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Dietary exposure to heterocyclic amines in high-temperature cooked meat and fish in Malaysia

Jahurul MH, Jinap S, Ang SJ, Abdul-Hamid A, Hajeb P, Lioe HN, et al.

Food Addit Contam Part A Chem Anal Control Expo Risk Assess, 2010 Aug;27(8):1060-71.
PMID: 20589547 DOI: 10.1080/19440041003801190

The intake of heterocyclic amines is influenced by the amount and type of meat and fish ingested, frequency of consumption, cooking methods, cooking temperature, and duration of cooking. In this study, the dietary intake of heterocyclic amines in Malaysia and their main sources were investigated. Forty-two samples of meat and fish were analysed by high-performance liquid chromatography with photodiode array detector to determine the concentration of the six predominant heterocyclic amines, namely: 2-amino-3-methylimidazo[4,5-f] quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f] quinoline(MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f] quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimidazo[4,5-f] quinoxaline (4,8-DiMeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f] quinoxaline (7,8-DiMeIQx), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Dietary intake data were obtained using a food-frequency questionnaire when interviewing 600 Malaysian respondents. The level of total heterocyclic amines in food samples studies ranged from not detected to 38.7 ng g(-1). The average daily intake level of heterocyclic amine was 553.7 ng per capita day(-1). The intake of PhIP was the highest, followed by MeIQx and MeIQ. The results reveal that fried and grilled chicken were the major dietary source of heterocyclic amines in Malaysia. However, the heterocyclic amine intake by the Malaysian population was lower than those reported from other regions.
Fulltext Factors Contributing to Sharp Waste Disposal at Health Care Facility Among Diabetic Patients in North-East Peninsular Malaysia

Hasan UA, Mohd Hairon S, Yaacob NM, Daud A, Abdul Hamid A, Hassan N, et al.

Int J Environ Res Public Health, 2019 06 26;16(13).
PMID: 31247892 DOI: 10.3390/ijerph16132251

BACKGROUND: Type 2 diabetic patients are major users of medical sharps in the community. Proper sharp disposal practice among them, however, was reported to be low. The current study was aimed to determine the factors contributing to sharp waste disposal at a health care facility among Type 2 diabetic patients.
METHODS: In this cross-sectional study, Type 2 diabetic patients who were on insulin therapy attending health clinics were randomly selected and interviewed using a validated questionnaire. Binary logistic regression analysis was applied.
RESULTS: Out of 304 respondents, only 11.5% of them brought their used sharps to be disposed at health care facilities. Previous advice on sharp disposal from health care providers, knowledge score, and duration of diabetes were significant contributing factors for sharp waste disposal at health care facilities: (Adj. OR 6.31; 95% CI: 2.63, 15.12; p < 0.001), (Adj. OR 1.05; 95% CI: 1.03, 1.08; p < 0.001), and (Adj. OR 2.51; 95% CI: 1.06, 5.93; p = 0.036), respectively.
CONCLUSION: Continuous education and a locally adapted safe sharp disposal option must be available to increase awareness and facilitate diabetic patients adopting proper sharp disposal behavior.
Fulltext Effectiveness of Diabetes Community Sharp Disposal Education Module in Primary Care: An Experimental Study in North-East Peninsular Malaysia

Hasan UA, Mohd Hairon S, Yaacob NM, Daud A, Abdul Hamid A, Hassan N, et al.

