The objective of the study was to investigate the hypoglycaemic properties of Malaysian cocoa (Theobroma cacao) polyphenols extract in-vivo and insulin sensitivity in-vitro. Cocoa extract (CE) (containing 190 - 286 mg total polyphenol per gram extract) was prepared from fermented and roasted (140°C, 20 min) beans by extracting with 80% ethanol in the ratio of 1 to 10. For the in-vivo study, the CE was administered in three dosages (1%, 2%, and 3%) to groups of normal and diabetic rats for a period of 4 weeks by forcefeeding. Results showed that dosages of 1% and 3% CE significantly reduced (p < 0.05) plasma glucose levels in the diabetic rats. An in-vitro study (BRIN-BD11 cell lines) was used to evaluate the effect of CE on insulinsensitivity. The results demonstrated that CE at a concentration of 0.1 mg/ml significantly increased (p < 0.05) insulin level compared to the control. The results of this study showed that Malaysian cocoa polyphenol extract have the potential of being an insulin-mimetic agent. Further studies are on-going to elucidate the underlying mechanisms of polyphenols present in CE that contribute to the reduction of plasma glucose levels and insulin mimicking activity.
Optimization of decolorization of methylene blue (MB) dye by lignin peroxidase (LiP) enzyme produced by white-rot fungus Phanerochaete chrysosporium using sewage treatment plant (STP) sludge as a major substrate was carried out in the laboratory. Optimization by the one-factor-at-a-time (OFAT) and statistical approach was carried out to determine the process conditions on optimum decolorization of MB dye using LiP enzyme in static mode. The OFAT method indicated that the optimum conditions for decolorization of MB dye (removal: 14-40%) was at temperature 55 degrees C, pH 5.0 with hydrogen peroxide (H(2)O(2)) concentration 4.0mM, MB dye concentration 20mg/L and LiP activity 0.487U/ml. The addition of veratryl alcohol to the reaction mixtures did not contribute any further increases in decolorization. The initial concentration of MB and the activity of LiP enzyme were further optimized using response surface methodology (RSM). The contour and surface plots suggested that the optimum initial concentration of MB and LiP activity predicted were 15mg/L and 0.687U/ml, respectively for the removal of 65%. The validation of the model showed that the decolorization process gave the higher removal of 90% in agitation mode compared to the static mode with 65% for 60min of incubation time by LiP enzyme.
The production of xylanase from palm kernel cake as a substrate was studied in solid substrate fermentation. The simultaneous effects of three independent variables, namely incubation temperature, initial moisture content of substrate and air flow rate on xylanase production were evaluated by response surface methodology using central composite face centered design. A total of 18 experiments were carried out in which Aspergillus niger FTCC 5003 was cultivated on palm kernel cake in a column bioreactor for 7 days under incubation temperature, moisture level and aeration rate determined. Test results showed that the highest xylanase activity of 174.88 U g(-1) was produced at incubation temperature, initial moisture level and aeration rate of 25 degrees C, 60% and 1.5 L min(-1), respectively. The statistical analysis of the experimental results revealed that the linear effect of incubation temperature and quadratic term of initial moisture content had highly significant effects on xylanase production (p<0.01). Statistical results also showed that interaction effect between incubation temperature and initial moisture content as well as interaction effect between moisture level and aeration rate influenced the yield ofxylanase at probability levels of 95%. Optimum conditions determined by statistical model for attaining maximum xylanase production were incubation temperature of 25 degrees C, initial moisture level of 63% and aeration rate of 1.76 L min(-1). The xylanase activity of 192.50 U g(-1) was obtained when solid substrate fermentation was performed under the optimal circumstances.
