Continuous fermentation of dilute acid-pretreated de-oiled rice bran (DRB) to butanol by the Clostridium acetobutylicum YM1 strain was investigated. Pretreatment of DRB with dilute sulfuric acid (1%) resulted in the production of 42.12 g/L total sugars, including 25.57 g/L glucose, 15.1 g/L xylose and 1.46 g/L cellobiose. Pretreated-DRB (SADRB) was used as a fermentation medium at various dilution rates, and a dilution rate of 0.02 h-1 was optimal for solvent production, in which 11.18 g/L of total solvent was produced (acetone 4.37 g/L, butanol 5.89 g/L and ethanol 0.92 g/L). Detoxification of SADRB with activated charcoal resulted in the high removal of fermentation inhibitory compounds. Fermentation of detoxified-SADRB in continuous fermentation with a dilution rate of 0.02 h-1 achieved higher concentrations of solvent (12.42 g/L) and butanol (6.87 g/L), respectively, with a solvent productivity of 0.248 g/L.h. This study showed that the solvent concentration and productivity in continuous fermentation from SADRB was higher than that obtained from batch culture fermentation. This study also provides an economic assessment for butanol production in continuous fermentation process from DRB to validate the commercial viability of this process.
Postbiotic RS5, produced by Lactiplantibacillus plantarum RS5, has been identified as a promising alternative feed supplement for various livestock. This study aimed to lower the production cost by enhancing the antimicrobial activity of the postbiotic RS5 by improving the culture density of L. plantarum RS5 and reducing the cost of growth medium. A combination of conventional and statistical-based approaches (Fractional Factorial Design and Central Composite Design of Response Surface Methodology) was employed to develop a refined medium for the enhancement of the antimicrobial activity of postbiotic RS5. A refined medium containing 20 g/L of glucose, 27.84 g/L of yeast extract, 5.75 g/L of sodium acetate, 1.12 g/L of Tween 80 and 0.05 g/L of manganese sulphate enhanced the antimicrobial activity of postbiotic RS5 by 108%. The cost of the production medium was reduced by 85% as compared to the commercially available de Man, Rogosa and Sharpe medium that is typically used for Lactobacillus cultivation. Hence, the refined medium has made the postbiotic RS5 more feasible and cost-effective to be adopted as a feed supplement for various livestock industries.
The aim of this study was to establish an improved pretreatment and fermentation method i.e. immobilized cells for high recovery of fermentable sugars from palm kernel cake (PKC) and its effects on fermentability performance by Actinobacillus succinogenes 130Z in the conversion of the fermentable sugar to lactic acid. The effects of oxalic acid concentrations (1-6% w/v) and residence times (1-5 h) on the sugar recovery were initially investigated and it was found that the highest mannose concentration was 25.1 g/L at the optimum hydrolysis conditions of 4 h and 3% (w/v) oxalic acid. The subsequent enzymatic saccharification of the pretreated PKC afforded the highest enzymatic digestibility with the recovered sugars amounting to 25.18 g/L and 9.14 g/L of mannose and glucose, respectively. Subsequently, the fermentability performance of PKC hydrolysate was evaluated and compared in terms of cultivation phases (i.e. mono and dual-phases), carbonate loadings (i.e. magnesium and sodium carbonates), and types of sugars (i.e. glucose and mannose). The highest titer of 19.4 g/L lactic acid was obtained from the fermentation involving A. succinogenes 130Z in dual-phase cultivation supplemented with 30 g/L of magnesium carbonate. Lactic acid production was further enhanced by using immobilized cells with coconut shell-activated carbon (CSAC) of different sizes (A, B, C, and D) in the repeated batch cultivation of dual-phase fermentation producing 31.64 g/L of lactic acid. This work sheds light on the possibilities to enhance the utilization of PKC for lactic acid production via immobilized A. succinogenes 130Z.
This study explores utilizing pineapple peel (PP) hydrolysate as a promising carbon source for xylitol production, covering scopes from the pre-treatment to the fermentation process. The highest xylose concentration achieved was around 20 g/L via mild acid hydrolysis (5% nitric acid, 105 °C, 20-min residence time) with a solid loading of 10%. Two sets fermentability experiments were carried out of varying pH levels in synthetic media that includes acetic acid as the main inhibitors and hydrolysate supplemented with diverse nitrogen source. The results revealed that pH 7 exhibited the highest xylitol production, yielding 0.35 g/g. Furthermore, urea was found to be a highly promising and cost-effective substitute for yeast extract, as it yielded a comparable xylitol production of 0.31 g/g with marginal difference of only 0.01 g/g compared to yeast extract further highlights the viability of urea as the preferred option for reducing xylitol production cost. The absence of a significant difference between the synthetic media and hydrolysate, with only a marginal variance of 0.35 to 0.32 g/g, implies that acetic acid is indeed the primary constraint in xylitol production using PP hydrolysate. The study sheds light on PP biomass's potential for xylitol production, aligning economic benefits with environmental sustainability and waste management.
