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.
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 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.
Several members of the genus Lignosus, which are collectively known as cendawan susu rimau (in Malay) or tiger׳s milk mushrooms (TMM), are regarded as important local medicine particularly by the indigenous communities in Malaysia. The mushroom sclerotia are purportedly effective in treating cancer, coughs, asthma, fever, and other ailments. The most commonly encountered Lignosus spp. in Malaysia was authenticated as Lignosus rhinocerotis (Cooke) Ryvarden (synonym: Polyporus rhinocerus), which is also known as hurulingzhi in China and has been used by Chinese physicians to treat liver cancer, gastric ulcers, and chronic hepatitis. In spite of growing interest in the therapeutic potential of TMM, there is no compilation of scientific evidence that supports the ethnomedicinal uses of these mushrooms. Therefore, the present review is intended (i) to provide a comprehensive, up-to-date overview of the ethnomedicinal uses, pharmacological activities, and cultivation of TMM in general and L. rhinocerotis in particular, (ii) to demonstrate how recent scientific findings have validated some of their traditional uses, and (iii) to identify opportunities for future research and areas to prioritize for TMM bioprospecting.
The fermentation of cocoa pulp is one of the few remaining large-scale spontaneous microbial processes in today's food industry. The microbiota involved in cocoa pulp fermentations is complex and variable, which leads to inconsistent production efficiency and cocoa quality. Despite intensive research in the field, a detailed and comprehensive analysis of the microbiota is still lacking, especially for the expanding Asian production region. Here, we report a large-scale, comprehensive analysis of four spontaneous Malaysian cocoa pulp fermentations across two time points in the harvest season and two fermentation methods. Our results show that the cocoa microbiota consists of a "core" and a "variable" part. The bacterial populations show a remarkable consistency, with only two dominant species, Lactobacillus fermentum and Acetobacter pasteurianus. The fungal diversity is much larger, with four dominant species occurring in all fermentations ("core" yeasts), and a large number of yeasts that only occur in lower numbers and specific fermentations ("variable" yeasts). Despite this diversity, a clear pattern emerges, with early dominance of apiculate yeasts and late dominance of Saccharomyces cerevisiae. Our results provide new insights into the microbial diversity in Malaysian cocoa pulp fermentations and pave the way for the selection of starter cultures to increase efficiency and consistency.
Biobased L-lactic acid (L-LA) appeals to industries; however, existing technologies are plagued by limited productivity and high energy consumption. This study established an integrated process for producing macroalgae-based L-LA from Eucheuma denticulatum phycocolloid (EDP). Dilute acid-assisted microbubbles-mediated ozonolysis (DAMMO) was selected for the ozonolysis of EDP to optimize D-galactose recovery. Through single-factor optimization of DAMMO treatment, a maximum D-galactose recovery efficiency (59.10 %) was achieved using 0.15 M H2SO4 at 80 °C for 75 min. Fermentation with 3 % (w/v) mixed microbial cells (Bacillus coagulans ATCC 7050 and Lactobacillus acidophilus-14) and fermented residues achieved a 97.67 % L-LA yield. Additionally, this culture approach was further evaluated in repeated-batch fermentation and showed an average L-LA yield of 93.30 %, providing a feasible concept for macroalgae-based L-LA production.
This study investigated the effects of pineapple peel powder with varied chemical profiles and sonication-assisted polyphenol biotransformation during fermentation on the quality characteristics of yogurt products. It aimed at exploring the feasibility of sonication-assisted fermentation to enhance the physicochemical properties, control post-acidification, and improve antioxidant activities in yogurts fortified with polyphenol-rich pineapple peel powder. Targeted analysis showed that polyphenol-rich pineapple dietary fiber obtained by ultrasonication-assisted extraction (NPFU) exhibited the slowest rates of acidification, highest antioxidant capacity, and lowest degree of whey separation at 21.67 %. Sonication pretreatments significantly increased transformation of free phenolic acids derived from pineapple peel fiber during fermentation, particularly increasing the accumulation of ferulic acid, caffeic acid and 5-hydroxyflavone, revealing the positive effects of sonication-mediated fermentation in promoting the hydrolysis of conjugated phenolics into free fractions. Yogurts fortified with pineapple peel fiber displayed significantly higher antioxidant activities (p
Lignocellulosic biomass is a valuable, renewable substrate for the synthesis of polyhydroxybutyrate (PHB), an ecofriendly biopolymer. In this study, bacterial strain E5-3 was isolated from soil in Japan; it was identified as Burkholderia ambifaria strain E5-3 by 16 S rRNA gene sequencing. The strain showed optimal growth at 37 °C with an initial pH of 9. It demonstrated diverse metabolic ability, processing a broad range of carbon substrates, including xylose, glucose, sucrose, glycerol, cellobiose, and, notably, palm oil. Palm oil induced the highest cellular growth, with a PHB content of 65% wt. The strain exhibited inherent tolerance to potential fermentation inhibitors derived from lignocellulosic hydrolysate, withstanding 3 g/L 5-hydroxymethylfurfural and 1.25 g/L acetic acid. Employing a fed-batch fermentation strategy with a combination of glucose, xylose, and cellobiose resulted in PHB production 2.7-times that in traditional batch fermentation. The use of oil palm trunk hydrolysate, without inhibitor pretreatment, in a fed-batch fermentation setup led to significant cell growth with a PHB content of 45% wt, equivalent to 10 g/L. The physicochemical attributes of xylose-derived PHB produced by strain E5-3 included a molecular weight of 722 kDa, a number-average molecular weight of 191 kDa, and a polydispersity index of 3.78. The amorphous structure of this PHB displayed a glass transition temperature of 4.59 °C, while its crystalline counterpart had a melting point of 171.03 °C. This research highlights the potential of lignocellulosic feedstocks, especially oil palm trunk hydrolysate, for PHB production through fed-batch fermentation by B. ambifaria strain E5-3, which has high inhibitor tolerance.
