The use of component from Ganoderma lucidum as prebiotic source is interesting as the G. lucidum itself was known for more than a decade in the traditional Chinese medicine. In this work, Ganoderma lucidum crude polysaccharides (GLCP) and Polysaccharide-fraction number 2 (PF-2) were used as carbon sources in the fermentation with Bifidobacterium sp. The results showed the potential of prebiotic effect of the G. lucidum extract in batch-culture fermentation based on increment in the growth of bacteria used (0.4 – 1.5 log10 CFU/mL) after 18h fermentation. Fermentation was further done using faecal materials as bacterial inocula and bacterial growth changes were examined using real-time PCR. The results showed the ability of GLCP and PF-2 to support the growth of Bifidobacterium genus with 0.3 and 0.7 log10 cells/ml increased, respectively. Interestingly, Lactobacillus which is known as beneficial bacterial genus also showed growth increment with 0.7 and 1 log10 cells/ml increased. The competition for carbon sources thus inhibits the growth of potentially harmful genus, Salmonella (0.3 and 0.5 log10 cells/ml) in comparison to the control.
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.
Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [(P(3HB-co-4HB)] copolymer receives attention as next generation biomaterial in medical application. However, the exploitation of the copolymer is still constrained since such copolymer has not yet successfully been performed in industrial scale production. In this work, we intended to establish pilot production system of the copolymer retaining the copolymer quality which has recently discovered to have novel characteristic from lab scale fermentation. An increase of agitation speed has significantly improved the copolymer accumulation efficiency by minimizing the utilization of substrates towards cell growth components. This is evidenced by a drastic increase of PHA content from 28 wt% to 63 wt% and PHA concentration from 3.1 g/L to 6.5 g/L but accompanied by the reduction of residual biomass from 8.0 g/L to 3.8 g/L. Besides, fermentations at lower agitation and aeration have resulted in reduced molecular weight and mechanical strength of the copolymer, suggesting the role of sufficient oxygen supply efficiency in improving the properties of the resulting copolymers. The KLa-based scale-up fermentation was performed successfully in maintaining the yield and the quality of the copolymers produced without a drastic fluctuation. This suggests that the scale-up based on the KLa values supported the fermentation system of P(3HB-co-4HB) copolymer production in single-stage using mixed-substrate cultivation strategy.
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 pretreatment of empty fruit bunch (EFB) was conducted using an integrated system of IL and cellulases (IL-E), with simultaneous fermentation in one vessel. The cellulase mixture (PKC-Cel) was derived from Trichoderma reesei by solid-state fermentation. Choline acetate [Cho]OAc was utilized for the pretreatment due to its biocompatibility and biodegradability. The treated EFB and its hydrolysate were characterized by the Fourier transform infrared spectroscopy, scanning electron microscopy, and chemical analysis. The results showed that there were significant structural changes in EFB after the treatment in IL-E system. The sugar yield after enzymatic hydrolysis by the PKC-Cel was increased from 0.058 g/g of EFB in the crude sample (untreated) to 0.283 and 0.62 ± 06 g/g in IL-E system after 24 and 48 h of treatment, respectively. The EFB hydrolysate showed the eligibility for ethanol production without any supplements where ethanol yield was 0.275 g ethanol/g EFB in the presence of the IL, while lower yield obtained without IL-pretreatment. Moreover, it was demonstrated that furfural and phenolic compounds were not at the level of suppressing the fermentation process.
