Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF) of palm kernel cake (PKC). The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30°C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w) on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.
The blending effects of mango seed fat (MSF), extracted using supercritical fluid, and palm stearin (PS) to formulate hard cocoa butter replacers (CBRs), were investigated. The triglycerides (TG), thermal properties and solid fat content (SFC) of the formulated blends were determined using different chromatographic and thermal techniques. All the blends had three main TGs; namely, 1,3-dipalmitoyl-2-oleoylglycerol (POP) (8.6-17.7%), 1-palmitoyl-2-oleoyl-3-stearoyl-glycerol (POS) (12.6-19.6%), and 1,3-distearoyl-2-oleoyl-glycerol (SOS) (37.2-31.4%), with SOS being the major component. The melting peak temperatures gradually increased and shifted towards higher temperatures with PS. The crystallization onset temperatures increased, while the offset decreased with PS. The SFC did not drop to 0% at 37.5°C, which was shifted to 0% at and above 40°C for some blends. The studies revealed that CBRs could be prepared by blending MSF and PS, and they could be utilised by chocolate manufacturers in tropical countries.
This research was conducted in order to investigate rumen degradability of some factors includes; Dray Matter (DM), Organic Matter (OM), Crude Protein (CP), Acid Detergent Fiber (ADF) in three different plant protein supplements includes; soybean, canola and cottonseed meal. The experiment was carried out using in three castrated and fistulated male Zel sheep. Each feedstuff was weighed into duplicate nylon bags and incubated in each of the three rumen fistulated sheep for 0, 2, 4, 8, 16, 24 and 48 h. Results revealed that effective degradability of DM of soybean, canola and cottonseed meal were 55.8, 73.8 and 48.5%, respectively. Effective degradabilities of the CP in feedstuffs were 55.8, 62 and 48.3% for the respective feedstuffs. Effective degradabilities of the OM were 55.7, 56.4 and 47.4%, respectively. Results also showed that effective degradabilities of the ADF were 55, 56.4 and 37.6, respectively. According to the results the researchers concluded that canola and soybean were more degradable in the rumen of the sheep while cottonseed meal were less degradable and, hence resulted in higher rumen undegradable protein.
The main objective of the current work was to evaluate the suitability of ultrasound-assisted extraction (UAE) for the recovery of oil from papaya seed as compared to conventional extraction techniques (i.e., Soxhlet extraction (SXE) and solvent extraction (SE)). In the present study, the recovery yield, fatty acid composition and triacylglycerol profile of papaya seed oil obtained from different extraction methods and conditions were compared. Results indicated that both solvent extraction (SE, 12 h/25 °C) and ultrasound-assisted extraction (UAE) methods recovered relatively high yields (79.1% and 76.1% of total oil content, respectively). Analysis of fatty acid composition revealed that the predominant fatty acids in papaya seed oil were oleic (18:1, 70.5%-74.7%), palmitic (16:0, 14.9%-17.9%), stearic (18:0, 4.50%-5.25%), and linoleic acid (18:2, 3.63%-4.6%). Moreover, the most abundant triacylglycerols of papaya seed oil were triolein (OOO), palmitoyl diolein (POO) and stearoyl oleoyl linolein (SOL). In this study, ultrasound-assisted extraction (UAE) significantly (p < 0.05) influenced the triacylglycerol profile of papaya seed oil, but no significant differences were observed in the fatty acid composition of papaya seed oil extracted by different extraction methods (SXE, SE and UAE) and conditions.