Fungi is known to produce a wide range of biologically active metabolites and enzymes. Enzymes produced by fungi are utilized in food and pharmaceutical industries because of their rich enzymatic profile. Filamentous fungi are particularly interesting due to their high production of extracellular enzymes which has a large industrial potential. The aim of this study is to isolate potential soil fungi species that are able to produce functional enzymes for industries. Five Aspergillus species were successfully isolated from antibiotic overexposed soil (GPS coordinate of N3.093219 E101.40269) by standard microbiological method. The isolated fungi were identified via morphological observations and molecular tools; polymerase chain reactions, ITS 1 (5’- TCC GTA GGT GAA CCT GCG G3’) forward primer and ITS 4 (5’-TCC TCC GCT TAT TGA TAT GC-3’) reverse primer. The isolated fungi were identified as Aspergillus sydowii strain SCAU066, Aspergillus tamarii isolate TN-7, Aspergillus candidus strain KUFA 0062, Aspergillus versicolor isolate BAB-6580, and Aspergillus protuberus strain KAS 6024. Supernatant obtained via submerged fermentation of the isolated fungi in potato dextrose broth (PDB) and extracted via centrifugation was loaded onto specific media to screen for the production of xylanolytic, cellulolytic and amylolytic enzymes. The present findings indicate that Aspergillus sydowii strain SCAU066 and Aspergillus versicolor isolate BAB-6580 have great potential as an alternative source of xylanolytic, cellulolytic and amylolytic enzymes.
This research was performed based on a comparative study on fungal lipid production by a locally isolated strain Cunninghamella bainieri 2A1 in batch culture and repeated-batch culture using a nitrogen-limited medium. Lipid production in the batch culture was conducted to study the effect of different agitation rates on the simultaneous consumption of ammonium tartrate and glucose sources. Lipid production in the repeated-batch culture was studied by considering the effect of harvesting time and harvesting volume of the culture broth on the lipid accumulation. The batch cultivation was carried out in a 500 ml Erlenmeyer flask containing 200 ml of the fresh nitrogen-limited medium. Microbial culture was incubated at 30 °C under different agitation rates of 120, 180 and 250 rpm for 120 h. The repeated-batch culture was performed at three harvesting times of 12, 24 and 48 h using four harvesting cultures of 60%, 70%, 80% and 90%. Experimental results revealed that nitrogen source (ammonium tartrate) was fully utilized by C. bainieri 2A1 within 24 h in all agitation rates tested. It was also observed that a high amount of glucose in culture medium was consumed by C. bainieri 2A1 at 250 rpm agitation speed during the batch fermentation. Similar results showed that the highest lipid concentration of 2.96 g/L was obtained at an agitation rate of 250 rpm at 120 h cultivation time with the maximum lipid productivity of 7.0 × 10(-2) mg/ml/h. On the other hand, experimental results showed that the highest lipid concentration produced in the repeated-batch culture was 3.30 g/L at the first cycle of 48 h harvesting time using 70% harvesting volume, while 0.23 g/L gamma-linolenic acid (GLA) was produced at the last cycle of 48 h harvesting time using 80% harvesting volume.
Food waste (FW) minimization at the source by using food waste biodigester (FWBs) has a vast potential to lower down the impact of increasing organic fraction in municipal solid waste generation. To this end, this research sought to check the performance of locally isolated hydrolase-producing bacteria (HPB) to improve food waste biodegradation rate. Two under-explored HPB identified as Bacillus paralicheniformis GRA2 and Bacillus velezensis TAP5 were able to produce maximum amylase, cellulase, protease and lipase activities, and demonstrated a significant hydrolase synergy in co-culture fermentation. In vitro biodegradation analysis of both autoclaved and non-autoclaved FW revealed that the HPB inoculation was effective to degrade total solids (>62%), protein (>19%), total fat (>51), total sugar (>86%), reducing sugar (>38%) and starch (>50%) after 8-day incubation. All co-culture treatments were recorded superior to the respective monocultures and the uninoculated control. The results of FW biodegradation using batch-biodigester trial indicated that the 1500 mL and 1000 mL inoculum size of HPB inoculant reached a plateau on the 4th day, with gross biodegradation percentage (GBP) of >85% as compared to control (66.4%). The 1000 mL inoculum was sufficient to achieve the maximum GBP (>90%) of FW after an 8-day biodigestion in a FWB.
