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  1. Fazilah, N. F., Zani, N. F. A., Wasoh, H., Ariff, A., Halim, M.
    MyJurnal
    Nowadays, functional food market is dominated by dairy-based probiotic products, mainly
    yogurt. The nutritional values of yogurt can be further enhanced by the inclusion of miracle
    fruit (Synsepalum dulcificum) and potential probiotic Lactococcus lactis Gh1. The present
    work investigated the anti-oxidative capacity and survivability of probiotic strains of six
    yogurts fortified with S. dulcificum pulp extract and encapsulated L. lactis Gh1 (in
    alginate-starch coating agent via extrusion technique). The flavonoid contents (TFC) were not
    significantly different between yogurts, whereas the phenolic contents (TPC) showed an
    increasing trend throughout the storage. Among the yogurts, the one supplemented with both
    S. dulcificum and encapsulated L. lactis Gh1 showed the highest TFC (1.18 µg QE/mL) and
    TPC (15.382 μg GAE/mL). The antioxidant assay (DPPH) showed a gradual increase on the
    first 7 d, but decreased afterward. In comparison, yogurts fortified with S. dulcificum demonstrated higher antioxidant activity (± 80% DPPH inhibition) than the plain yogurts (± 50%
    DPPH inhibition). The viability of starter cultures (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus) drastically increased during the first week (log 8 ~ 10
    CFU/mL) especially for yogurts containing free cell L. lactis, but subsequently decreased ( log
    6 ~ 8 CFU/mL). The viability of L. lactis Gh1 in yogurts maintained at high count (log 9.43
    and 9.04 CFU/mL) throughout 21 d when it was being encapsulated. In general, the fortification of S. dulcificum extract with microencapsulated L. lactis Gh1 had greatly enhanced the
    quality and potential benefits of the functional yogurts.
  2. Lim KT, Shukor MY, Wasoh H
    Biomed Res Int, 2014;2014:503784.
    PMID: 24696853 DOI: 10.1155/2014/503784
    Arsenic is a toxic metalloid which is widely distributed in nature. It is normally present as arsenate under oxic conditions while arsenite is predominant under reducing condition. The major discharges of arsenic in the environment are mainly due to natural sources such as aquifers and anthropogenic sources. It is known that arsenite salts are more toxic than arsenate as it binds with vicinal thiols in pyruvate dehydrogenase while arsenate inhibits the oxidative phosphorylation process. The common mechanisms for arsenic detoxification are uptaken by phosphate transporters, aquaglyceroporins, and active extrusion system and reduced by arsenate reductases via dissimilatory reduction mechanism. Some species of autotrophic and heterotrophic microorganisms use arsenic oxyanions for their regeneration of energy. Certain species of microorganisms are able to use arsenate as their nutrient in respiratory process. Detoxification operons are a common form of arsenic resistance in microorganisms. Hence, the use of bioremediation could be an effective and economic way to reduce this pollutant from the environment.
  3. Othman M, Ariff AB, Wasoh H, Kapri MR, Halim M
    AMB Express, 2017 Nov 27;7(1):215.
    PMID: 29181600 DOI: 10.1186/s13568-017-0519-6
    Lactic acid bacteria are industrially important microorganisms recognized for fermentative ability mostly in their probiotic benefits as well as lactic acid production for various applications. Fermentation conditions such as concentration of initial glucose in the culture, concentration of lactic acid accumulated in the culture, types of pH control strategy, types of aeration mode and different agitation speed had influenced the cultivation performance of batch fermentation of Pediococcus acidilactici. The maximum viable cell concentration obtained in constant fed-batch fermentation at a feeding rate of 0.015 L/h was 6.1 times higher with 1.6 times reduction in lactic acid accumulation compared to batch fermentation. Anion exchange resin, IRA 67 was found to have the highest selectivity towards lactic acid compared to other components studied. Fed-batch fermentation of P. acidilactici coupled with lactic acid removal system using IRA 67 resin showed 55.5 and 9.1 times of improvement in maximum viable cell concentration compared to fermentation without resin for batch and fed-batch mode respectively. The improvement of the P. acidilactici growth in the constant fed-batch fermentation indicated the use of minimal and simple process control equipment is an effective approach for reducing by-product inhibition. Further improvement in the cultivation performance of P. acidilactici in fed-bath fermentation with in situ addition of anion-exchange resin significantly helped to enhance the growth of P. acidilactici by reducing the inhibitory effect of lactic acid and thus increasing probiotic production.