The seeds (6.9±0.2% by weight of fruit) of the red-skin rambutan (Nephelium lappaceum L.) contain a considerable amount of crude fat (38.0±4.36%) and thus, the aim of the study was to determine the physico-chemical properties of this fat for potential applications. The iodine and saponification values, and unsaponifiable matter and free fatty acid contents of the seed fat were 50.27 g I2/100g fat, 182.1 mg KOH/g fat, 0.8% and 2.1%, respectively. The fat is pale yellow with a Lovibond color index of 3.1Y+1.1R. The fatty acid profile indicates an almost equal proportion of saturated (49.1%) and unsaturated (50.9%) fatty acids, where oleic (42.0%) and arachidic (34.3%) acids were the most dominant fatty acids. It also contained small amounts of stearic (8.0%), palmitic (4.6%), gadoleic (5.9%), linoleic (2.2%), behenic (2.1%) palmitoleic (0.7%) myristic (0.1%) and erucic (0.1%) acids. HPLC analysis showed that the fat comprised mainly unknown triacylglycerols (TAG) with high retention times indicating they have higher carbon numbers compared with many vegetable oils. The fat has melting and cooling points of 44.2°C and -42.5°C, respectively, making it a semi-solid at room temperature. The solid content at 0°C was 53.5% and the fat melted completely at 40°C. z-Nose analysis showed that the presence of high levels of volatile compounds in red-skin rambutan seed and seed fat.
Dendrobium sonia-28 is an important ornamental orchid in the Malaysian flower industry. However, the genus faces both low germination rates and the risk of producing heterozygous progenies. Cryopreservation is currently the favoured long-term storage method for orchids with propagation problems. Vitrification, a frequently used cryopreservation technique, involves the application of pretreatments and cryoprotectants to protect and recover explants during and after storage in liquid nitrogen. However, cryopreservation may cause osmotic injuries and toxicity to cryopreserved explants from the use of highly concentrated additives, and cellular injuries from thawing, devitrification and ice formation. Reactive oxygen species (ROS), occurring during dehydration and cryopreservation, may also cause membrane damage. Plants possess efficient antioxidant systems such as the superoxide dismutase (SOD) and catalase (CAT) enzymes to scavenge ROS during low temperature stress. In this study, protocorm-like bodies (PLBs) of Dendrobium sonia-28 were assayed for the total protein content, and both SOD and CAT activities, at each stage of a vitrification exercise to observe for deleterious stages in the protocol. The results indicated that cryopreserved PLBs of Dendrobium sonia-28 underwent excessive post-thawing oxidative stress due to decreased levels of the CAT enzyme at the post-thawing recovery stage, which contributed to the poor survival rates of the cryopreserved PLBs.
Enzymatic reactions involving lipases as catalyst in transesterification can be an excellent alternative to produce environmental-friendly biodiesel. In this study, lipase extracted from Cocoa Pod Husk (CPH) and immobilized through cross linked enzyme aggregate (CLEA) technology catalysed the transesterification of Jatropha curcas oil successfully. Face centered central composite design (FCCCD) under response surface methodology (RSM) was used to get the optimal conditions of 3% (w/w) enzyme loading, 4h reaction time and 1:6 oil/ethanol ratio to achieve the highest conversion of free fatty acid and glycerides into biodiesel (93%). The reusability of CLEA-lipase was tested and after seven cycles, the conversion percentage reduced to 58%. The results revealed that CLEA lipase from CPH is a potential catalyst for biodiesel production.
The details of plant lipid metabolism are relatively well known but the regulation of fatty acid production at the protein level is still not understood. Hence this study explores the importance of phosphorylation as a mechanism to control the activity of fatty acid biosynthetic enzymes using low and high oleic acid mesocarps of oil palm fruit (Elaeis guineensis variety of Tenera). Adaptation of neutral loss-triggered tandem mass spectrometry and selected reaction monitoring to detect the neutral loss of phosphoric acid successfully found several phosphoamino acid-containing peptides. These peptides corresponded to the peptides from acetyl-CoA carboxylase and 3-enoyl-acyl carrier protein reductase as identified by their precursor ion masses. These findings suggest that these enzymes were phosphorylated at 20th week after anthesis. Phosphorylation could have reduce their activities towards the end of fatty acid biosynthesis at ripening stage. Implication of phosphorylation in the regulation of fatty acid biosynthesis at protein level has never been reported.