Istamycins (ISMs) are 2-deoxyfortamine-containing aminoglycoside antibiotics (AGAs) produced by Streptomyces tenjimariensis ATCC 31603 with broad-spectrum bactericidal activities against most of the clinically relevant pathogens. Therefore, this study aimed to statistically optimize the environmental conditions affecting ISMs production using the central composite design (CCD). Both the effect of culture media composition and incubation time and agitation rate were studied as one factor at the time (OFAT). The results showed that both the aminoglycoside production medium and the protoplast regeneration medium gave the highest specific productivity. Results also showed that 6 days incubation time and 200 rpm agitation were optimum for their production. A CCD quadratic model of 17 runs was employed to test three key variables: initial pH, incubation temperature, and concentration of calcium carbonate. A significant statistical model was obtained including, an initial pH of 6.38, incubation temperature of 30 ˚C, and 5.3% CaCO3 concentration. This model was verified experimentally in the lab and resulted in a 31-fold increase as compared to the unoptimized conditions and a threefold increase to that generated by using the optimized culture media. To our knowledge, this is the first report about studying environmental conditions affecting ISM production as OFAT and through CCD design of the response surface methodology (RSM) employed for statistical optimization. In conclusion, the CCD design is an effective tool for optimizing ISMs at the shake flask level. However, the optimized conditions generated using the CCD model in this study should be scaled up in a fermenter for industrial production of ISMs by S. tenjimariensis ATCC 31603 considering the studied environmental conditions that significantly influence the production proces.
Whole-cell immobilisation technology involving ℽ-aminobutyric acid GABA biosynthesis using lactic acid bacteria (LAB) has been extensively studied owing to its numerous benefits over free-living bacteria, including enhanced productivity, improved cell viability, ability to prevent cell lysis and protect cells against bacteriophages and other stressful conditions. Therefore, a novel LAB biocatalyst was developed using various fruit and fruit waste, immobilising a potential probiotic strain, Lactiplantibacillus plantarum B7, via an adsorption method to improve GABA and cell viability. Apple and watermelon rind have been known to be the ideal natural supports for L. plantarum B7 owing to higher GABA and lactic acid production and improved cell viability among the other natural supports tested and selected to be used in repeated batch fermentation (RBF) to improve GABA production and cell viability. In general, immobilisation of L. plantarum B7 on natural support has better GABA and lactic acid production with improved cell viability via RBF compared to free cells. Watermelon rind-supported cells and apple-supported cells could produce nine and eight successful GABA cycles, respectively, within RBF, whereas free cells could only produce up to four cycles. When using watermelon rind-supported cells and apple-supported cells in RBF, the GABA titer may be raised by up to 6.7 (218.480 ± 0.280 g/L) and 6 (195.439 ± 0.042 g/L) times, respectively, in comparison to GABA synthesis by free cells in single batch fermentation (32.65 ± 0.029 g/L). Additionally, natural support immobilised L. plantarum B7 could retain half of its cell viability even after the 12th cycle of RBF, while no cell was observed in control.