  4. Mohd Zin NB, Mohamad Yusof B, Oslan SN, Wasoh H, Tan JS, Ariff AB, et al.
    AMB Express, 2017 Dec;7(1):131.
    PMID: 28651380 DOI: 10.1186/s13568-017-0433-y
    In recent years, many efforts have been directed to explore the methods to reduce the production costs of industrial lipase by improving the yield and the use of low-cost agricultural wastes. Coconut dregs, which is a lignocellulosic by-product from coconut oil and milk processing plants, is rich in cellulose (36%) and crude fat (9%). A newly isolated Bacillus stratosphericus has been demonstrated to perform cellulose hydrolysis on coconut dregs producing fermentable sugars. The highest extracellular lipase activity of 140 U/mL has been achieved in submerged fermentation with acid pre-treated coconut dregs. The lipase was found to be active over a wide range of temperatures and pHs. The activity of lipase can be generally increased by the presence of detergent ingredients such as Tween-80, cetyltrimethylammonium bromide, hydrogen peroxide and phosphate per sulphate. The great compatibility of lipase in commercial detergents has also underlined its potential as an additive ingredient in biodetergent formulations.
  5. Al Balawi AN, Yusof NA, Kamaruzaman S, Mohammad F, Wasoh H, Al-Lohedan HA
    Materials (Basel), 2019 Apr 11;12(7).
    PMID: 30978916 DOI: 10.3390/ma12071178
    The present study deals with the synthesis, characterization, and DNA extraction of poly(4,4'-cyclohexylidene bisphenol oxalate)/silica (Si) nanocomposites (NCs). The effects of varying the monomer/Si (3.7%, 7%, and 13%) ratio towards the size and morphology of the resulting NC and its DNA extraction capabilities have also been studied. For the NC synthesis, two different methods were followed, including the direct mixing of poly(4,4'-cyclohexylidene bisphenol oxalate) with fumed Si, and in situ polymerization of the 4,4'-cyclohexylidene bisphenol monomer in the presence of fumed silica (11 nm). The formed NCs were thoroughly investigated by using different techniques such as scanning electron microscopy (SEM), fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), powdered X-ray diffraction (XRD), and Brunauer-Emmett-Teller (BET) analysis where the results supported that there was the successful formation of poly(4,4'-cyclohexylidene bisphenol oxalate)/Si NC. Within the three different NC samples, the one with 13% Si was found to maintain a very high surface area of 12.237 m²/g, as compared to the other two samples consisting of 7% Si (3.362 m²/g) and 3.7% Si (1.788 m²/g). Further, the solid phase DNA extraction studies indicated that the efficiency is strongly influenced by the amount of polymer (0.2 g > 0.1 g > 0.02 g) and the type of binding buffer. Among the three binding buffers tested, the guanidine hydrochloride/EtOH buffer produced the most satisfactory results in terms of yield (1,348,000 ng) and extraction efficiency (3370 ng/mL) as compared to the other two buffers of NaCl (2 M) and phosphate buffered silane. Based on our results, it can be indicated that the developed poly(4,4'-cyclohexylidene bisphenol oxalate)/Si NC can serve as one of the suitable candidates for the extraction of DNA in high amounts as compared to other traditional solid phase approaches.
  6. Awaludin N, Abdullah J, Salam F, Ramachandran K, Yusof NA, Wasoh H
    Anal Biochem, 2020 12 01;610:113876.
    PMID: 32750357 DOI: 10.1016/j.ab.2020.113876
    The identification of rice bacterial leaf blight disease requires a simple, rapid, highly sensitive, and quantitative approach that can be applied as an early detection monitoring tool in rice health. This paper highlights the development of a turn-off fluorescence-based immunoassay for the early detection of Xanthomonas oryzae pv. oryzae (Xoo), a gram-negative bacterium that causes rice bacterial leaf blight disease. Antibodies against Xoo bacterial cells were produced as specific bio-recognition molecules and the conjugation of these antibodies with graphene quantum dots and gold nanoparticles was performed and characterized, respectively. The combination of both these bio-probes as a fluorescent donor and metal quencher led to changes in the fluorescence signal. The immunoreaction between AntiXoo-GQDs, Xoo cells, and AntiXoo-AuNPs in the immuno-aggregation complex led to the energy transfer in the turn-off fluorescence-based quenching system. The change in fluorescence intensity was proportional to the logarithm of Xoo cells in the range of 100-105 CFU mL-1. The limit of detection was achieved at 22 CFU mL-1 and the specificity test against other plant disease pathogens showed high specificity towards Xoo. The detection of Xoo in real plant samples was also performed in this study and demonstrated satisfactory results.