Int J Environ Res Public Health, 2019 09 11;16(18).
PMID: 31514391 DOI: 10.3390/ijerph16183356

Background: Structured education is needed to cultivate safe sharp disposal behavior among diabetic patients. Thus, this study aimed to assess the effectiveness of the Diabetes Community Sharp Disposal Education Module in improving knowledge and sharp disposal practice among Malaysian Type 2 diabetic patients. Methods: This quasi-experimental study was conducted at primary health clinics in two districts in Kelantan, a state in the North-East Region of Peninsular Malaysia. A total of 132 Type 2 diabetic patients on insulin therapy were involved, with 68 participants in each control and intervention group. The health education intervention was based on the validated Diabetes Community Sharp Disposal Education Module. The knowledge and practices were measured using a validated questionnaire at baseline, one month, and three months after the intervention. Results: There was a significant increment in the mean knowledge score for intervention group; from baseline to one month follow up and from baseline to three months follow up [Greenhouse-Geisser; F(1.5, 199.7) = 62.38, p < 0.001; effect size (η2) = 0.318]. Intervention group had significantly higher mean knowledge score as compared to control group; at one month and three months follow up [F(1, 134) = 17.38, p < 0.001; effect size (η2) = 0.115]. There was a statistically significant increment in the proportion of participants in the intervention group who practiced the proper community sharp disposal method over time, X2(2) = 52.061, p < 0.001. Conclusions: The Diabetes Community Sharp Disposal Education Module was an effective health education tool to improve knowledge and encourage Malaysian diabetic patients to engage with proper sharp disposal practices.
Fulltext Evaluation of antioxidant compounds, antioxidant activities, and mineral composition of 13 collected purslane (Portulaca oleracea L.) accessions

Alam MA, Juraimi AS, Rafii MY, Abdul Hamid A, Aslani F, Hasan MM, et al.

Biomed Res Int, 2014;2014:296063.
PMID: 24579078 DOI: 10.1155/2014/296063

The methanolic extracts of 13 accessions of purslane were analyzed for their total phenol content (TPC), total flavonoid contents (TFC), and total carotenoid contents (TCC) and antioxidant activity of extracts was screened using FRAP assay and DPPH radical scavenging methods. The TPC, TFC, and TCC ranged from 0.96 ± 0.04 to 9.12 ± 0.29 mg GAE/g DW, 0.13 ± 0.04 to 1.44 ± 0.08 mg RE/g DW, and 0.52 ± 0.06 to 5.64 ± 0.09 mg (β-carotene equivalent) BCE/g DW, respectively. The DPPH scavenging (IC50) activity varied between 2.52 ± 0.03 mg/mL and 3.29 ± 0.01 mg/mL and FRAP ranged from 7.39 ± 0.08 to 104.2 ± 6.34 μmol TE/g DW. Among all the measured micro- and macrominerals K content was the highest followed by N, Na, Ca, Mg, P, Fe, Zn, and Mn. The overall findings proved that ornamental purslane was richer in antioxidant properties, whereas common purslane possesses more mineral contents than ornamental ones.
Fulltext Probiotics and Their Potential Preventive and Therapeutic Role for Cancer, High Serum Cholesterol, and Allergic and HIV Diseases

Nazir Y, Hussain SA, Abdul Hamid A, Song Y

Biomed Res Int, 2018;2018:3428437.
PMID: 30246019 DOI: 10.1155/2018/3428437

The potential health benefits of probiotics have long been elucidated since Metchnikoff and his coworkers postulated the association of probiotic consumption on human's health and longevity. Since then, many scientific findings and research have further established the correlation of probiotic and gut-associated diseases such as irritable bowel disease and chronic and antibiotic-associated diarrhea. However, the beneficial impact of probiotic is not limited to the gut-associated diseases alone, but also in different acute and chronic infectious diseases. This is due to the fact that probiotics are able to modify the intestinal microbial ecosystem, enhance the gut barrier function, provide competitive adherence to the mucosa and epithelium, produce antimicrobial substances, and modulate the immune activity by enhancing the innate and adaptive immune response. Nevertheless, the current literature with respect to the association of probiotic and cancer, high serum cholesterol, and allergic and HIV diseases are still scarce and controversial. Therefore, in the present work, we reviewed the potential preventive and therapeutic role of probiotics for cancer, high serum cholesterol, and allergic and HIV diseases as well as providing its possible mechanism of actions.
Inhibitory effect of morinda citrifolia L. On lipoprotein lipase activity