Effects of biological activated carbon (BAC), biological aerated filter (BAF), alum coagulation and Moringa oleifera coagulation were investigated to remove iron and arsenic contaminants from drinking water. At an initial dose of 5 mg/L, the removal efficiency for arsenic and iron was 63% and 58% respectively using alum, and 47% and 41% respectively using Moringa oleifera. The removal of both contaminants increased with the increase in coagulant dose and decrease in pH. Biological processes were more effective in removing these contaminants than coagulation. Compared to BAF, BAC gave greater removal of both arsenic and iron, removing 85% and 74%, respectively. Longer contact time for both processes could reduce the greater concentration of arsenic and iron contaminants. The addition of coagulation (at 5 mg/L dosage) and a biological process (with 15 or 60 min contact time) could significantly increase removal efficiency, and the maximum removal was observed for the combination of alum and BAC treatment (60 min contact time), with 100% and 98.56% for arsenic and iron respectively. The reduction efficiency of arsenic and iron reduced with the increase in the concentration of dissolved organics in the feedwater due to the adsorption competition between organic molecules and heavy metals.
The first phase of toxicity identification evaluation (TIE) method comprised of physic-chemical fractionation steps of pH adjustment, pH adjustment followed by filtration, aeration, extraction with solid phase C18 column (SPE), oxidant reduction with sodium thiosulphate and EDTA chelation was conducted to characterize the toxicants of a Malaysian landfill leachate. The battery of organisms test chosen were freshwater fish (Rasbora sumatrana), freshwater prawn (Macrobrachium lanchesteri) and tomato seed (Lycoperson esculentum). Toxicity reductions at each step were comparable for all test organisms. The major toxicants present in the leachate were found to be mostly basic in nature and precipitable under acidic conditions as well as containing non-polar organic compounds. A small reduction in toxicity was observed when leachate was treated with sodium thiosulphate in oxidant reduction test indicating the presence of oxidizers. The EDTA chelating step did not significantly reduce toxicity in the test organisms suggesting insignificant level of (toxic) metals.
Pretreatment of the high free fatty acid rubber seed oil (RSO) via esterification reaction has been investigated by using a pilot scale hydrodynamic cavitation (HC) reactor. Four newly designed orifice plate geometries are studied. Cavities are induced by assisted double diaphragm pump in the range of 1-3.5 bar inlet pressure. An optimised plate with 21 holes of 1mm diameter and inlet pressure of 3 bar resulted in RSO acid value reduction from 72.36 to 2.64 mg KOH/g within 30 min of reaction time. Reaction parameters have been optimised by using response surface methodology and found as methanol to oil ratio of 6:1, catalyst concentration of 8 wt%, reaction time of 30 min and reaction temperature of 55°C. The reaction time and esterified efficiency of HC was three fold shorter and four fold higher than mechanical stirring. This makes the HC process more environmental friendly.
Valorizing neglected crop byproducts through eco-friendly techniques has gained attention as underutilized crops offer new sources of bioactive components. This study examined the chemical composition, phytochemicals, and antioxidant activities in six fractions of the underutilized chayote (Sechium edule). Each fraction was microwave-dried and extracted with ultrasound assistance. Shoots were high in ash and fiber, while pulp had high moisture and carbohydrates, and seeds were rich in fat and protein. Shoots, leaves, roots, and peels had high mineral levels, particularly Ca, K, Mg, P, Fe, and Zn. Shoots were also high in phenolics, flavonoids, and ascorbic acid, while leaves had a high carotenoid content. Polyphenol compounds such as myricetin and catechin were abundant in shoots, which exhibited the highest antioxidant activity. Chayote shoots, roots, and leaves are abundant in bioactive compounds that can be utilized in food and, therefore, offers an opportunity for utilization of a currently neglected plant byproduct.
Rambutan seed is usually discarded during fruit processing. However, the seed contains a considerable amount of crude fat. Hence, the objective of this study was to investigate the fat properties and antinutrient content of the seed during fermentation of rambutan fruit. Results showed that the crude fat content of the seed reduced by 22% while its free fatty acid content increased by 4.3 folds after 10 days of fermentation. Arachidic acid was selectively reduced and was replaced by linoleic acid from the seventh day of fermentation onwards. Only 14.5% of triacylglycerol remained in the seed fat at the end of fermentation. The complete melting temperature, crystallization onset temperature and solid fat index at 37 °C of the fermented seed fat were higher than that of non-fermented seed fat. The saponin and tannin contents of the seed were reduced by 67% and 47%, respectively, after fermentation.