  7. Mohd Bakhori N, Yusof NA, Abdullah J, Wasoh H, Md Noor SS, Ahmad Raston NH, et al.
    Sensors (Basel), 2018 Jun 14;18(6).
    PMID: 29899214 DOI: 10.3390/s18061932
    In the present study, a beneficial approach for the ultrasensitive and affordable naked eye detection and diagnosis of tuberculosis (TB) by utilizing plasmonic enzyme-linked immunosorbent assay (ELISA) via antibody-antigen interaction was studied. Here, the biocatalytic cycle of the intracellular enzymes links to the formation and successive growth of the gold nanoparticles (GNPs) for ultrasensitive detection. The formation of different colored solutions by the plasmonic nanoparticles in the presence of enzyme labels links directly to the existence or non-existence of the TB analytes in the sample solutions. For disease detection, the adapted protocol is based mainly on the conventional ELISA procedure that involves catalase-labeled antibodies, i.e., the enzymes consume hydrogen peroxide and further produce GNPs with the addition of gold (III) chloride. The amount of hydrogen peroxide remaining in the solution determines whether the GNPs solution is to be formed in the color blue or the color red, as it serves as a confirmation for the naked eye detection of TB analytes. However, the conventional ELISA method only shows tonal colors that need a high concentration of analyte to achieve high confidence levels for naked eye detection. Also, in this research, we proposed the incorporation of protein biomarker, Mycobacterium tuberculosis ESAT-6-like protein esxB (CFP-10), as a means of TB detection using plasmonic ELISA. With the use of this technique, the CFP-10 detection limit can be lowered to 0.01 µg/mL by the naked eye. Further, our developed technique was successfully tested and confirmed with sputum samples from patients diagnosed with positive TB, thereby providing enough evidence for the utilization of our technique in the early diagnosis of TB disease.
  8. Ishak N, Lajis AFB, Mohamad R, Ariff AB, Mohamed MS, Halim M, et al.
    Molecules, 2018 Feb 24;23(2).
    PMID: 29495254 DOI: 10.3390/molecules23020501
    The synthesis of kojic acid derivative (KAD) from kojic and palmitic acid (C16:0) in the presence of immobilized lipase from Rhizomucor miehei (commercially known as Lipozyme RMIM), was studied using a shake flask system. Kojic acid is a polyfunctional heterocycles that acts as a source of nucleophile in this reaction allowing the formation of a lipophilic KAD. In this study, the source of biocatalyst, Lipozyme RMIM, was derived from the lipase of Rhizomucor miehei immobilized on weak anion exchange macro-porous Duolite ES 562 by the adsorption technique. The effects of solvents, enzyme loading, reaction temperature, and substrate molar ratio on the reaction rate were investigated. In one-factor-at-a-time (OFAT) experiments, a high reaction rate (30.6 × 10-3 M·min-1) of KAD synthesis was recorded using acetone, enzyme loading of 1.25% (w/v), reaction time of 12 h, temperature of 50 °C and substrate molar ratio of 5:1. Thereafter, a yield of KAD synthesis was optimized via the response surface methodology (RSM) whereby the optimized molar ratio (fatty acid: kojic acid), enzyme loading, reaction temperature and reaction time were 6.74, 1.97% (w/v), 45.9 °C, and 20 h respectively, giving a high yield of KAD (64.47%). This condition was reevaluated in a 0.5 L stirred tank reactor (STR) where the agitation effects of two impellers; Rushton turbine (RT) and pitch-blade turbine (PBT), were investigated. In the STR, a very high yield of KAD synthesis (84.12%) was achieved using RT at 250 rpm, which was higher than the shake flask, thus indicating better mixing quality in STR. In a rheological study, a pseudoplastic behavior of KAD mixture was proposed for potential application in lotion formulation.