Pak-Dek MS, Abdul-Hamid A, Osman A, Soh CS

J Food Sci, 2008 Oct;73(8):C595-8.
PMID: 19019102 DOI: 10.1111/j.1750-3841.2008.00929.x

Efficacy of Morinda citrifolia L. leaf (MLE) and fruit extracts (MFE) in inhibiting lipoprotein lipase (LPL) was determined in vitro. The result of the study showed that the highest inhibition on the LPL activity was exhibited by MLE (66%+/- 2.1%), which is significantly higher than that demonstrated by MFE (54.5%+/- 2.5%), green tea extract (GTE) (54.5%+/- 2.6%), and catechin (43.6%+/- 6.1%). Percent of LPL inhibition increase with concentration of the extracts. Quantitative analysis of the extracts revealed the presence of high levels of (+)-catechin at 63.5 +/- 17 and 53.7 +/- 5.7 mg/g in MLE and MFE, respectively, although not as high as that found in GTE (530.6 +/- 42 mg/g). Appreciable amount of epicatechin was found in all extracts tested, while rutin was only found in MLE and MFE. The study suggested that both leaf and fruit of M. citrifolia may be used as antiobesity agents in body weight management.
Fulltext Executive function and attention span in euthymic patients with bipolar 1 disorder

Normala I, Abdul HA, Azlin B, Nik Ruzyanei NJ, Hazli Z, Shah SA

Med J Malaysia, 2010 Sep;65(3):199-203.
PMID: 21939168

This is a cross sectional comparison study to assess executive function and attention span in euthymic patients with bipolar 1 disorder. It compares the performance of these two cognitive domains in 40 patients with bipolar 1 disorder to that of 40 healthy normal subjects using Trail Making (TMT), Digit Span (Forward and Backward) and Verbal Fluency (VF) tests. The association between demographic, clinical characteristics and performance in all tests were examined. Patients with bipolar illness showed significant impairment with moderate to large effect sizes (VF = 0.67, TMT A = 0.52, TMT B = 0.81, Digit Forward = 0.97, Digit backward = 1.10) in all tasks of executive and attention functioning. These impairments are observed in the absence of active mood symptoms while duration and severity of illness are not found to have an effect on both cognitive domains. Medications received by patients with bipolar disorder have significant association with performance on executive tasks. The results of this study add on to the existing global evidence of cognitive impairment in bipolar illness despite its cross cultural differences. Its presence in the absence of mania, depression or mixed episode indicates that cognitive impairment is stable even after symptoms recovery.
Fulltext Thermal Annealing Effect on Structural, Morphological, and Sensor Performance of PANI-Ag-Fe Based Electrochemical E. coli Sensor for Environmental Monitoring

Mohammad Naim N, Abdullah H, Umar AA, Abdul Hamid A, Shaari S

ScientificWorldJournal, 2015;2015:696521.
PMID: 26078996 DOI: 10.1155/2015/696521

PANI-Ag-Fe nanocomposite thin films based electrochemical E. coli sensor was developed with thermal annealing. PANI-Ag-Fe nanocomposite thin films were prepared by oxidative polymerization of aniline and the reduction process of Ag-Fe bimetallic compound with the presence of nitric acid and PVA. The films were deposited on glass substrate using spin-coating technique before they were annealed at 300 °C. The films were characterized using XRD, UV-Vis spectroscopy, and FESEM to study the structural and morphological properties. The electrochemical sensor performance was conducted using I-V measurement electrochemical impedance spectroscopy (EIS). The sensitivity upon the presence of E. coli was measured in clean water and E. coli solution. From XRD analysis, the crystallite sizes were found to become larger for the samples after annealing. UV-Vis absorption bands for samples before and after annealing show maximum absorbance peaks at around 422 nm-424 nm and 426 nm-464 nm, respectively. FESEM images show the diameter size for nanospherical Ag-Fe alloy particles increases after annealing. The sensor performance of PANI-Ag-Fe nanocomposite thin films upon E. coli cells in liquid medium indicates the sensitivity increases after annealing.
Fulltext Relationship between antioxidant capacity and angiotensin-converting enzyme inhibitory activity of papain-generated protein hydrolysates and peptides from palm kernel cake proteins

Zarei, M., Abdul-Hamid, A., Muhialdin B. J., Saari, N.

