The energy-salvaging capacity of the gut microbiota from dietary ingredients has been proposed as a contributing factor for the development of obesity. This knowledge generated interest in the use of non-digestible dietary ingredients such as prebiotics to manipulate host energy homeostasis. In the present study, the in vitro response of obese human faecal microbiota to novel oligosaccharides was investigated. Dextrans of various molecular weights and degrees of branching were fermented with the faecal microbiota of healthy obese adults in pH-controlled batch cultures. Changes in bacterial populations were monitored using fluorescent in situ hybridisation and SCFA concentrations were analysed by HPLC. The rate of gas production and total volume of gas produced were also determined. In general, the novel dextrans and inulin increased the counts of bifidobacteria. Some of the dextrans were able to alter the composition of the obese human microbiota by increasing the counts of Bacteroides-Prevotella and decreasing those of Faecalibacterium prausnitzii and Ruminococcus bromii/R. flavefaciens. Considerable increases in SCFA concentrations were observed in response to all substrates. Gas production rates were similar during the fermentation of all dextrans, but significantly lower than those during the fermentation of inulin. Lower total gas production and shorter time to attain maximal gas production were observed during the fermentation of the linear 1 kDa dextran than during the fermentation of the other dextrans. The efficacy of bifidobacteria to ferment dextrans relied on the molecular weight and not on the degree of branching. In conclusion, there are no differences in the profiles between the obese and lean human faecal fermentations of dextrans.
Lignocellulosic biomass is a potential substrate for ethanol production. However, pretreatment of lignocellulosic materials produces inhibitory compounds such as acetic acid, which negatively affect ethanol production by Saccharomyces cerevisiae. Supplementation of the medium with three metal ions (Zn(2+) , Mg(2+) , and Ca(2+) ) increased the tolerance of S. cerevisiae toward acetic acid compared to the absence of the ions. Ethanol production from xylose was most improved (by 34%) when the medium was supplemented with 2 mM Ca(2+) , followed by supplementation with 3.5 mM Mg(2+) (29% improvement), and 180 μM Zn(2+) (26% improvement). Higher ethanol production was linked to high cell viability in the presence of metal ions. Comparative transcriptomics between the supplemented cultures and the control suggested that improved cell viability resulted from the induction of genes controlling the cell wall and membrane. Only one gene, FIT2, was found to be up-regulated in common between the three metal ions. Also up-regulation of HXT1 and TKL1 might enhance xylose consumption in the presence of acetic acid. Thus, the addition of ionic nutrients is a simple and cost-effective method to improve the acetic acid tolerance of S. cerevisiae.
Rice straw has shown to be a promising agricultural by-product in the bioconversion of biomass to value-added products. Hydrolysis of cellulose, a main constituent of lignocellulosic biomass, is a requirement for fermentable sugar production and its subsequent bioconversion to biofuels such as biobutanol. The high cost of commercial enzymes is a major impediment to the industrial application of cellulases. Therefore, the use of local microbial enzymes has been suggested. Trichoderma harzianum strains are potential CMCase and β-glucosidase producers. However, few researches have been reported on cellulase production by T. harzianum and the subsequent use of the crude cellulase for cellulose enzymatic hydrolysis. For cellulose hydrolysis to be efficiently performed, the presence of the whole set of cellulase components including exoglucanase, endoglucanase, and β-glucosidase at a considerable concentration is required. Biomass recalcitrance is also a bottleneck in the bioconversion of agricultural residues to value-added products. An effective pretreatment could be of central significance in the bioconversion of biomass to biofuels.
Biosurfactants are surface-active compounds produced by different microorganisms. The aim of this study was to introduce palm kernel cake (PKC) as a novel substrate for biosurfactant production using a potent bacterial strain under liquid state fermentation. This study was primarily based on the isolation and identification of biosurfactant-producing bacteria that could utilize palm kernel cake as a new major substrate. Potential bacterial strains were isolated from degraded PKC and screened for biosurfactant production with the help of the drop collapse assay and by analyzing the surface tension activity. From the screened isolates, a new strain, SM03, showed the best and most consistent results, and was therefore selected as the most potent biosurfactant-producing bacterial strain. The new strain was identified as Providencia alcalifaciens SM03 using the Gen III MicroPlate Biolog Microbial Identification System. The yield of the produced biosurfactant was 8.3 g/L.