Photo fermentation is a biological process that can be applied for hydrogen production. The process is environmental friendly which is operated under mild conditions using renewable resources. In order to increase yield of H2 produced by Rhodobacter sphaeroides, some experimental factors that may enhance H2 production were studied. The effect of operating parameters including agitation, aeration and light on hydrogen production using R. sphaeroides NCIMB 8253 was investigated. Rhodobacter sphaeroides NCIMB 8253 was grown in 100 mL serum bottle containing growth medium with maliec acid as the sole organic carbon source. The cultures were incubated anaerobically at 30 degrees C with tungsten lamp (100 W) as the light source (3.8 klux) and argon gas was purged for maintaining anaerobic condition. The results show that maximum hydrogen produced was higher (54.37 mL) in static culture with 69.98% of H2 in the total gas compared with shake culture (11.57 mL) with 57.86% of H2. By using static culture, H2 produced was five times higher compared with non-static in both aerobic and anaerobic condition. It was found that growth and H2 production with fluorescent lamp showed better results than growth and H2 production with tungsten light.
To assess the probiotic effects of Lactobacillus agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230 and the pH on the cecal microflora of chicken and metabolic end products.
A two-stage fermentation process consisting of dark and photo-fermentation periods was carried out in a batch reactor. In the first stage, glucose was fermented in the dark stage using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564; CSN1-4) to produce acetate, CO2 and H2. The acetate produced in the first stage is fermented to H2 and CO2 by Rhodobacter sphaeroides NCIMB 8253 for further hydrogen production in the second, illuminated stage. The yield of hydrogen in the first stage was about 3.10 mol H2 (mol glucose)(-1) at a glucose concentration of 10 g L(-1), pH 6 +/- 0.2 and 37 degrees C and the second stage yield was about 1.10-1.25 mol H2 (mol acetic acid)(-1) at pH 6.8 +/- 0.2 and 32 degrees C, without removal of the Clostridium CSN1-4. The overall yield of hydrogen in the two-stage process, with glucose as the main substrate was higher than single-stage fermentation.
The antioxidant properties of virgin coconut oil produced through chilling and fermentation were investigated and compared with refined, bleached and deodorized coconut oil. Virgin coconut oil showed better antioxidant capacity than refined, bleached and deodorized coconut oil. The virgin coconut oil produced through the fermentation method had the strongest scavenging effect on 1,1-diphenyl-2-picrylhydrazyl and the highest antioxidant activity based on the beta-carotene-linoleate bleaching method. However, virgin coconut oil obtained through the chilling method had the highest reducing power. The major phenolic acids detected were ferulic acid and p-coumaric acid. Very high correlations were found between the total phenolic content and scavenging activity (r=0.91), and between the total phenolic content and reducing power (r=0.96). There was also a high correlation between total phenolic acids and beta-carotene bleaching activity. The study indicated that the contribution of antioxidant capacity in virgin coconut oil could be due to phenolic compounds.
Unprocessed ‘budu’ is a mixture of anchovies and salt that has been fermented for a period of time, and has not been heat-treated nor formulated with additional ingredients. This study analyzed Malaysian
unprocessed ‘budu’ from 12 producers for microbiological, salt, protein, histamine and 3-MCPD contents.
The results demonstrated that Malaysian unprocessed ‘budu’ were free from pathogenic Coliform, E. coli,
V. parahaemolyticus and V. cholerae contaminations. Carcinogenic 3-MCPD was below detection level of 2 ppb for all 12 samples tested. However, 58% of the unprocessed ‘budu’ had histamine content greater than the hazardous levels of 50 mg/100 g sample.
The effects of lactic acid bacteria (Lactobacillus plantarum and Lactobacillus bulgaricus) inoculation
on the sensory attributes and consumers acceptance of fermented curry paste compared with uncultured sample were assessed. pH, titratable acidity (TA) and color changes, during four-month storage were monitored. Hedonic test was utilized to evaluate consumer perception and acceptability of fermented and ordinary curry pastes. Rapid pH drop was observed in inoculated sample with Lb. plantarum presenting better performance than the Lb. bulgaricus. Titratable acidity increased significantly (p0.05) in most of the attributes of
original recipe and fermented curry paste except for color and sweetness. In summary, this study showed fermented curry paste with Lb. plantarum and Lb. bulgaricus exhibited new sensory attributes encouraging acceptability by consumers.
