel milk has been gaining immmense importance due to high nutritious value and medicinal properties. Peptides from milk proteins is gaining popularity in various therapeutics including human cancer. The study was aimed to investigate the anti-cancerous and anti-inflammatory properties of camel whey protein hydrolysates (CWPHs). CWPHs were generated at three temperatures (30 ℃, 37 ℃, and 45 ℃), two hydrolysis timepoints (120 and 360 min) and with three different enzyme concentrations (0.5, 1 and 2 %). CWPHs demonstrated an increase in anti-inflammatory effect between 732.50 (P-6.1) and 3779.16 (P-2.1) µg Dicolfenac Sodium Equivalent (DSE)/mg protein. CWPHs (P-4.3 & 5.2) inhibited growth of human colon carcinoma cells (HCT116) with an IC50 value of 231 and 221 μg/ml, respectively. P-4.3 induced G2/M cell cycle arrest and modulated the expression of Cdk1, p-Cdk1, Cyclin B1, p-histone H3, p21 and p53. Docking of two peptides (AHLEQVLLR and ALPNIDPPTVER) from CWPHs (P-4.3) identified Polo like kinase 1 as a potential target, which strongly supports our in vitro data and provides an encouraging insight into developing a novel peptide-based anticancer formulation. These results suggest that the active component, CWPHs (P-4.3), can be further studied and modeled to form a small molecule anti-cancerous therapy.
Camel Milk-Derived Bioactive Peptides and Diabetes: Molecular View and Perspectives

Ayoub MA, Yap PG, Mudgil P, Khan FB, Anwar I, Muhammad K, et al.

J Dairy Sci, 2023 Sep 12.
PMID: 37709024 DOI: 10.3168/jds.2023-23733

In dairy science, camel milk (CM) constitutes a center of interest for scientists due to its known beneficial impact on diabetes as demonstrated in many in vitro, in vivo, and clinical studies and trials. Overall, CM had positive effects on various parameters related to glucose transport and metabolism as well as the structural and functional properties of the pancreatic β-cells and insulin secretion. Thus, CM consumption may help manage diabetes, however, such a recommendation will become rationale and clinically conceivable only if the exact molecular mechanisms and pathways involved at the cellular levels are well understood. Moreover, the application of CM as an alternative antidiabetic tool may first require the identification of the exact bioactive molecule(s) behind such antidiabetic properties. In this review, we describe the advances in our knowledge of the molecular mechanisms reported to be involved in the beneficial effects of CM in managing diabetes using different in vitro and in vivo models. This mainly includes the effects of CM on the different molecular pathways controlling (i) insulin receptor signaling and glucose uptake, (ii) the pancreatic β-cell structure and function, and (iii) the activity of key metabolic enzymes in glucose metabolism. Moreover, we described the current status of the identification of CM-derived bioactive peptides and their structure-activity relationship study and characterization in the context of molecular markers related to diabetes. Such an overview will not only enrich our scientific knowledge of the plausible mode of action of CM in diabetes but should ultimately rationalize the claim of the potential application of CM against diabetes. This will pave the way toward new directions and ideas for developing a new generation of antidiabetic products taking benefits from the chemical composition of CM.
Fulltext Exploring the impact of various cooking techniques on the physicochemical and quality characteristics of camel meat product

Bahwan M, Baba WN, Adiamo O, Hassan HM, Roobab U, Abayomi OO, et al.