An innovative approach using soybean residues for the production of bioflocculants through solid-state fermentation was carried out in 4.5 L near-to-adiabatic bioreactors at pilot-scale level. An added inoculum of the strain Bacillus subtilis UPMB13 was tested in comparison with control reactors without any inoculation after the thermophilic phase of the fermentation. The flocculating performances of the extracted bioflocculants were tested on kaolin suspensions, and crude bioflocculants were obtained from 20 g of fermented substrate through ethanol precipitation. The production of bioflocculants was observed to be higher during the death phase of microbial growth. The bioflocculants were observed to be granular in nature and consisted of hydroxyl, carboxyl and methoxyl groups that aid in their flocculating performance. The results show the vast potential of the idea of using wastes to produce bioactive materials that can replace the current dependence on chemicals, for future prospect in water treatment applications.
The biosynthesis of biomedical products including lipid and gamma-linolenic acid (GLA) by Cunninghamella bainieri 2A1 was studied in repeated batch fermentation. Three key process variables, namely, glucose concentration, ammonium tartrate concentration, and harvesting time, were optimized using response surface methodology. Repeated batch fermentation was carried out by the cultivation of Cunninghamella bainieri 2A1 in nitrogen-limited medium with various nitrogen concentration (1-4 g/L) and glucose concentration (20-40 g/L) at three time intervals (12 h, 24 h, and 48 h). Experimental results showed that the highest lipid concentration of 6.2 g/L and the highest GLA concentration of 0.4 g/L were obtained in optimum conditions, where 20.2 g/L glucose, 2.12 g/L ammonium tartrate, and 48 h harvesting time were utilized. Statistical results showed that the interaction between glucose and ammonium tartrate concentration had highly significant effects on lipid and GLA biosynthesis (P < 0.01). Moreover, harvesting time had a significant interaction effect with glucose and ammonium tartrate concentration on lipid production (P < 0.05).
Effective optimization of microalgae-to-bioethanol process systems hinges on an in-depth characterization of key process parameters relevant to the overall bioprocess engineering. One of the such important variables is the biomass particle size distribution and the effects on saccharification levels and bioethanol titres. This study examined the effects of three different microalgal biomass particle size ranges, 35 μm ≤ x ≤ 90 μm, 125 μm ≤ x ≤ 180 μm, and 295 μm ≤ x ≤ 425 μm, on the degree of enzymatic hydrolysis and bioethanol production. Two scenarios were investigated: single enzyme hydrolysis (cellulase) and double enzyme hydrolysis (cellulase and cellobiase). The glucose yield from biomass in the smallest particle size range (35 μm ≤ x ≤ 90 μm) was the highest, 134.73 mg glucose/g algae, while the yield from biomass in the larger particle size range (295 μm ≤ x ≤ 425 μm) was 75.45 mg glucose/g algae. A similar trend was observed for bioethanol yield, with the highest yield of 0.47 g EtOH/g glucose obtained from biomass in the smallest particle size range. The results have shown that the microalgal biomass particle size has a significant effect on enzymatic hydrolysis and bioethanol yield.
Species of the genus Ganoderma are a cosmopolitan wood decaying white rot fungi, which has been used by the Asians for therapeutic purposes for centuries. In the present study, solid-substrate fermentation (SSF) of wheat grains (Triticum aestivum L.) was carried out with indigenous Ganoderma australe (KUM60813) and G. neo-japonicum (KUM61076) selected based on ethnomycological knowledge. G. lucidum (VITA GL) (a commercial strain) was also included in the study. Antioxidant activities of the crude ethanol and aqueous extracts of the fermented and unfermented wheat grains were investigated by ferric reducing antioxidant power (FRAP), Trolox equivalent antioxidant capacity (TEAC), diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging ability, and lipid peroxidation assay. Among the six mycelia extracts tested, the ethanol extract from wheat fermented with KUM61076 mycelia showed the most potent antioxidant activities, whereas the ethanol extract of wheat grains fermented with KUM60813 mycelia has a good potential in protecting frying oils against oxidation. Total phenolic content (TPC) in the ethanol extracts were higher than that in the aqueous extract. The wheat grains fermented with G. australe (KUM60813) and G. neo-japonicum KUM61076 have greater antioxidant potential compared to the commercially available G. lucidum (VITA GL). The antioxidant activities of the mycelia extracts had a positive correlation with their phenolic contents. Thus phenolic compounds may play a vital role in the antioxidant activities of the selected Ganoderma spp.