The effect of pH and butyric acid supplementation on the production of butanol by a new local isolate of Clostridium acetobutylicum YM1 during batch culture fermentation was investigated. The results showed that pH had a significant effect on bacterial growth and butanol yield and productivity. The optimal initial pH that maximized butanol production was pH 6.0 ± 0.2. Controlled pH was found to be unsuitable for butanol production in strain YM1, while the uncontrolled pH condition with an initial pH of 6.0 ± 0.2 was suitable for bacterial growth, butanol yield and productivity. The maximum butanol concentration of 13.5 ± 1.42 g/L was obtained from cultures grown under the uncontrolled pH condition, resulting in a butanol yield (YP/ S ) and productivity of 0.27 g/g and 0.188 g/L h, respectively. Supplementation of the pH-controlled cultures with 4.0 g/L butyric acid did not improve butanol production; however, supplementation of the uncontrolled pH cultures resulted in high butanol concentrations, yield and productivity (16.50 ± 0.8 g/L, 0.345 g/g and 0.163 g/L h, respectively). pH influenced the activity of NADH-dependent butanol dehydrogenase, with the highest activity obtained under the uncontrolled pH condition. This study revealed that pH is a very important factor in butanol fermentation by C. acetobutylicum YM1.
This study examined lactic acid bacteria (LAB)-fermented soymilk for their ability in hydrolyzing glucosides to aglycones
and corresponding antioxidant capacity and memory enhancing effect. Twelve LAB isolated from Malaysian fermented food
and milk products were incubated in commercially available soymilk for 48 h. Generally, soymilk supported LAB growth
and significantly increased (p<0.05) conversion to bioactive aglycone by 2.1-6.5 fold when compared to unfermented
soymilk. Lactobacillus fermentum LAB 9- fermented soymilk, in particular, was presented with increased total phenolic
content (+10%) as opposed to unfermented soymilk. Lactobacilli (LAB 10-12)- and pediococci (LAB 5)-fermented soymilk
elicited maximal DPPH radical-scavenging activity. LAB 1, 7, 8, 9 and 12 exhibited significantly higher (p<0.05) ferrous
ion chelating activity when compared to control. Interestingly, LAB 9 had significantly improved memory deficit (p<0.05)
in LPS-challenged mice. LAB-enriched nutritional value of soymilk could be useful against oxidative stress and memory
deficit.
Bacterial adhesion to host cells is the most important probiotic character. However, the adhesion of probiotic should not
affect the viability of the host cells. In this study, Lactobacillus plantarum strain L8, Lactobacillus plantarum strain L20
and Lactobacillus pentosus strain S1 were tested for their cytotoxic effects through MTT assay and their ability to adhere
and colonize on HT-29 and CCD-18Co intestinal cells as detected microscopically using light microscopy and Scanning
Electron Microscopy (SEM). No cytotoxicity effects were observed on both intestinal cells following 24 h treatment with
all Lactobacillus strains. Additionally, all strains demonstrated strong adhesive activity where more than 100 bacteria
adhered to both intestinal cells although differences in the adhesion scores observed among different strains. The adhesion
as observed via SEM showed an autoagreggative pattern and adhered as clusters on the surface of both intestinal cells.
In conclusion, all three Lactobacillus strains are non-cytotoxic to both cells with strong adhesion ability on intestinal
cells and this study also proved that Malaysian fermented fish are good source of probiotic bacteria.
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 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.
Acetobacter xylinum strains are known as efficient producers of cellulose. A. xylinum is an obligate aerobic bacterium that has an oxygen-based metabolism. The dissolved oxygen (DO) concentration in a rotary discs reactor (RDR) is one of the most important factors that need to be observed during the cellulose synthesis by these bacteria. In this study, the effects of different discs rotation speed (5, 7, 9 and 12 rpm) and fermentation period (3, 4, 5 and 6 days) on the DO concentration and production of bacterial cellulose in a 10-L RDR were examined. The highest yield was obtained at 7 rpm with a total dried weight of 28.3 g for 4 days fermentation. The results showed that the DO concentration in the 10-L RDR increased in the range of 13 to 17% with increasing of discs rotation speed from 7 to 12 rpm. However, fermentation with high discs rotation speed at 12 rpm reduced the bacterial cellulose production. Analysis of data using Statistica 8.0 showed a high coefficient of determination value (R2 = 0.92). In conclusion, discs rotation speed gave more significant effect on the DO concentration and production of bacterial cellulose in 10-L RDR compared to fermentation period. This was further combined with synergistic effect from sufficient consumption of oxygen for the enhanced production of bacterial cellulose and providing the controlled environment for encouraging bacterial growth throughout the fermentation process.