  9. Mohd-Baseri N, Kheirel Anuar MS, Shamsuhazli NAS, Zulkifli MAF, Wasoh H, Yusof MT
    Int Microbiol, 2023 Jan;26(1):91-98.
    PMID: 36089618 DOI: 10.1007/s10123-022-00275-4
    Paddy is an important crop in Malaysia. There are various pathogens able to infect paddy causing a loss in yield's production. In this study, dual culture method, volatile organic compound (VOC) analysis, and non-volatile compound analysis were used to assess the ability of mushroom to control fungal rice pathogens including Curvularia lunata, Bipolaris panici-miliacei, and Nigrospora sp. Four mushroom isolates were further analysed for their antagonistic activity against rice pathogen. The highest percentage inhibition of radial growth (PIRG) was recorded between 45.55 and 73.68% observed in isolate 42b. The 4 isolates with the highest PIRG based on the dual culture analysis were then tested for their production of VOCs and non-volatile compound. Internal transcribed spacer (ITS) region analysis of the 4 mushroom isolates revealed their identity as Coprinellus disseminates (isolate 12b), Marasmiellus palmivorus (isolate 42b), Trametes maxima (isolate 56e), and Lentinus sajor-caju (isolate 60a). This study showed that mushroom isolates have the potential of antagonistic effect on various fungal rice pathogens tested by the production of secondary metabolites and mycoparasitic interaction.
  10. Fazilah NF, Hamidon NH, Ariff AB, Khayat ME, Wasoh H, Halim M
    Molecules, 2019 Apr 11;24(7).
    PMID: 30978923 DOI: 10.3390/molecules24071422
    There has been an explosion of probiotic incorporated based product. However, many reports indicated that most of the probiotics have failed to survive in high quantity, which has limited their effectiveness in most functional foods. Thus, to overcome this problem, microencapsulation is considered to be a promising process. In this study, Lactococcus lactis Gh1 was encapsulated via spray-drying with gum Arabic together with Synsepalum dulcificum or commonly known as miracle fruit. It was observed that after spray-drying, high viability (~10⁸ CFU/mL) powders containing L. lactis in combination with S. dulcificum were developed, which was then formulated into yogurt. The tolerance of encapsulated bacterial cells in simulated gastric juice at pH 1.5 was tested in an in-vitro model and the result showed that after 2 h, cell viability remained high at 1.11 × 10⁶ CFU/mL. Incubation of encapsulated cells in the presence of 0.6% (w/v) bile salts showed it was able to survive (~10⁴ CFU/mL) after 2 h. Microencapsulated L. lactis retained a higher viability, at ~10⁷ CFU/mL, when incorporated into yogurt compared to non-microencapsulated cells ~10⁵ CFU/mL. The fortification of microencapsulated and non-microencapsulated L. lactis in yogurts influenced the viable cell counts of yogurt starter cultures, Lactobacillus delbrueckii subs. bulgaricus and Streptococcus thermophilus.
  11. Mat Zaid MH, Che-Engku-Chik CEN, Yusof NA, Abdullah J, Othman SS, Issa R, et al.
    Molecules, 2020 Jul 24;25(15).
    PMID: 32722334 DOI: 10.3390/molecules25153373
    Death from tuberculosis has resulted in an increased need for early detection to prevent a tuberculosis (TB) epidemic, especially in closed and crowded populations. Herein, a sensitive electrochemical DNA biosensor based on functionalized iron oxide with mercaptopropionic acid (MPA-Fe3O4) nanoparticle and nanocellulose crystalline functionalized cetyl trimethyl ammonium bromide (NCC/CTAB) has been fabricated for the detection of Mycobacterium tuberculosis (MTB). In this study, a simple drop cast method was applied to deposit solution of MPA-Fe3O4/NCC/CTAB onto the surface of the screen-printed carbon electrode (SPCE). Then, a specific sequence of MTB DNA probe was immobilized onto a modified SPCE surface by using the 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling mechanism. For better signal amplification and electrochemical response, ruthenium bipyridyl Ru(bpy)32+ was assigned as labels of hybridization followed by the characteristic test using differential pulse voltammetry (DPV). The results of this biosensor enable the detection of target DNA until a concentration as low as 7.96 × 10-13 M with a wide detection range from 1.0 × 10-6 to 1.0 × 10-12 M. In addition, the developed biosensor has shown a differentiation between positive and negative MTB samples in real sampel analysis.