The stearoyl-acyl-carrier-protein (ACP) desaturase is a plastid-localized enzyme that catalyzes the conversion of stearoyl-ACP to oleoyl-ACP and plays an important role in the determination of the properties of the majority of cellular glycerolipids. Functional characterization of the fatty acid desaturase genes and their specific promoters is a prerequisite for altering the composition of unsaturated fatty acids of palm oil by genetic engineering. In this paper, the specificity and strength of the oil palm stearoyl-ACP desaturase gene promoter (Des) was evaluated in transgenic tomato plants. Transcriptional fusions between 5' deletions of the Des promoter (Des1-4) and the β-glucuronidase (GUS) reporter gene were generated and their expression analyzed in different tissues of stably transformed tomato plants. Histochemical analysis of the Des promoter deletion series revealed that GUS gene expression was confined to the tomato fruits. No expression was detected in vegetative tissues of the transgenic plants. The highest levels of GUS activity was observed in different tissues of ripe red fruits (vascular tissue, septa, endocarp, mesocarp and columella) and in seeds, which harbored the promoter region located between -590 and +10. A comparison of the promoter-deletion constructs showed that the Des4 promoter deletion (314bp) produced a markedly low level of GUS expression in fruits and seeds. Fluorometric analysis of the GUS activity revealed a 4-fold increase in the activity of the full-length Des promoter compared to the CaMV35S promoter. RNA-hybridization analyses provided additional evidence of increased GUS expression in fruits driven by a Des fragment. Taken together, these results demonstrate the potential of the Des promoter as a tool for the genetic engineering of oil palms and other species, including dicots, in improving the quality and nutritional value of the fruits.
Kenaf (Hibiscus cannabinus L.) is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6-8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.
Dysregulated metabolism is implicated in obesity and other disease conditions like type 2 diabetes mellitus and cardiovascular diseases, which are linked to abnormalities of peroxisome proliferator-activated receptor gamma (PPARγ). PPARγ has been the focus of much research aimed at managing these diseases. Also, germinated brown rice (GBR) is known to possess antidiabetic, antiobesity and hypocholesterolemic effects. We hypothesized that GBR bioactive compounds may mediate some of the improvements in metabolic indices through PPARγ modulation. Cultured HEP-G2 cells were treated with 50 ppm and 100 ppm of extracts from GBR (GABA, ASG and oryzanol) after determination of cell viabilities using MTT assays. Results showed that all extracts upregulated the expression of the PPARγ. However, combination of all three extracts showed downregulation of the gene, suggesting that, in combination, the effects of these bioactives differ from their individual effects likely mediated through competitive inhibition of the gene. Upregulation of the gene may have therapeutic potential in diabetes mellitus and cardiovascular diseases, while its downregulation likely contributes to GBR's antiobesity effects. These potentials are worth studying further.
Radiation processing has been employed successfully for value addition of food and agricultural products. Preliminary studies were undertaken to evaluate the changes induced by ionizing radiation (up to 30 kGy), in the form of gamma irradiation and electron beam irradiation, on some quality attributes and nutritive values of nutraceutically valued lotus seeds. Significant loss in seed firmness was recorded between control and irradiated seeds, irrespective of radiation source. Similarly, the specific viscosity of irradiated lotus seeds decreased significantly up to a dose of 7.5 kGy. Starch increased after exposure to gamma or electron beam irradiation, whereas the total phenolic contents were decreased. Gamma irradiation revealed an enhancement in protein, while the electron beam showed a decrease. Partial oxidation of the seeds during radiation treatments might have occurred as evidenced from the decomposition profiles (thermogravimetry) during heating. It is evident that ionizing radiation brought about significant and variable changes in the quality and nutritive values of lotus seed. Further exploration of this technology for safety and quality is warranted.