Previous studies on the nutritional and nutraceutical properties of Lignosus rhinocerotis focused mainly on the sclerotium; however, the supply of wild sclerotium is limited. In this investigation, the antioxidant capacity and cytotoxic effect of L. rhinocerotis cultured under different conditions of liquid fermentation (shaken and static) were compared to the sclerotium produced by solid-substrate fermentation. Aqueous methanol extracts of the mycelium (LR-MH, LR-MT) and culture broth (LR-BH, LR-BT) demonstrated either higher or comparable antioxidant capacities to the sclerotium extract (LR-SC) based on their radical scavenging abilities, reducing properties, metal chelating activities, and inhibitory effects on lipid peroxidation. All extracts exerted low cytotoxicity (IC50>200 µg/ml, 72 h) against selected mammalian cell lines. Several low-molecular-weight compounds, including sugars, fatty acids, methyl esters, sterols, amides, amino acids, phenolics, and triterpenoids, were identified using GC-MS and UHPLC-ESI-MS/MS. The presence of proteins (<40 kDa) in the extracts was confirmed by SDS-PAGE and SELDI-TOF-MS. Principal component analysis revealed that the chemical profiles of the mycelial extracts under shaken and static conditions were distinct from those of the sclerotium. Results from bioactivity evaluation and chemical profiling showed that L. rhinocerotis from liquid fermentation merits consideration as an alternative source of functional ingredients and potential substitute for the sclerotium.
A pilot-scale production of lipase using palm oil mill effluent (POME) as a fermentation basal medium was carried out, and parameters for immobilization of the produced lipase were optimized. Lipase production in a 300-L bioreactor was performed using two proposed strategies, constant power per volume (P/V) and constant tip speed. Moreover, lipase immobilization on different materials was also investigated. Lipase production was performed using liquid-state bioconversion of POME as the medium and Candida cylindracea as the inoculum. The fermentation medium was composed of 1% total suspended solids (TSS) of POME, 0.5% (w/v) peptone, 0.7% (v/v) Tween-80, and 2.2% inoculum. The medium composition was decided on the basis of the medium optimization results of a previous study. The fermentation was carried out for 48 h at 30°C and pH 6. The maximum lipase production was 5.72U/mL and 21.34 U/mL, obtained from the scale-up strategies of constant tip speed and P/V, respectively. Four accessible support materials were screened for their potential use in immobilization. The most suitable support material was found to be activated carbon, with a maximum immobilization of 94%.
Four cellulolytic and hemicellulolytic bacterial cultures were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). Two experiments were conducted; the objective of the first experiment was to determine the optimum time period required for solid state fermentation (SSF) of palm kernel cake (PKC), whereas the objective of the second experiment was to investigate the effect of combinations of these cellulolytic and hemicellulolytic bacteria on the nutritive quality of the PKC. In the first experiment, the SSF was lasted for 12 days with inoculum size of 10% (v/w) on different PKC to moisture ratios. In the second experiment, fifteen combinations were created among the four microbes with one untreated PKC as a control. The SSF lasted for 9 days, and the samples were autoclaved, dried, and analyzed for proximate analysis. Results showed that bacterial cultures produced high enzymes activities at the 4th day of SSF, whereas their abilities to produce enzymes tended to be decreased to reach zero at the 8th day of SSF. Findings in the second experiment showed that hemicellulose and cellulose was significantly (P < 0.05) decreased, whereas the amount of reducing sugars were significantly (P < 0.05) increased in the fermented PKC (FPKC) compared with untreated PKC.
Oil palm frond (OPF) juice is a potential industrial fermentation substrate as it has high sugars content and the OPF are readily available daily. However, maximum sugars yield and storage stability of the OPF juice are yet to be determined. This study was conducted to determine the effect of physical pretreatment and storage duration of OPF petiole on sugars yield. Storage stability of OPF juice at different storing conditions was also investigated. It was found that OPF petiole squeezed by hydraulic pressing machine gave the highest sugars recovery at almost 40 g/kg, accounting for a recovery yield of 88%. Storage of OPF petiole up to 72 hrs prior to squeezing reduced the free sugars by 11 g/kg. Concentrated OPF juice with 95% water removal had the best storage stability at both 4 and 30°C, when it was stored for 10 days. Moreover, concentrated OPF syrup prepared by thermal processing did not give any Maillard effect on microbial growth. Based on our results, OPF juice meets all the criteria as a good fermentation substrate as it is renewable, consistently available, and easy to be obtained, it does not inhibit microbial growth and product formation, and it contains no impurities.