As fermentation could reduce the negative effects of bran on final cereal products, the utilization of whole-cereal flour is recommended, such as brown rice flour as a functional food ingredient. Therefore, this study aimed to investigate the effect of fermented brown rice flour on white rice flour, white rice batter and its steamed bread qualities. Brown rice batter was fermented using commercial baker's yeast (Eagle brand) according to the optimum conditions for moderate acidity (pH 5.5) to obtain fermented brown rice flour (FBRF). The FBRF was added to white rice flour at 0%, 10%, 20%, 30%, 40% and 50% levels to prepare steamed rice bread. Based on the sensory evaluation test, steamed rice bread containing 40% FBRF had the highest overall acceptability score. Thus, pasting properties of the composite rice flour, rheological properties of its batter, volume and texture properties of its steamed bread were determined. The results showed that peak viscosity of the rice flour containing 40% FBRF was significantly increased, whereas its breakdown, final viscosity and setback significantly decreased. Viscous, elastic and complex moduli of the batter having 40% FBRF were also significantly reduced. However, volume, specific volume, chewiness, resilience and cohesiveness of its steamed bread were significantly increased, while hardness and springiness significantly reduced in comparison to the control. These results established the effectiveness of yeast fermentation in reducing the detrimental effects of bran on the sensory properties of steamed white rice bread and encourage the usage of brown rice flour to enhance the quality of rice products.
Kinetic analysis of solid-state fermentation (SSF) of fruit peels with Phanerochaete chrysosporium and Schizophyllum commune mixed culture was studied in flask and 7 kg capacity reactor. Modified Monod kinetic model suggested by Haldane sufficiently described microbial growth with co-efficient of determination (R2) reaching 0.908 at increased substrate concentration than the classical Monod model (R2 = 0.932). Leudeking-Piret model adequately described product synthesis in non-growth-dependent manner (R2 = 0.989), while substrate consumption by P. chrysosporium and S. commune fungal mixed culture was growth-dependent (R2 = 0.938). Hanes-Woolf model sufficiently represented α-amylase and cellulase enzymes synthesis (R2 = 0.911 and 0.988); α-amylase had enzyme maximum velocity (Vmax) of 25.19 IU/gds/day and rate constant (Km) of 11.55 IU/gds/day, while cellulase enzyme had Vmax of 3.05 IU/gds/day and Km of 57.47 IU/gds/day. Product yield in the reactor increased to 32.65 mg/g/day compared with 28.15 mg/g/day in shake flask. 2.5 cm media thickness was adequate for product formation within a 6 day SSF in the tray reactor.
This study aimed to enhance production of polyhydroxybutyrate P(3HB) by a newly engineered strain of Cupriavidus necator NSDG-GG by applying response surface methodology (RSM). From initial experiment of one-factor-at-a-time (OFAT), glucose and urea were found to be the most significant substrates as carbon and nitrogen sources, respectively, for the production of P(3HB). OFAT experiment results showed that the maximum biomass, P(3HB) content, and P(3HB) concentration of 8.95 g/L, 76 wt%, and 6.80 g/L were achieved at 25 g/L glucose and 0.54 g/L urea with an agitation rate of 200 rpm at 30 °C after 48 h. In this study, RSM was applied to optimize the three key variables (glucose concentration, urea concentration, and agitation speed) at a time to obtain optimal conditions in a multivariable system. Fermentation experiments were conducted in shaking flask by cultivation of C. necator NSDG-GG using various glucose concentrations (10-50 g/L), urea concentrations (0.27-0.73 g/L), and agitation speeds (150-250 rpm). The interaction between the variables studied was analyzed by ANOVA analysis. The RSM results indicated that the optimum cultivation conditions were 37.70 g/L glucose, 0.73 g/L urea, and 200 rpm agitation speed. The validation experiments under optimum conditions produced the highest biomass of 12.84 g/L, P(3HB) content of 92.16 wt%, and P(3HB) concentration of 11.83 g/L. RSM was found to be an efficient method in enhancing the production of biomass, P(3HB) content, and P(3HB) concentration by 43, 21, and 74%, respectively.
The increase in the price of commercial succinic acid has necessitated the need for its
synthesis from waste materials such as glycerol. Glycerol residue is a waste product of
Oleochemical production which is cheaply available and a very good source of carbon.
The use of immobilized cells can further reduce the overall cost of the production process.