Ethanol fermentations by Candida shehatae TISTR 5843 at low (20 g/L) and high (80 g/L) sugar concentrations with various glucose to xylose ratios were investigated. Glucose was a preferred substrate as it was consumed first at a faster consumption rate. The type of sugar and ratio between glucose and xylose did not have an effect on ethanol produced. The average ethanol concentrations were 7.99 g/L when using 20 g/L sugar and 27.82 g/L when using 80 g/L sugar. Small amounts of xylitol and glycerol as by-products were presented when using 20 g/L sugar. Xylitol appeared to be the main by-product at high xylose concentration with elevated concentrations as xylose is increased. When using rice straw hydrolysate containing 34.75 g/L glucose and 21.29 g/L xylose, 19.37 g/L ethanol was produced with the ethanol yield and ethanol productivity at 0.49 g/g and 0.20 g/L.h, respectively. However, xylose was not completely consumed after fermentation was complete.
During fermentation cells are subjected to various kinds of stress. One of the stresses concerned is high osmotic environment, which cells need to encounter in order to continue growing. To understand how cells adapt to this stress condition, information from genome, proteome and metabolome levels are crucial. In yeast cells, it was report that they produce glycerol to avoid depletion of water in the cell that could lead to cell shrinkage and eventually death. Thus, investigation of physiological responses were executed by shake flask method using three different Saccharomyces cerevisiae strains namely s288c, IFO2347 and FY834 which were grown in yeast potato dextrose (YPD) medium under the treatment of sodium chloride (NaCl) and sorbitol at 1M concentration to create the osmotic condition. These agents were added into the medium after 5 hours of fermentation when the cells reached exponential phase and carbon source is still available. The results proved that addition of both NaCl and sorbitol created the osmotic condition during growth resulted in higher accumulation of glycerol and trehalose when compared to the control in all strains. Among these strains, production of glycerol (g glycerol/g cell dry weight) was found highest in IFO2347, followed by s288c and FY834.
The application of solar disinfection for treating stored rainwater was investigated by the authors using indicator organisms. The multiple tube fermentation technique and pour plate method were used for the detection of microbial quality indicators like total and fecal coliforms, E. coli and heterotrophic plate count. These techniques have disadvantages mainly that these are laborious and time consuming. The correlation of total coliform with that of exposure time is proposed under different factors of weather, pH and turbidity. Statistical tools like root mean square error and coefficient of determination were used to validate these proposed equations. The correlation equations of fecal coliform, E. coli and heterotrophic plate count with total coliform are suggested by using four regression analysis including Reciprocal Quadratic, Polynomial Regression (2 degree), Gaussian Model and Linear Regression in order to reduce the tedious experimental work in similar types of experiments and treatment systems.
Palm oil mill effluent (POME) can be utilised directly as the sole substrate in the anaerobic fermentation of acetone-butanol-ethanol (ABE) and hydrogen by Clostridium acetobutylicum NClMB13357 in a submerged batch system. Effects of sedimented POME concentration and the initial culture pH on the production of ABE/H were studied. Sedimented POME with 90% v/v (POME90) at pH 5.8 is capable of producing 4.01 g/L ABE with acetone concentration at 1.97 g/L; butanol 1.74 g/L and ethanol 0.3 g/L. The highest concentration of butanol (1.86 g/L) was produced from a culture with initial pH 6.0. The production of hydrogen gas was proportioned to the concentration of POME. The highest hydrogen gas production was at pH 5.5 (31 mL). More than 50% (v/v) of hydrogen gas was produced at different pH except pH 4.5, when only 16% (v/v) or 5 mL of hydrogen was produced.
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.
Biological fermentation of Rhizopus oryzae was introduced to extract cellulose nanofibre from durian skin fibre (DSF).
The diameter of the extracted durian skin nanofibre (DSNF) was in the range of 49-81 nm. The changes of chemical
composition of DSNF were clearly seen after evaluated via TAPPI standard test methods. Verification via Fourier transform
infrared (FTIR) confirmed the deduction of hemicelluloses and lignin in DSNF in the range of 1200 to 1000 cm-1. X-ray
diffraction (XRD) demonstrated increment in the crystallinity from 58.3 to 72.2% after biological fermentation. DSNF was
then incorporated into polylactic acid (PLA) via extrusion and injection moulding processes. The effect of 1-5 wt. % DSNF
content on PLA biocomposites was investigated for its mechanical and thermal properties. The presence of only 1 wt. %
improved the tensile and impact strength by 14.1 MPa and 33.1 kJ/m2
, respectively. The thermal properties of PLA-1DSNF
biocomposite also recorded higher thermal stability, glass transition temperature (Tg
), crystallization temperature (Tc
)
and melting temperature (Tm). Additionally, from the DMA, it was determined that PLA-1DSNF possessed lower storage
modulus and loss modulus, as well as low energy dissipation.