  12. Sharin SN, Sani MSA, Jaafar MA, Yuswan MH, Kassim NK, Manaf YN, et al.
    Food Chem, 2021 Jun 01;346:128654.
    PMID: 33461823 DOI: 10.1016/j.foodchem.2020.128654
    Identification of honey origin based on specific chemical markers is important for honey authentication. This study is aimed to differentiate Malaysian stingless bee honey from different entomological origins (Heterotrigona bakeri, Geniotrigona thoracica and Tetrigona binghami) based on physicochemical properties (pH, moisture content, ash, total soluble solid and electrical conductivity) and volatile compound profiles. The discrimination pattern of 75 honey samples was observed using Principal Component Analysis (PCA), Hierarchical Clustering Analysis (HCA), Partial Least Square-Discriminant Analysis (PLS-DA), and Support Vector Machine (SVM). The profiles of H. bakeri and G. thoracica honey were close to each other, but clearly separated from T. binghami honey, consistent with their phylogenetic relationship. T. binghami honey is marked by significantly higher electrical conductivity, moisture and ash content, and high abundance of 2,6,6-trimethyl-1-cyclohexene-1-carboxaldehyde, 2,6,6-trimethyl-1-cyclohexene-1-acetaldehyde and ethyl 2-(5-methyl-5-vinyltetrahydrofuran-2-yl)propan-2-yl carbonate. Copaene was proposed as chemical marker for G. thoracica honey. The potential of different parameters that aid in honey authentication was highlighted.
  13. Baskaran SM, Zakaria MR, Mukhlis Ahmad Sabri AS, Mohamed MS, Wasoh H, Toshinari M, et al.
    Environ Pollut, 2021 Feb 13;276:116742.
    PMID: 33621735 DOI: 10.1016/j.envpol.2021.116742
    Biodiesel side stream waste glycerol was identified as a cheap carbon source for rhamnolipids (RLs) production which at the same time could improve the management of waste. The present study aimed to produce RLs by using Pseudomonas aeruginosa RS6 utilizing waste glycerol as a substrate and to evaluate their physico-chemicals properties. Fermentation conditions such as temperature, initial medium pH, waste glycerol concentration, nitrogen sources and concentrations resulted in different compositions of the mono- and di-RLs produced. The maximum RLs production of 2.73 g/L was obtained when P. aeruginosa RS6 was grown in a basal salt medium supplemented with 1% waste glycerol and 0.2 M sodium nitrate at 35 °C and pH 6.5. At optimal fermentation conditions, the emulsification index (E24) values of cooking oil, diesel oil, benzene, olive oil, petroleum, and kerosene were all above E24=50%. The surface tension reduction obtained from 72.13 mN/m to 29.4-30.4 mN/m was better than the surface activity of some chemical-based surfactants. The RLs produced possessed antimicrobial activities against Gram-negative and Gram-positive bacteria with values ranging from 37% to 77% of growth inhibition when 1 mg/mL of RLs was used. Concentrations of RLs below 1500 μg/mL did not induce phytotoxicity effects on the tested seeds (Vigna radiata) compared to the chemical-based- surfactant, SDS. Furthermore, RLs tested on zebrafish (Danio rerio) embryos only exhibited low acute toxicity with an LC50 value of 72.97 μg/mL at 48 h of exposure suggesting a green and eco-biochemical worthy of future applications to replace chemical-based surfactants.
  14. Al Balawi AN, Yusof NA, Kamaruzaman S, Mohammad F, Wasoh H, Al Abbosh KF, et al.
    Biomed Res Int, 2019;2019:7064073.
    PMID: 30868072 DOI: 10.1155/2019/7064073
    The present study has synthesized poly(4,4'-cyclohexylidene bisphenol oxalate) by the condensation of oxalyl chloride with 4,4'-cyclohexylidene bisphenol, where its efficacy was tested for the solid-phase extraction of DNA. The synthesized polymer in the form of a white powder was characterized by FTIR, TGA-DTG, SEM, and BET analysis. The study utilized solid-phase application of the resulting polymer to extract DNA. The analysis of results provided the information that the extraction efficiency is a strong dependent of polymer amount and binding buffer type. Among the three types of buffers tested, the GuHCl buffer produced the most satisfactory results in terms of yield and efficiency of extraction. Moreover, the absorbance ratio of A260/A280 in all of the samples varied from 1.682 to 1.491, thereby confirming the capability of poly(4,4'-cyclohexylidene bisphenol oxalate) to elute pure DNA. The results demonstrated an increased DNA binding capacity with respect to increased percentage of the polymer. The study has concluded that poly(bisphenol Z oxalate) can be applied as one of the potential candidates for the high efficiency extraction of DNA by means of a simple, cost-effective, and environmentally friendly approach compared to the other traditional solid-phase methods.