This study primarily aims to produce succinic acid from glycerol residue through the use
of immobilized Escherichia coli in a batch fermentation process. The parameters which
affect bacterial fermentation process such as the mass substrate, temperature, inoculum
size and duration of fermentation were screened using One-Factor-At-a-Time (OFAT)
method. The result of the screening process shows that a substrate (glycerol) concentration
of 30 g, inoculum size 20% v/v, and time 4 h produced the maximum succinic acid
concentration of 117.99 g/L. The immobilized cells were found to be stable as well as
retain their fermentative ability up to the 6th cycle of recycling, thereby presenting as an
advantage over the free cell system. Therefore, conclude that using immobilized cells can
contribute immensely to the cost-effective production of succinic acid from glycerol
residue.
Lactobacillus plantarum is a non-gas-producing lactic acid bacterium that is generally regarded as safe (GRAS) with Qualified Presumption of Safety (QPS) status. Although traditionally used for dairy, meat and vegetable fermentation, L. plantarum is gaining increasing significance as a probiotic. With the newly acclaimed gut-heart-brain axis, strains of L. plantarum have proven to be a valuable species for the development of probiotics, with various beneficial effects on gut health, metabolic disorders and brain health. In this review, the classification and taxonomy, and the relation of these with safety aspects are introduced. Characteristics of L. plantarum to fulfill the criteria as a probiotic are discussed. Emphasis are also given to the beneficial functions of L. plantarum in gut disorders such as inflammatory bowel diseases, metabolic syndromes, dyslipidemia, hypercholesteromia, obesity, and diabetes, and brain health aspects involving psychological disorders.
Fermentation employing lactic acid bacteria (LAB) often suffers end-product inhibition which reduces the cell growth rate and the production of metabolite. The utility of adsorbent resins for in situ lactic acid removal to enhance the cultivation performance of probiotic, Pediococcus acidilactici was studied. Weak base anion-exchange resin, Amberlite IRA 67 gave the highest maximum uptake capacity of lactic acid based on Langmuir adsorption isotherm (0.996 g lactic acid/g wet resin) compared to the other tested anion-exchange resins (Amberlite IRA 410, Amberlite IRA 400, Duolite A7 and Bowex MSA). The application of Amberlite IRA 67 improved the growth of P. acidilactici about 67 times compared to the control fermentation without resin addition. Nevertheless, the in situ addition of dispersed resin in the culture created shear stress by resins collision and caused direct shear force to the cells. The growth of P. acidilactici in the integrated bioreactor-internal column system containing anion-exchange resin was further improved by 1.4 times over that obtained in the bioreactor containing dispersed resin. The improvement of the P. acidilactici growth indicated that extractive fermentation using solid phase is an effective approach for reducing by-product inhibition and increasing product titer.
MPT64 is a specific protein that is secreted by Mycobacterium tuberculosis complex (MTBC). The objective of this study was to obtain optimum culture conditions for MPT64 synthetic gene expression in Escherichia coli BL21 (DE3) by response surface methodology (RSM). The RSM was undertaken to optimize the culture conditions under different cultivation conditions (medium concentration, induction time and inducer concentration), designed by the factorial Box-Bhenken using Minitab 17 statistical software. From the randomized combination, 15 treatments and three center point repetitions were obtained. Furthermore, expression methods were carried out in the flask scale fermentation in accordance with the predetermined design. Then, the MPT64 protein in the cytoplasm of E. coli cell was isolated and characterized using sodium dodecyl sulfate polyacrilamide electrophoresis (SDS-PAGE) then quantified using the ImageJ program. The optimum conditions were two-fold medium concentration (tryptone 20 mg/mL, yeast extract 10 mg/mL, and sodium chloride 20 mg/mL), 5 h of induction time and 4 mM rhamnose. The average concentration of recombinant MPT64 at optimum conditions was 0.0392 mg/mL, higher than the predicted concentration of 0.0311 mg/mL. In conclusion, the relationship between the selected optimization parameters strongly influenced the level of MPT64 gene expression in E. coli BL21 (DE3).