Anim Biosci, 2023 Nov;36(11):1747-1756.
PMID: 37402461 DOI: 10.5713/ab.22.0238

OBJECTIVE: The objective of this study was to evaluate the effects of four different cooking techniques viz: boiling, grilling, microwave, and frying; on the physicochemical characteristics of camel meat.
METHODS: Protein composition and their degradation as well as biochemical and textural changes of camel meat as influenced by cooking methods were investigated.
RESULTS: The highest cooking loss (52.61%) was reported in microwaved samples while grilled samples showed the lowest cooking loss (44.98%). The microwaved samples showed the highest levels of lipid oxidation as measured by thiobarbituric acid reactive substances, while boiled samples showed the lowest levels (4.5 mg/kg). Protein solubility, total collagen, and soluble collagen content were highest in boiled samples. Boiled camel meat had lower hardness values compared to the other treated samples. Consequently, boiling was the more suitable cooking technique for producing camel meat with a reduced hardness value and lower lipid oxidation level.
CONCLUSION: The camel meat industry and camel meat consumer can benefit from this research by improving their commercial viability and making consumers aware about the effects of cooking procedures on the quality of camel meat. The results of this study will be of significance to researchers and readers who are working on the processing and quality of camel meat.
A comparative investigation into novel cholesterol esterase and pancreatic lipase inhibitory peptides from cow and camel casein hydrolysates generated upon enzymatic hydrolysis and in-vitro digestion

Mudgil P, Baba WN, Kamal H, FitzGerald RJ, Hassan HM, Ayoub MA, et al.

Food Chem, 2022 Jan 15;367:130661.
PMID: 34348197 DOI: 10.1016/j.foodchem.2021.130661

Cow (CwC) and camel casein (CaC) hydrolysates were generated using Alcalase™ (CwCA and CaCA) and Pronase-E (CwCP and CaCP) each for 3 and 6 h, and investigated for their potential to inhibit key lipid digesting enzymes i.e., pancreatic lipase (PL) and cholesteryl esterase (CE). Results revealed stronger PL and CE inhibition by CaC hydrolysates compared to CwC. Potent hydrolysates (CwCP-3 h and CaCA-6 h) upon simulated gastrointestinal digestion (SGID) showed significant improvement in inhibition of both PL and CE. However, both the SGID hydrolysates showed similar extent of PL and CE inhibition and were further sequenced for peptide identification. Peptides MMML, FDML, HLPGRG from CwC and AAGF, MSNYF, FLWPEYGAL from CaC hydrolysates were predicted to be most active PL inhibitory peptides. Peptide LP found in both CwC and CaC hydrolysates was predicted as active CE inhibitor. Thus, CwC and CaC could be potential source of peptides with promising CE and PL inhibitory properties.
Fulltext Molecular docking studies on α-amylase inhibitory peptides from milk of different farm animals

Mudgil P, Al Dhaheri MKO, Alsubousi MSM, Khan H, Redha AA, Yap PG, et al.

J Dairy Sci, 2024 May;107(5):2633-2652.
PMID: 38101739 DOI: 10.3168/jds.2023-24118

Milk-derived peptides have emerged as a popular mean to manage various lifestyle disorders such as diabetes. Fermentation is being explored as one of the faster and efficient way of producing peptides with antidiabetic potential. Therefore, in this study, an attempt was made to comparatively investigate the pancreatic α-amylase (PAA) inhibitory properties of peptides derived from milk of different farm animals through probiotic fermentation. Peptide's identification was carried out using liquid chromatography-quadrupole time-of-flight mass spectrometry and inhibition mechanisms were characterized by molecular docking. Results obtained showed a PAA-IC50 value (the amount of protein equivalent needed to inhibit 50% of enzymes) between 2.39 and 36.1 µg protein equivalent for different fermented samples. Overall, Pediococcus pentosaceus MF000957-derived fermented milk from all animals indicated higher PAA inhibition than other probiotic derived fermented milk (PAA-IC50 values of 6.01, 3.53, 15.6, and 10.8 µg protein equivalent for bovine, camel, goat, and sheep fermented milk). Further, molecular docking analysis indicated that camel milk-derived peptide IMEQQQTEDEQQDK and goat milk-derived peptide DQHQKAMKPWTQPK were the most potent PAA inhibitory peptides. Overall, the study concluded that fermentation derived peptides may prove useful in for managing diabetes via inhibition of carbohydrate digesting enzyme PAA.

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