International Food Research Journal, 2019;26(5):1641-1649.
MyJurnal

Palm kernel cake (PKC) proteins were hydrolysed in the presence of seven proteolytic enzymes.
The antioxidant and antihypertensive activities of each protein hydrolysate were evaluated.
Papain-generated protein hydrolysate showed the highest antioxidant effects measured by
radical scavenging activity (65%), iron (Fe)(II)-chelating activity (65%), and antihypertensive
activity (71%). The results revealed a strong correlation between the antioxidant activity of
the protein hydrolysates with angiotensin-converting enzyme (ACE) inhibitory activity (R2
= 0.69 - 0.98), except for trypsin-generated hydrolysate (R2 = 0.22). The most potent protein
hydrolysate was fractionated and profiled using reversed-phase high-performance liquid
chromatography and isoelectric focusing; peptides were subsequently isolated and identified
by tandem mass spectrometry. The individual peptides were evaluated for antihypertension
potential. A positive correlation was identified between radical scavenging activity and Fe(II)-
chelating activity together and ACE inhibitory activity with R2= 0.69 - 0.98. The findings indicate that there was a positive relationship between the antioxidant and antihypertensive activities of protein hydrolysates and bioactive peptides from PKC proteins.
Generation, Fractionation, and Characterization of Iron-Chelating Protein Hydrolysate from Palm Kernel Cake Proteins

Zarei M, Ghanbari R, Tajabadi N, Abdul-Hamid A, Bakar FA, Saari N

J Food Sci, 2016 Feb;81(2):C341-7.
PMID: 26720491 DOI: 10.1111/1750-3841.13200

Palm kernel cake protein was hydrolyzed with different proteases namely papain, bromelain, subtilisin, flavourzyme, trypsin, chymotrypsin, and pepsin to generate different protein hydrolysates. Peptide content and iron-chelating activity of each hydrolysate were evaluated using O-phthaldialdehyde-based spectrophotometric method and ferrozine-based colorimetric assay, respectively. The results revealed a positive correlation between peptide contents and iron-chelating activities of the protein hydrolysates. Protein hydrolysate generated by papain exhibited the highest peptide content of 10.5 mM and highest iron-chelating activity of 64.8% compared with the other hydrolysates. Profiling of the papain-generated hydrolysate by reverse phase high performance liquid chromatography fractionation indicated a direct association between peptide content and iron-chelating activity in most of the fractions. Further fractionation using isoelectric focusing also revealed that protein hydrolysate with basic and neutral isoelectric point (pI) had the highest iron-chelating activity, although a few fractions in the acidic range also exhibited good metal chelating potential. After identification and synthesis of papain-generated peptides, GGIF and YLLLK showed among the highest iron-chelating activities of 56% and 53%, whereas their IC50 were 1.4 and 0.2 μM, respectively.
Fulltext Angiotensin-I Converting Enzyme (ACE) Inhibitory and Anti-Oxidant Activities of Sea Cucumber (Actinopyga lecanora) Hydrolysates

Ghanbari R, Zarei M, Ebrahimpour A, Abdul-Hamid A, Ismail A, Saari N

Int J Mol Sci, 2015 Dec 04;16(12):28870-85.
PMID: 26690117 DOI: 10.3390/ijms161226140