The aim of this study is to identify the predominating lactic acid bacteria (LAB) in a spontaneous fermented wheat sourdough. At the same time, an investigation towards volatile compounds that were produced was also carried out. Lactobacillus plantarum has been identified as the dominant species of lactobacilli with characters of a facultative heterofermentative strain. The generated volatile compounds that were produced during spontaneous fermentation were isolated by solvent extraction method, analysed by gas chromatography (GC), and identified by mass spectrophotometer (MS). Butyric acid has been found to be the main volatile compound with relative abundance of 6.75% and acetic acid at relative abundance of 3.60%. Esters that were formed at relatively low amount were butyl formate (1.23%) and cis 3 hexenyl propionate (0.05%). Butanol was also found at low amount with relative abundance of 0.60%. The carbohydrate metabolism of Lactobacillus plantarum may contributed to the production of acetic acid in this study via further catabolism activity on lactic acid that was produced. However, butyric acid was not the major product via fermentation by LAB but mostly carried out by the genus Clostridium via carbohydrate metabolism which needs further investigation
The Cassava starch manufacturing wastewater (CSW) was used as a substrate to produce polyhydroxybutyrate (PHB) by Cupriavidus sp. KKU38. The acidogenic fermentation process of CSW was first conducted to obtain volatile fatty acids (VFAs), which are more efficient in PHB production than raw CSW. The effect on substrate concentration and nutrients, i.e. nitrogen and phosphorus concentrations, by means of chemical oxygen demand: nitrogen: phosphorus ratio (COD:N:P ratio) variation was investigated. The results indicated that PHB production from fermented CSW by Cupriavidus sp. KKU38 was optimized at the soluble COD:N:P ratio of 100:0.5:11. This ratio gave the maximum PHB content and yield of 85.53% and 0.31 g PHB/g COD consumed, respectively. By using the proposed PHB production process, the potential to produce 0.19 kg of PHB from 1.0 kg of soluble chemical oxygen demand (sCOD) contained in CSW was exhibited. The relatively high COD removal efficiency of 73.82% at the optimal condition could be achieved, which demonstrated the concept of water quality improvements alongside the production of the value-added by-product, PHB.
Mutagenic and antimutagenic activities of lactic acid bacteria (LAB) Lactobacillus plantarum isolated from the local fermented durian (tempoyak) was determined by Ames test (Salmonella/microsome mutagenicity assay). Our study also involved pre-incubation assay against Salmonella typhimurium TA 98 and TA 100 bacterial strain in the presence and absence of metabolic activator S9 system. It was found that the L. plantarum showed no mutagenic activity on both S. typhimurium strain TA 98 and TA 100 in the presence and absence of metabolic activator. Significant antimutagenic activity (p < 0.05) was observed in both cell-free supernatant and bacterial cell suspension of L. plantarum as compared to the mutagenicity induced by 2-Aminoanthracene in the presence of metabolic activator. Meanwhile, in the absence of metabolic activator, only the bacterial cells of L. plantarum showed antimutagenicity acitivity against Sodium Azide and 2-Nitrofluorene. In conclusion, L. plantarum could play a vital role as chemopreventive agent by binding to mutagens and suppressing mutagenesis. Thus, L. plantarum could be consider as a good candidate for functional food development as a supplement product to prevent development of colon cancer.
Bioethanol is a very environmentally friendly liquid biofuel that is not only renewable, but also sustainable. It is currently
deemed as a highly suitable additive and substitute energy source to replace fossil based fuel. In this study, bioethanol
was produced from sago hampas by using commercial amylase, cellulase and Saccharomyces cerevisiae via sequential
saccharification and simultaneous fermentation (SSSF), a modified version of the simultaneous saccharification and
fermentation (SSF) process. SSSF was performed on sago hampas at 2.5 and 5.0% (w/v) feedstock load for five days. The
samples taken from the SSSF broths were analysed via high performance liquid chromatography (HPLC) for ethanol, glucose
and acetic acid production. From the results obtained, SSSF with 5.0% sago hampas loading exhibited the highest ethanol
production at 14.13 g/L (77.43% of theoretical ethanol yield), while SSSF using 2.5% sago hampas loading produced
ethanol at 6.45 g/L (69.24% of theoretical ethanol yield). This study has shown that ethanol not only can be produced
from sago hampas using different enzyme mixtures and S. cerevisiae via SSSF, but yields were also high, making this
process highly promising for the production of cheap and sustainable ethanol as fuel.