In this study, an anaerobic mesophilic bacterial strain, namely Clostridium butyricum KBH1, was isolated from a natural source. This strain grew well and produced biogas with an average hydrogen concentration of 60% (v/v) in the Reinforced Clostridial Media (RCM). To study the basic nutrient requirements, three main nutrients namely peptone (Pep), yeast extracts (Yes) and glucose (Glu) were chosen as factors, using an experimental design. The experiments were run according to 23 Full Factorial Design, followed by the Response Surface Method (RSM). The fermentation was performed in 30 ml serum bottles with 20 ml working volume in a sterile and anaerobic condition at 37°C with 5% inoculums. The results from the Analysis of Variance (ANOVA) for the factorial design showed that all the three factors had significantly affected the gas production by the C. butyricum. The response surface plot of the gas production by C. butyricum showed that the gas production could be enhanced by increasing peptone and yeast extract concentrations up to 15 g/l and 24 g/l respectively, without showing any substrate inhibition. Meanwhile, the glucose concentration showed an optimum at the middle point (8 g/l) with possible substrate inhibition at a high concentration (12 g/l). The total biogas production could be correlated to the three factors, using the quadratic equation: Gas =0.17 + 7.11Glu - 0.02Pep + 0.77Yes - 0.53Glu2 + 0.09Glu*Pep. The experimental results showed that the strain could grow well in substrate with high organic nitrogen content such as POME and might be not suitable for substrate with high sugar content due to substrate inhibition.
Streptococcus zooepidemicus (SZ) is an aerotolerant bacteria and its ability to survive under reactive oxidant raises the question of the existence of a defense system against oxidative stress. As a characteristic of lactic acid bacteria, Streptococcus lacks an ordinary anti-oxidative stress enzyme, catalases and an electron transport chain. Whether this bacterium resists oxidative stress prior to an exposure to a higher level of an oxidizing agent H2O2 in hyaluronic acid fermentation is not known. This paper describes that Streptococcus cells, once treated with lower concentrations of H2O2 (i.e. 0.25, 0.50 and 1.0 mM) at least, were prepared for a subsequent higher concentrations of H2O2 such as 20.5 and 100 mM. At low concentrations (i.e. 0.25, 0.50 and 1.0 mM), H2O2 was found to act as a stimulant for HA synthesis, but it became toxic if presented at a very high level (100 mM H2O2). The highest HA yield to glucose consumed (YHAtotal/glu) was 0.017 gg-1 for the cells pre-treated with 0 mM of H2O2, and then exposed to 20.5 mM H2O2. Thus, this implied that this bacteria might possess a defense mechanism against oxidative stress and that this system was inducible.
The production of ethanol, from glucose in batch and fed batch culture, was investigated. In the fed batch culture, the glucose feeding was added into the culture at 16th hour of fermentation. The effects of different glucose concentration feeding rates on ethanol fermentation were investigated for fed batch culture. The 2gL-1hr-1 glucose concentration feeding rate was found to give higher ethanol yield (2.47 g ethanol g glucose-1), with respect to substrate consumed as compared to 8 gL-1hr-1 (0.23 g ethanol g glucose-1) and 4 gL-1hr-1 (0.20 g ethanol g glucose-1). The ethanol yield with respect to substrate consumed obtained in batch culture was 0.81 g ethanol g glucose-1. The fed batch culture at 2 gL-1hr-1 glucose concentration feeding rate was proven to be a better fermentation system than the batch culture. The specific growth rate, specific glucose consumption rate and specific ethanol production rate for the fed batch fermentation, at 2 gL-1hr-1 glucose concentration feeding rate, were 0.065 hr-1, 1.20 hr-1 and 0.0009 hr-1, respectively.