Chemoprevention has become an important area in cancer research due to the failure of current therapeutic modalities. Epidemiological and preclinical studies have demonstrated that nutrition plays a vital role in the etiology of cancer. This study was conducted to determine the chemopreventive effects of germinated brown rice (GBR) in rats induced with colon cancer. GBR is brown rice that has been claimed to be richer in nutrients compared to the common white rice. The male Sprague Dawley rats (6 weeks of age) were randomly divided into 5 groups: (G1) positive control (with colon cancer, unfed with GBR), (G2) fed with 2.5 g/kg of GBR (GBR (g)/weight of rat (kg)), (G3) fed with 5 g/kg of GBR, (G4) fed with 10 g/kg of GBR and (G5) negative control (without colon cancer, unfed with GBR). GBR was administered orally once daily via gavage after injection of 15 mg/kg of body weight of azoxymethane (AOM) once a week for two weeks, intraperitonially. After 8 weeks of treatment, animals were sacrificed and colons were removed. Colonic aberrant crypt foci (ACF) were evaluated histopathologically. Total number of ACF and AC, and multicrypt of ACF, and the expression of beta-catenin and COX-2 reduced significantly (p < 0.05) in all the groups treated with GBR (G2, G3 and G4) compared to the control group (G1). Spearman rank correlation test showed significant positive linear relationship between total beta-catenin and COX-2 score (Spearman's rho = 0.616, p = 0.0001). It is demonstrated that GBR inhibits the development of total number of ACF and AC, and multicrypt of ACF, reduces the expression of beta-catenin and COX-2, and thus can be a promising dietary supplement in prevention of colon cancer.
BACKGROUND: We evaluated glycaemic response of a brown rice variant (BR) developed by cross-breeding. Subjects (n = 9) consumed 50 g carbohydrate equivalents of BR, white rice (WR) and the polished brown rice (PR) in comparison to 50 g glucose reference (GLU) in a cross-over design. Plasma glucose and insulin at 0, 15, 45, 60, 90, 120 and 180 min were measured and incremental area under the curve (IAUC) and indices for glucose (GI) and insulin (II) calculated.
RESULTS: BR compared to PR or WR produced the lowest postprandial glycaemia (GI: 51 vs 79 vs 86) and insulinaemia (II: 39 vs 63 vs 68) irrespective of amylose content (19 vs 23 vs 26.5%). Only BR was significantly different from GLU for both plasma glucose (P = 0.012) and insulin (P = 0.013) as well as IAUC(glu) (P = 0.045) and IAUC(ins) (P = 0.031). Glycaemic and insulinaemic responses correlated positively (r = 0.550, P < 0.001). Linear trends for IAUC(glu) and IAUC(ins) indicated a greater secretion of insulin tied in with a greater glycaemic response for WR (r(2) = 0.848), moderate for PR (r(2) = 0.302) and weakest for BR (r(2) = 0.122).
CONCLUSION: The brown rice variant had the lowest GI and II values but these advantages were lost with polishing.
Palm kernel cake (PKC) is a useful source of protein and energy for livestock. Recently, it has been used as an ingredient in poultry feed. Mycotoxin contamination of PKC due to inappropriate handling during production and storage has increased public concern about economic losses and health risks for poultry and humans. This concern has accentuated the need for the evaluation of mycotoxins in PKC. Furthermore, a method for quantifying mycotoxins in PKC has so far not been established. The aims of this study were therefore (1) to develop a method for the simultaneous determination of mycotoxins in PKC and (2) to validate and verify the method. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using an electrospray ionisation interface (ESI) in both positive- and negative-ion modes was developed for the simultaneous determination of aflatoxins (AFB₁, AFB₂, AFG₁ and AFG₂), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB₁ and FB₂), T-2 and HT-2 toxin in PKC. An optimum method using a 0.2 ml min⁻¹ flow rate, 0.2% formic acid in aqueous phase, 10% organic phase at the beginning and 90% organic phase at the end of the gradient was achieved. The extraction of mycotoxins was performed using a solvent mixture of acetonitrile-water-formic acid (79:20:1, v/v) without further clean-up. The mean recoveries of mycotoxins in spiked PKC samples ranged from 81% to 112%. Limits of detection (LODs) and limits of quantification (LOQs) for mycotoxin standards and PKC samples ranged from 0.02 to 17.5 μg kg⁻¹ and from 0.06 to 58.0 μg kg⁻¹, respectively. Finally, the newly developed method was successfully applied to PKC samples. The results illustrated the fact that the method is efficient and accurate for the simultaneous multi-mycotoxin determination in PKC, which can be ideal for routine analysis.