In recent years, food protein-derived hydrolysates have received considerable attention because of their numerous health benefits. Amongst the hydrolysates, those with anti-hypertensive and anti-oxidative activities are receiving special attention as both activities can play significant roles in preventing cardiovascular diseases. The present study investigated the angiotensin-I converting enzyme (ACE) inhibitory and anti-oxidative activities of Actinopyga lecanora (A. lecanora) hydrolysates, which had been prepared by alcalase, papain, bromelain, flavourzyme, pepsin, and trypsin under their optimum conditions. The alcalase hydrolysate showed the highest ACE inhibitory activity (69.8%) after 8 h of hydrolysis while the highest anti-oxidative activities measured by 2,2-diphenyl 1-1-picrylhydrazyl radical scavenging (DPPH) (56.00%) and ferrous ion-chelating (FIC) (59.00%) methods were exhibited after 24 h and 8 h of hydrolysis, respectively. The ACE-inhibitory and anti-oxidative activities displayed dose-dependent trends, and increased with increasing protein hydrolysate concentrations. Moreover, strong positive correlations between angiotensin-I converting enzyme (ACE) inhibitory and anti-oxidative activities were also observed. This study indicates that A. lecanora hydrolysate can be exploited as a source of functional food owing to its anti-oxidant as well as anti-hypertension functions.
Fulltext The improvement of the endogenous antioxidant property of stone fish (Actinopyga lecanora) tissue using enzymatic proteolysis

Bordbar S, Ebrahimpour A, Abdul Hamid A, Abdul Manap MY, Anwar F, Saari N

Biomed Res Int, 2013;2013:849529.
PMID: 23586061 DOI: 10.1155/2013/849529

The stone fish (Actinopyga lecanora) ethanolic and methanolic tissue extracts were investigated for total phenolic contents (TPCs) as well as antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH(•)) radical scavenging activity and ferric reducing antioxidant power (FRAP) assays. Both extracts showed low amount of phenolics (20.33 to 17.03 mg of gallic acid equivalents/100 g dried sample) and moderate antioxidant activity (39% to 34% DPPH(•) radical scavenging activity and 23.95 to 22.30 mmol/100 mL FeSO4 FRAP value). Enzymatic proteolysis was carried out in order to improve the antioxidant activity using six commercially available proteases under their optimum conditions. The results revealed that the highest increase in antioxidant activity up to 85% was obtained for papain-generated proteolysate, followed by alcalase (77%), trypsin (75%), pepsin (68%), bromelain (68%), and flavourzyme (50%) as measured by DPPH(•) radical scavenging activity, whilst for the FRAP value, the highest increase in the antioxidant activity up to 39.2 mmol/100 mL FeSO4 was obtained for alcalase-generated proteolysate, followed by papain (29.5 mmol/100 mL FeSO4), trypsin (23.2 mmol/100 mL FeSO4), flavourzyme (24.7 mmol/100 mL FeSO4), bromelain (22.9 mmol/100 mL FeSO4), and pepsin (20.8 mmol/100 mL FeSO4). It is obvious that proteolysis of stone fish tissue by proteolytic enzymes can considerably enhance its antioxidant activity.
Fulltext Engineering the production of major catechins by Escherichia coli carrying metabolite genes of Camellia sinensis

Umar KM, Abdulkarim SM, Radu S, Abdul Hamid A, Saari N

ScientificWorldJournal, 2012;2012:529031.
PMID: 22645428 DOI: 10.1100/2012/529031

A mimicked biosynthetic pathway of catechin metabolite genes from C. sinensis, consisting of flavanone 3 hydroxylase (F3H), dihydroflavonol reductase (DFR), and leucoanthocyanidin reductase (LCR), was designed and arranged in two sets of constructs: (a) single promoter in front of F3H and ribosome-binding sequences both in front of DFR and LCR; (b) three different promoters with each in the front of the three genes and ribosome-binding sequences at appropriate positions. Recombinant E. coli BL (DE3) harbouring the constructs were cultivated for 65 h at 26 °C in M9 medium consisting of 40 g/L glucose, 1 mM IPTG, and 3 mM eriodictyol. Compounds produced were extracted in ethyl acetate in alkaline conditions after 1 h at room temperature and identified by HPLC. Two of the four major catechins, namely, (-)-epicatechin (0.01) and (-)-epicatechin gallate (0.36 mg/L), and two other types ((+)-catechin hydrate (0.13 mg/L) and (-)-catechin gallate (0.04 mg/L)) were successfully produced.
Fulltext Production of defatted palm kernel cake protein hydrolysate as a valuable source of natural antioxidants