Acetone-butanol-ethanol (ABE) fermentation from Palm Oil Mill Effluent (POME) by C. acetobutylicum NCIMB 13357 in an oscillatory flow bioreactor was investigated. Experimental works were conducted in a U-shaped stainless steel oscillatory flow bioreactor at oscillation frequency between 0.45-0.78 Hz and a constant amplitude of 12.5 mm. Fermentations were carried out for 72 hr at 35oC using palm oil mill effluent and reinforced clostridia medium as a growth medium in batch culture. Result of this investigation showed that POME is a viable media for ABE fermentation and oscillatory flow bioreactor has an excellent potential as an alternative fermentation device.
The production of beta-mannanase from palm kernel cake (PKC) as a substrate in solid substrate fermentation (SSF) was studied using a laboratory column bioreactor. The simultaneous effects of three independent variables, namely incubation temperature, initial moisture content of substrate and airflow rate, on beta-mannanase production were evaluated by response surface methodology (RSM) on the basis of a central composite face-centered (CCF) design. Eighteen trials were conducted in which Aspergillus niger FTCC 5003 was cultivated on PKC in an aerated column bioreactor for seven days under SSF process. The highest level of beta-mannanase (2117.89 U/g) was obtained when SSF process was performed at incubation temperature, initial moisture level and aeration rate of 32.5 degrees C, 60% and 0.5 l/min, respectively. Statistical analysis revealed that the quadratic terms of incubation temperature and initial moisture content had significant effects on the production of beta-mannanase (P < 0.01). A similar analysis also demonstrated that the linear effect of initial moisture level and an interaction effect between the initial moisture content and aeration rate significantly influenced the production of beta-mannanase (P < 0.01). The statistical model suggested that the optimal conditions for attaining the highest level of beta-mannanase were incubation temperature of 32 degrees C, initial moisture level of 59% and aeration rate of 0.5 l/min. A beta-mannanase yield of 2231.26 U/g was obtained when SSF process was carried out under the optimal conditions described above.
The in vitro fermentability of sago (Metroxylon sagu) resistant starch type III (RS(3)) by selected probiotic bacteria was investigated. Sago RS(3) with 12% RS content was prepared by enzymatic debranching of native sago starch with pullulanase enzyme, followed by autoclaving, cooling, and annealing. The fermentation of sago RS(3) by L. acidophilus FTCC 0291, L. bulgaricus FTCC 0411, L. casei FTCC 0442, and B. bifidum BB12 was investigated by observing the bacterial growth, carbohydrate consumption profiles, pH changes, and total short chain fatty acids (SCFA) produced in the fermentation media. Comparisons were made with commercial fructo-oligosaccharide (FOS), Hi-maize 1043, and Hi-maize 240. Submerged fermentations were conducted in 30 mL glass vials for 24 h at 37 degrees C in an oven without shaking. The results indicated that fermentation of sago RS(3) significantly (P < 0.05) yielded the highest count of Lactobacillus sp. accompanied by the largest reduction in pH of the medium. Sago RS(3) was significantly the most consumed substrate compared to FOS and Hi-maizes.
Lactobacillus sp. FTDC 2113, L. acidophilus FTDC 8033, L. acidophilus ATCC 4356, L. casei ATCC 393, Bifidobacterium FTDC 8943 and B. longum FTDC 8643 were incorporated into soymilk supplemented with fructooligosaccharides (FOS), inulin, mannitol, maltodextrin and pectin. The objective of the present study was to evaluate the effects of prebiotics on the bioactivity of probiotic-fermented soymilk. Proteolytic activity was increased in the presence of FOS, while the supplementation of inulin and pectin increased the angiotensin I-converting enzyme inhibitory activity accompanied by lower IC(50) values. The beta-glucosidase activity was also enhanced in the presence of pectin. This led to higher bioconversion of glucosides to aglycones by probiotics, especially genistin and malonyl genistin to genistein. Results from this study indicated that the supplementation of prebiotics enhanced the in-vitro antihypertensive effect and production of bioactive aglycones in probiotic-fermented soymilk. Therefore, this soymilk could potentially be used as a dietary therapy to reduce the risks of hypertension and hormone-dependent diseases such as breast cancer, prostate cancer and osteoporosis.
Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. The objective of this study was to evaluate the effects of agrowastes from durian (Durio zibethinus), cempedak (Artocarpus champeden), and mangosteen (Garcinia mangostana) as immobilizers for lactobacilli grown in soymilk. Rinds from the agrowastes were separated from the skin, dried, and ground (150 microm) to form powders and used as immobilizers. Scanning electron microscopy revealed that lactobacilli cells were attached and bound to the surface of the immobilizers. Immobilized cells of Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and L. bulgaricus FTCC 0411 were inoculated into soymilk, stored at room temperature (25 degrees C) and growth properties were evaluated over 168 h. Soymilk inoculated with nonimmobilized cells was used as the control. Utilization of substrates, concentrations of lactic and acetic acids, and changes in pH were evaluated in soymilk over 186 h. Immobilized lactobacilli showed significantly better growth (P < 0.05) compared to the control, accompanied by higher production of lactic and acetic acids in soymilk. Soymilk containing immobilized cells showed greater reduction of soy sugars such as stachyose, raffinose, sucrose, fructose, and glucose compared to the control (P < 0.05).
Ethanolic fermentation using Saccharomyces cerevisiae was carried out on three types of hydrolysates produced from lignocelulosic biomass which are commonly found in Malaysia such as oil palm trunk, rubberwood and mixed hardwood. The effect of fermentation temperature and pH of hydrolysate was evaluated to optimize the fermentation efficiency which defined as maximum ethanol yield in minimum fermentation time. The fermentation process using different temperature of 25 degrees Celsius, 30 degrees Celsius and 40 degrees Celsius were performed on the prepared fermentation medium adjusted to pH 4, pH 6 and pH 7, respectively. Results showed that the fermentation time was significantly reduced with the increase of temperature but an adverse reduction in ethanol yield was observed using temperature of 40 degrees Celsius. As the pH of hydrolysate became more acidic, the ethanol yield increased. Optimum fermentation efficiency for ethanolic fermentation of lignocellulosic hydrolysates using S. cerevisiae can be obtained using 33.2 degrees Celsius and pH 5.3.
Box-Wilson design (BWD) model was applied to determine the optimum values of influencing parameters in anaerobic fermentation to produce hydrogen using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). The main focus of the study was to find the optimal relationship between the hydrogen yield and three variables including initial substrate concentration, initial medium pH and reaction temperature. Microbial growth kinetic parameters for hydrogen production under anaerobic conditions were determined using the Monod model with incorporation of a substrate inhibition term. The values of micro(max) (maximum specific growth rate) and K, (saturation constant) were 0.398 h(-1) and 5.509 g L(-1), respectively, using glucose as the substrate. The experimental substrate and biomass-concentration profiles were in good agreement with those obtained by the kinetic-model predictions. By varying the conditions of the initial substrate concentration (1-40 g L(-1)), reaction temperature (25-40 degrees C) and initial medium pH (4-8), the model predicted a maximum hydrogen yield of 3.24 mol H2 (mol glucose)(-1). The experimental data collected utilising this design was successfully fitted to a second-order polynomial model. An optimum operating condition of 10 g L(-1) initial substrate concentration, 37 degrees C reaction temperature and 6.0 +/- 0.2 initial medium pH gave 80% of the predicted maximum yield of hydrogen where as the experimental yield obtained in this study was 77.75% exhibiting a close accuracy between estimated and experimental values. This is the first report to predict bio-hydrogen yield by applying Box-Wilson Design in anaerobic fermentation while optimizing the effects of environmental factors prevailing there by investigating the effects of environmental factors.