Nipa (Nypa fruticans) is one of the most widely distributed and useful palm in the mangrove forests in the South, Southeast Asia and Oceania. Its distribution area is known to be larger in ancient time than at present, as evidenced by its fossils found in North America, South America, Egypt and Europe. Nipa has a wide diversity of use. Traditionally it is used as roof materials, cigarette wrapper, medicine and its sap is fermented to alcohol. Recently, research on nipa has focused on its potential use as a biofuel crop because it has several advantages compared with other biofuel-alcohol crops. For example it has high alcohol content, no competition with other crop for agricultural land and no bagasse disposal problem. In spite of such usefulness, scientific reports on biology of nipa are limited. Information on genetic diversity, cytogenetics and chemical composition are lacking for nipa plant. On the other hand, morphological characters of nipa have been described in many reports. This paper attempted to provide a general review of the nipa plant based on available literatures.
Sweatings, the exudates that leach out from fermenting fruits during rambutan fruit fermentation are considered as
a waste by-product and are allowed to be drained off. This could lead to a pollution problem. Besides, it is a waste if
the sweatings are possible to be transformed into food products and ingredients. However, prior transformation, the
fundamental knowledge of the sweatings should be understood. Hence, the main aim of this study was to investigate
the physicochemical properties of sweatings as affected by fermentation time and turning intervals during natural
fermentation of rambutan fruits. In this study, peeled rambutan fruit was fermented for 8 days and turned. Different
batches of the fruits were turned every 24, 48 or 72 h and sweatings from the process were collected and analyzed.
The results showed that fermentation time significantly reduced (p<0.05) the yield, pH and sucrose content of the
sweatings by 79-84%, 32-33%, 76.5-80.8%, respectively. Fermentation time also significantly increased (p<0.05) the
titratable acidity, total soluble solids, fructose, glucose, total sugar, citric acid, lactic acid, acetic acid and ascorbic
acid contents of the sweatings by 5.6-6.0, 1.5-1.6, 2.4-2.6, 2.1-2.5, 1.0-1.1, 5.7-6.5, 2.4-2.6, 2.1-2.5 and 2.6-2.8 folds,
respectively. However, turning intervals did not significantly affect (p>0.05) the physicochemical properties of the
sweatings. High concentrations of sugars and organic acids allow the sweatings to have a balance of sweet and sour
taste and they are suitable to be used in the production of syrup, soft drinks, jam, jelly, marmalade and vinegar.
Phytase activity and growth of anaerobic rumen bacterium, Mitsuokella jalaludinii were investigated by semi-solid
state fermentation. Carbon source (rice bran, yam and cassava), nitrogen sources (soya bean, offal meal, fish meal and
feather meal) and growth factors (hemin, L-cysteine hydrochloride and minerals) were evaluated in a one-factor-at-atime
approach. Rice bran and fish meal produced better growth and phytase enzyme activity. The removal of L-cysteine
hydrochloride and minerals significantly decreased (p<0.05) phytase activity from 1178.72 U to 446.99 U and 902.54
U, respectively. The response surface methods (RSM) was conducted to optimize the phytase production and the results
showed the combination of 7.7% of rice bran and 3.7% of fish meal in semi-solid state fermentation gave the highest
phytase activity. Maximum phytase production and optimum growth of bacteria were detected at 12 h incubation in both
MF medium (control) and agro-medium. In this agro-medium, M. jalaludinii produced 2.5 fold higher phytase activity
compared to MF medium.