Zarei M, Ebrahimpour A, Abdul-Hamid A, Anwar F, Saari N

Int J Mol Sci, 2012;13(7):8097-111.
PMID: 22942692 DOI: 10.3390/ijms13078097

The aim of this study was to produce a valuable protein hydrolysate from palm kernel cake (PKC) for the development of natural antioxidants. Extracted PKC protein was hydrolyzed using different proteases (alcalase, chymotrypsin, papain, pepsin, trypsin, flavourzyme, and bromelain). Subsequently, antioxidant activity and degree of hydrolysis (DH) of each hydrolysate were evaluated using DPPH• radical scavenging activity and O-phthaldialdehyde spectrophotometric assay, respectively. The results revealed a strong correlation between DH and radical scavenging activity of the hydrolysates, where among these, protein hydrolysates produced by papain after 38 h hydrolysis exhibited the highest DH (91 ± 0.1%) and DPPH• radical scavenging activity (73.5 ± 0.25%) compared to the other hydrolysates. In addition, fractionation of the most effective (potent) hydrolysate by reverse phase high performance liquid chromatography indicated a direct association between hydrophobicity and radical scavenging activity of the hydrolysates. Isoelectric focusing tests also revealed that protein hydrolysates with basic and neutral isoelectric point (pI) have the highest radical scavenging activity, although few fractions in the acidic range also exhibited good antioxidant potential.
Fulltext Actinopyga lecanora hydrolysates as natural antibacterial agents

Ghanbari R, Ebrahimpour A, Abdul-Hamid A, Ismail A, Saari N

Int J Mol Sci, 2012;13(12):16796-811.
PMID: 23222684 DOI: 10.3390/ijms131216796

Actinopyga lecanora, a type of sea cucumber commonly known as stone fish with relatively high protein content, was explored as raw material for bioactive peptides production. Six proteolytic enzymes, namely alcalase, papain, pepsin, trypsin, bromelain and flavourzyme were used to hydrolyze A. lecanora at different times and their respective degrees of hydrolysis (DH) were calculated. Subsequently, antibacterial activity of the A. lecanora hydrolysates, against some common pathogenic Gram positive bacteria (Bacillus subtilis and Staphylococcus aureus) and Gram negative bacteria (Escherichia coli, Pseudomonas aeruginosa, and Pseudomonas sp.) were evaluated. Papain hydrolysis showed the highest DH value (89.44%), followed by alcalase hydrolysis (83.35%). Bromelain hydrolysate after one and seven hours of hydrolysis exhibited the highest antibacterial activities against Pseudomonas sp., P. aeruginosa and E. coli at 51.85%, 30.07% and 30.45%, respectively compared to the other hydrolysates. Protein hydrolysate generated by papain after 8 h hydrolysis showed maximum antibacterial activity against S. aureus at 20.19%. The potent hydrolysates were further fractionated using RP-HPLC and antibacterial activity of the collected fractions from each hydrolysate were evaluated, wherein among them only three fractions from the bromelain hydrolysates exhibited inhibitory activities against Pseudomonas sp., P. aeruginosa and E. coli at 24%, 25.5% and 27.1%, respectively and one fraction of papain hydrolysate showed antibacterial activity of 33.1% against S. aureus. The evaluation of the relationship between DH and antibacterial activities of papain and bromelain hydrolysates revealed a meaningful correlation of four and six order functions.
Angiotensin-I Converting Enzyme (ACE) Inhibitory and Anti-Hypertensive Effect of Protein Hydrolysate from Actinopyga lecanora (Sea Cucumber) in Rats