  15. Gan Z, Roslan MAM, Abd Shukor MY, Halim M, Yasid NA, Abdullah J, et al.
    Biosensors (Basel), 2022 Oct 25;12(11).
    PMID: 36354431 DOI: 10.3390/bios12110922
    Aptamers are a group of synthetic single-stranded nucleic acids. They are generated from a random library of single-stranded DNA or RNA by a technology named systematic evolution of ligands by exponential enrichment (SELEX). SELEX is a repetitive process to select and identify suitable aptamers that show high affinity and specificity towards target cells. Great strides have been achieved in the design, construction, and use of aptamers up to this point. However, only a small number of aptamer-based applications have achieved widespread commercial and clinical acceptance. Additionally, finding more effective ways to acquire aptamers with high affinity remains a challenge. Therefore, it is crucial to thoroughly examine the existing dearth and advancement in aptamer-related technologies. This review focuses on aptamers that are generated by SELEX to detect pathogenic microorganisms and mammalian cells, as well as in cell-internalizing SELEX for diagnostic and therapeutic purposes. The development of novel aptamer-based biosensors using optical and electrical methods for microbial detection is reported. The applications and limitations of aptamers are also discussed.
  16. Ain Syaqirah Sapian N, Aidilfitri Mohamad Roslan M, Mohd Hashim A, Nasir Mohd Desa M, Halim M, Noorzianna Abdul Manaf Y, et al.
    Food Res Int, 2023 Feb;164:112332.
    PMID: 36737925 DOI: 10.1016/j.foodres.2022.112332
    Adulteration of lard with other fats and oils in food production affects many areas including economics, religion, and health. Previous studies discriminated lard based on major components of fats, i.e. triglycerides and fatty acids. This study aimed to differentiate lard and other animal fats (beef, chicken and mutton fat) based on n-alkane profiles established by gas chromatography-mass spectrometry (GC-MS). Principal Component Analysis (PCA) and Hierarchical Clustering Analysis (HCA) were able to initiate clustering of lard and other animal fats. Good result was obtained using Random Forest (RF) and Partial Least Squares-Discriminant Analysis (PLS-DA). Statistical models propose tetracosane (C24) as a potential n-alkane marker and it was found that C24 was the major alkane with composition of 15.72% (GC-MS) of total alkanes identified. Based on this finding, more interesting study may potentially be explored for the interest of various fats and oils consumers in vast applications especially using chemometrics analysis.
  17. Pannerchelvan S, Rios-Solis L, Faizal Wong FW, Zaidan UH, Wasoh H, Mohamed MS, et al.
    Food Funct, 2023 Mar 23.
    PMID: 36951915 DOI: 10.1039/d2fo03936b
    Gamma-aminobutyric acid (GABA) is a non-protein amino acid widely distributed in nature and extensively explored for its numerous physiological functions and effects on metabolic disorders. Lactic acid bacteria (LAB) are one of the most important GABA producers, vigorously pursued due to their high GABA content and generally regarded as safe (GRAS) status that allows for direct formulation in various GABA-enriched food products. To meet the strict requirements of the food and nutraceutical industries, the biosynthesis of GABA is typically preferred over the chemical synthesis route. The production of GABA varies among various strains of LAB and is affected by different fermentation conditions. Hence, optimizing the fermentation conditions to enhance the activity of the key enzyme glutamic acid decarboxylase is essential to maximize GABA production. This paper reviews the beneficial effects of GABA on human health and its applications in fermented food products. A particular emphasis is given to the biosynthetic approach for producing GABA by various LAB species via the microbial fermentation route. Efficient strategies for enhancing GABA production through optimization of the fermentation conditions, mode of fermentation, two-step fermentation, co-culturing approach, immobilization technique and genetic engineering are discussed in detail.
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