The intrinsic growth, substrate uptake, and product formation biokinetic parameters were obtained for the anaerobic bacterium, Clostridium ljungdahlii, grown on synthesis gas in various pressurized batch bioreactors. A dual-substrate growth kinetic model using Luong for CO and Monod for H2 was used to describe the growth kinetics of the bacterium on these substrates. The maximum specific growth rate (μ(max) = 0.195 h(-1)) and Monod constants for CO (K s,CO = 0.855 atm) and H2 (K(s,H2) = 0.412 atm) were obtained. This model also accommodated the CO inhibitory effects on cell growth at high CO partial pressures, where no growth was apparent at high dissolved CO tensions (P(CO)(∗) > 0.743 atm). The Volterra model, Andrews, and modified Gompertz were, respectively, adopted to describe the cell growth, substrate uptake rate, and product formation. The maximum specific CO uptake rate (q(max) = 34.364 mmol/g cell/h), CO inhibition constant (K(I) = 0.601 atm), and maximum rate of ethanol (R(max) = 0.172 mmol/L/h at P(CO) = 0.598 atm) and acetate (R(max) = 0.096 mmol/L/h at P(CO) = 0.539 atm) production were determined from the applied models.
Bioethanol production from renewable sources to be used in transportation is now an increasing demand worldwide due to continuous depletion of fossil fuels, economic and political crises, and growing concern on environmental safety. Mainly, three types of raw materials, that is, sugar juice, starchy crops, and lignocellulosic materials, are being used for this purpose. This paper will investigate ethanol production from free sugar containing juices obtained from some energy crops such as sugarcane, sugar beet, and sweet sorghum that are the most attractive choice because of their cost-effectiveness and feasibility to use. Three types of fermentation process (batch, fed-batch, and continuous) are employed in ethanol production from these sugar juices. The most common microorganism used in fermentation from its history is the yeast, especially, Saccharomyces cerevisiae, though the bacterial species Zymomonas mobilis is also potentially used nowadays for this purpose. A number of factors related to the fermentation greatly influences the process and their optimization is the key point for efficient ethanol production from these feedstocks.
The culture conditions for gibberellic acid (GA3) production by the fungus Penicillium variable (P. variable) was optimized using a statistical tool, response surface methodology (RSM). Interactions of culture conditions and optimization of the system were studied using Box-Behnken design (BBD) with three levels of three variables in a batch flask reactor. Experimentation showed that the model developed based on RSM and BBD had predicted GA3 production with R(2) = 0.987. The predicted GA3 production was optimum (31.57 mg GA3/kg substrate) when the culture conditions were at 7 days of incubation period, 21% v/w of inoculum size, and 2% v/w of olive oil concentration as a natural precursor. The results indicated that RSM and BBD methods were effective for optimizing the culture conditions of GA3 production by P. variable mycelia.
The presence of phorbol esters (PEs) with toxic properties limits the use of Jatropha curcas kernel in the animal feed industry. Therefore, suitable methods to detoxify PEs have to be developed to render the material safe as a feed ingredient. In the present study, the biological treatment of the extracted PEs-rich fraction with non-pathogenic fungi (Trichoderma harzianum JQ350879.1, T. harzianum JQ517493.1, Paecilomyces sinensis JQ350881.1, Cladosporium cladosporioides JQ517491.1, Fusarium chlamydosporum JQ350882.1, F. chlamydosporum JQ517492.1 and F. chlamydosporum JQ350880.1) was conducted by fermentation in broth cultures. The PEs were detected by liquid chromatography-diode array detector-electrospray ionization mass spectrometry (LC-DAD-ESIMS) and quantitatively monitored by HPLC using phorbol-12-myristate 13-acetate as the standard. At day 30 of incubation, two T. harzianum spp., P. sinensis and C. cladosporioides significantly (p < 0.05) removed PEs with percentage losses of 96.9%-99.7%, while F. chlamydosporum strains showed percentage losses of 88.9%-92.2%. All fungal strains could utilize the PEs-rich fraction for growth. In the cytotoxicity assay, cell viabilities of Chang liver and NIH 3T3 fibroblast cell lines were less than 1% with the untreated PEs-rich fraction, but 84.3%-96.5% with the fungal treated PEs-rich fraction. There was no inhibition on cell viability for normal fungal growth supernatants. To conclude, Trichoderma spp., Paecilomyces sp. and Cladosporium sp. are potential microbes for the detoxification of PEs.