Sadegh Vishkaei M, Ebrahimpour A, Abdul-Hamid A, Ismail A, Saari N

Mar Drugs, 2016 Sep 30;14(10).
PMID: 27706040

Food protein hydrolysates are known to exhibit angiotensin converting enzyme (ACE) inhibitory properties and can be used as a novel functional food for prevention of hypertension. This study evaluated the ACE inhibitory potentials of Actinopyga lecanora proteolysate (ALP) in vivo. The pre-fed rats with ALP at various doses (200, 400, 800 mg/kg body weight) exhibited a significant (p ≤ 0.05) suppression effect after inducing hypertension. To determine the optimum effective dose that will produce maximal reduction in blood pressure, ALP at three doses was fed to the rats after inducing hypertension. The results showed that the 800 mg/kg body weight dose significantly reduced blood pressure without noticeable negative physiological effect. In addition, there were no observable changes in the rats' heart rate after oral administration of the ALP. It was concluded that Actinopyga lecanora proteolysate could potentially be used for the development of functional foods and nutraceuticals for prevention and treatment of hypertension.
Fulltext Optimization of Bromelain-Aided Production of Angiotensin I-Converting Enzyme Inhibitory Hydrolysates from Stone Fish Using Response Surface Methodology

Muhammad Auwal S, Zarei M, Abdul-Hamid A, Saari N

Mar Drugs, 2017 Mar 31;15(4).
PMID: 28362352 DOI: 10.3390/md15040104

The stone fish is an under-utilized sea cucumber with many nutritional and ethno-medicinal values. This study aimed to establish the conditions for its optimum hydrolysis with bromelain to generate angiotensin I-converting enzyme (ACE)-inhibitory hydrolysates. Response surface methodology (RSM) based on a central composite design was used to model and optimize the degree of hydrolysis (DH) and ACE-inhibitory activity. Process conditions including pH (4-7), temperature (40-70 °C), enzyme/substrate (E/S) ratio (0.5%-2%) and time (30-360 min) were used. A pH of 7.0, temperature of 40 °C, E/S ratio of 2% and time of 240 min were determined using a response surface model as the optimum levels to obtain the maximum ACE-inhibitory activity of 84.26% at 44.59% degree of hydrolysis. Hence, RSM can serve as an effective approach in the design of experiments to improve the antihypertensive effect of stone fish hydrolysates, which can thus be used as a value-added ingredient for various applications in the functional foods industries.
High angiotensin-I converting enzyme (ACE) inhibitory activity of Alcalase-digested green soybean (Glycine max) hydrolysates

Hanafi MA, Hashim SN, Chay SY, Ebrahimpour A, Zarei M, Muhammad K, et al.

Food Res Int, 2018 04;106:589-597.
PMID: 29579964 DOI: 10.1016/j.foodres.2018.01.030

As a protein-rich, underutilized crop, green soybean could be exploited to produce hydrolysates containing angiotensin-I converting enzyme (ACE) inhibitory peptides. Defatted green soybean was hydrolyzed using four different food-grade proteases (Alcalase, Papain, Flavourzyme and Bromelain) and their ACE inhibitory activities were evaluated. The Alcalase-generated green soybean hydrolysate showed the highest ACE inhibitory activity (IC50: 0.14 mg/mL at 6 h hydrolysis time) followed by Papain (IC50: 0.20 mg/mL at 5 h hydrolysis time), Bromelain (IC50: 0.36 mg/mL at 6 h hydrolysis time) and Flavourzyme (IC50: 1.14 mg/mL at 6 h hydrolysis time) hydrolysates. The Alcalase-generated hydrolysate was profiled based on its hydrophobicity and isoelectric point using reversed phase high performance liquid chromatography (RP-HPLC) and isoelectric point focusing (IEF) fractionators. The Alcalase-generated green soybean hydrolysate comprising of peptides EAQRLLF, PSLRSYLAE, PDRSIHGRQLAE, FITAFR and RGQVLS, revealed the highest ACE inhibitory activity of 94.19%, 99.31%, 92.92%, 101.51% and 90.40%, respectively, while their IC50 values were 878 μM, 532 μM, 1552 μM, 1342 μM and 993 μM, respectively. It can be concluded that Alcalase-digested green soybean hydrolysates could be exploited as a source of peptides to be incorporated into functional foods with antihypertensive activity.
Fulltext Enzyme Hydrolysates from Stichopus horrens as a New Source for Angiotensin-Converting Enzyme Inhibitory Peptides

Forghani B, Ebrahimpour A, Bakar J, Abdul Hamid A, Hassan Z, Saari N

Evid Based Complement Alternat Med, 2012;2012:236384.
PMID: 22927875 DOI: 10.1155/2012/236384

Stichopus horrens flesh was explored as a potential source for generating peptides with angiotensin-converting enzyme (ACE) inhibitory capacity using 6 proteases, namely alcalase, flavourzyme, trypsin, papain, bromelain, and protamex. Degree of hydrolysis (DH) and peptide profiling (SDS-PAGE) of Stichopus horrens hydrolysates (SHHs) was also assessed. Alcalase hydrolysate showed the highest DH value (39.8%) followed by flavourzyme hydrolysate (32.7%). Overall, alcalase hydrolysate exhibited the highest ACE inhibitory activity (IC(50) value of 0.41 mg/mL) followed by flavourzyme hydrolysate (IC(50) value of 2.24 mg/mL), trypsin hydrolysate (IC(50) value of 2.28 mg/mL), papain hydrolysate (IC(50) value of 2.48 mg/mL), bromelain hydrolysate (IC(50) value of 4.21 mg/mL), and protamex hydrolysate (IC(50) value of 6.38 mg/mL). The SDS-PAGE results showed that alcalase hydrolysate represented a unique pattern compared to others, which yielded potent ACE inhibitory peptides with molecular weight distribution lower than 20 kDa. The evaluation of the relationship between DH and IC(50) values of alcalase and flavourzyme hydrolysates revealed that the trend between those parameters was related to the type of the protease used. We concluded that the tested SHHs would be used as a potential source of functional ACE inhibitory peptides for physiological benefits.
Fulltext Response Surface Optimisation for the Production of Antioxidant Hydrolysates from Stone Fish Protein Using Bromelain

Auwal SM, Zarei M, Abdul-Hamid A, Saari N

Evid Based Complement Alternat Med, 2017;2017:4765463.
PMID: 29234403 DOI: 10.1155/2017/4765463

Protein hydrolysates produced from different food sources exhibit therapeutic potential and can be used in the management of chronic diseases. This study was targeted to optimise the conditions for the hydrolysis of stone fish protein to produce antioxidant hydrolysates using central composite design (CCD) by response surface methodology (RSM). The stone fish protein was hydrolysed under the optimum predicted conditions defined by pH (6.5), temperature (54°C), E/S ratio (1.5%), and hydrolysis time (360 min). The hydrolysates were then evaluated for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•) scavenging activity and ferrous ion- (Fe2+-) chelating activity. Results validation showed no significant difference between the experimental values of DPPH• scavenging activity (48.94%) and Fe2+-chelating activity (25.12%) obtained at 54.62% degree of hydrolysis (DH) compared to their corresponding predicted values of 49.79% and 24.08% at 53.08% DH, respectively. The hydrolysates demonstrated non-Newtonian behavior (n < 1) with stronger shear-thinning effect and higher viscosities at increasing concentration. Thus, RSM can be considered as a promising strategy to optimise the production of stone fish protein hydrolysates containing antioxidant peptides. It is hoped that this finding will enhance the potential of stone fish protein hydrolysates (SHs) as therapeutic bioactive ingredient in functional foods development.

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