This study evaluated and compared the antioxidant capacity between freshly prepared and lactic fermented Malaysian herbal teas. Herbal teas are rich in antioxidants. Fermentation has been known to be the oldest and cost effective method with the ability to preserve or improve food nutritional qualities. Information on the antioxidant capacity of lactic fermented food or beverage is still lacking. Hence, the objective of this study is to determine the changes in the antioxidant properties of Malaysian herbal teas after being subjected to lactic fermentation. Commercially available local herbal teas were used for this study. Herbal teas such as “Allspice”, “Scaphium”, “Gora” and “Cinnamon” were purchased from the local store in Malaysia and were subjected to 24-hour lactic fermentation. Lactic fermented herbal teas were analyzed for their total phenolic, total flavonoid and antioxidant properties via DPPH, FRAP, and β-carotene linoleate bleaching assay. All lactic fermented herbal teas exhibited higher phenolic contents, flavonoid contents and antioxidant properties compared to the freshly-prepared herbal teas with majority showing significant changes (p < 0.05) in FRAP and β-carotene bleaching assay. Lactic fermented herbal teas also showed an increase in antioxidant capacity in DPPH assay, however non-significant changes were observed.
The advantage of cooking cannot be summarized just as the better food digestion. Some investigations showed the effect of cooking on reduction of food anti-nutrients such as oxalate. This study was aimed to determine the effect of cooking on oxalate content and its negative effects on calcium availability in eight Malaysian soy-based dishes. Since there is few data which examined the effects of cooking on food oxalate content globally, thus this study was designed as the first in Malaysia. Oxalate in this research was analyzed by using enzymatic methods, while calcium content was determined by using Atomic Absorption Spectrophotometer. The oxalate concentration was in the range of 6.43-19.40 mg/100 g for whole cooked samples, 9.03-11.90 mg/100 g for raw soy products, and 4.36-7.99 mg/100 g for cooked ones. There were 5 out of 12 samples containing oxalate, which was significantly lower (p < 0.05) in cooked products compared to the raw ones. The rest of the samples were also lower in oxalate but not significantly different (p > 0.05). Oxalate in raw/cooked fermented soy products (tempeh) was slightly lower compared to the non-fermented ones. However, there was no significant difference (p > 0.05) in oxalate amount between fermented and non-fermented soy products. As Oxalate/Calcium ratio was below 1, oxalate did not have an effect on availability of calcium in the studied samples. Optimal cooking and food processing might be effective in reducing oxalate content in soy products. There is a need for more investigations about the effect of cooking on soy products to confirm the present results.
In this study, pineapple cannery waste materials were used as substrate for the microbial production of vanillic acid and vanillin by Aspergillus niger I-1472 and Pycnoporus cinnabarinus MUCL 39533. Biotransformation of ferulic acid from pineapple waste by A. niger I-1472 to vanillic acid was optimized using Response Surface Methodology (RSM). A central composite rotatable design was used to allocate treatment combinations and factors tested for their influence on vanillic acid production were inoculum size, yeast extract concentration, diammonium tartrate concentration and initial medium pH. The amount of vanillic acid produced was used as the response for the fermentation study and was assumed to be under the influence of the four factors tested. The estimated conditions for optimal vanillic acid production were inoculum size, 3.08 ×105 CFU mL-1; yeast extract, 0.37 gL-1; diammonium tartrate, 3.88 gL-1 and initial pH, 4.3. Subsequent biotransformation of vanillic acid by P. cinnabarinus MUCL 39533 to vanillin was enhanced with the addition of resin. Under these optimal conditions, 141.00 mgL-1 of vanillin was produced from 5 g of pineapple cannery waste.
Fermented red yeast rice has been traditionally consumed as medication in Asian cuisine. This study aimed to determine the in vivo hypocholesterolemic and antioxidant effects of fermented red yeast rice water extract produced using Malaysian Agricultural Research and Development Institute (MARDI) Monascus purpureus strains in mice fed with high cholesterol diet. Absence of monacolin-k, lower level of γ-aminobutyric acid (GABA), higher content of total amino acids, and antioxidant activities were detected in MARDI fermented red yeast rice water extract (MFRYR). In vivo MFRYR treatment on hypercholesterolemic mice recorded similar lipid lowering effect as commercial red yeast rice extract (CRYR) as it helps to reduce the elevated serum liver enzyme and increased the antioxidant levels in liver. This effect was also associated with the upregulation of apolipoproteins-E and inhibition of Von Willebrand factor expression. In summary, MFRYR enriched in antioxidant and amino acid without monacolin-k showed similar hypocholesterolemic effect as CRYR that was rich in monacolin-k and GABA.