The primary biological treatment method for organic sludge is composting and/or anaerobic digestion, but their product (compost or biogas) is of little economic benefit; therefore, an improved process to produce a high-value product is required to make sludge management more sustainable. Maximizing NH3 gas recovery during composting processes has the potential benefit of producing high-value microalgal biomass. However, the majority of produced ammonia does not evaporate as NH3 gas but retains as NH4+-N in the compost after fermentation. The present study investigates the effects of the timing of Ca(OH)2 dosing (on days 2, 5, and 9), and the Ca(OH)2 dose (1.1-2.6 mmol/batch), on lab-scale thermophilic composting of anaerobic sludge. The effects on NH3 recovery, organic matter degradability, and microbial activity are evaluated. Ca(OH)2 dosing immediately improved the emission of NH3, with yields 50-69% higher than those under control conditions. The timing of the dosing did not influence NH3 recovery or organic matter degradability. Higher Ca(OH)2 doses resulted in higher NH3 recovery, while microbial activity was temporarily and marginally inhibited. The pH of the compost reached 10-11.5 but quickly dropped to 8-8.5 within a day, probably because of neutralization of Ca(OH)2 by the emitted CO2 and release of NH3, which maintained the microbial activity. The present study indicated that Ca(OH)2 dosing would be useful to apply during thermophilic composting for NH3 recovery to cultivate high-value microalgal biomass, which enables this process to obtain a more economic benefit.
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.
Response surface methodology (RSM) with central composite design (CCD) was applied to optimize key factors affecting
hydrogen production (HP) from diluted acid hydrolysate of water-hyacinth stem (WHS) by heat-treated anaerobic sludge
in a batch fermentation process. Key factors affecting namely substrate concentration and initial pH was investigated.
The results indicated that substrate concentration and initial pH had significantly effects on HP (p<0.05). A maximum HP
hydrogen production rate and hydrogen yield of 182.7 mmol H2
/L, 2.81 mmol H2
/L h and 0.84 mol H2
/mol hexose were
obtained under the optimum conditions i.e. substrate concentration of 4.06 g/L and initial pH of 5.81. The total energy
production from the fermentative of WHS hydrolysate was 1.97 kJ.
FERMSOSTAT is a developed laboratory scale solid state fermenter. It is a horizontal stirrer drum bioreactor with about 70 L capacities. The fermenter is made of stainless steel which is anti-corrosive and non-toxic to the process organism. The fermenter is equipped with sets of control systems for temperature, agitation, aeration and also outlets for substrate sampling as well as inlets for inoculation and substrate additions. The uniqueness of this FERMSOSTAT system is its ability to carry out in situ substrate sterilization and extraction of enzymes at the end of SSF process. Moreover, the mixing system provided by FERMSOSTAT can be performed either full or half mixing as well as forward or reverse mixing. Furthermore, the mixing can be programmed to run at certain agitation rate and time interval during the fermentation process to prevent or reduce damage to the fungus mycelia. FERMSOSTAT is a developed SSF bioreactor and not an improvement of any existing one. The performances of FERMSOSTAT have been evaluated. Under optimum solid state fermentation conditions, about 63.4, 397 and 3.21 U/g of CMCase, xylanase and FPase activities were detected, which were higher compared to the tray system.
An innovative approach was developed by incorporating high-pressure CO2 into the separate hydrolysis-fermentation of aspen leftover branches, aiming to enhance the bioethanol production efficiency. The high-pressure CO2 significantly increased the 72-h enzymatic hydrolysis yield of converting aspen into glucose from 53.8% to 82.9%. The hydrolysis process was performed with low enzyme loading (10 FPU g-1 glucan) with the aim of reducing the cost of fuel bioethanol production. The ethanol yield from fermentation of the hydrolyzed glucose using yeast (Saccharomyces cerevisiae) was 8.7 g L-1, showing increment of 10% compared with the glucose control. Techno-economic analysis indicated that the energy consumption of fuel bioethanol production from aspen branch chips was reduced by 35% and the production cost was cut 44% to 0.615 USD L-1, when 68 atm CO2 was introduced into the process. These results furtherly emphasized the low carbon footprint of this sustainable energy production approach.
This study highlights biohydrogen production enrichment through NiO and CoO nanoparticles (NPs) inclusion to dark fermentation of rice mill wastewater using Clostridium beijerinckii DSM 791. NiO (~26 nm) and CoO (~50 nm) NPs were intrinsically prepared via facile hydrothermal method with polyhedral morphology and high purity. Dosage dependency studies revealed the maximum biohydrogen production characteristics for 1.5 mg/L concentration of both NPs. Biohydrogen yield was improved by 2.09 and 1.9 folds higher for optimum dosage of NiO and CoO respectively, compared to control run without NPs. Co-metabolites analysis confirmed the biohydrogen production through acetate and butyrate pathways. Maximum COD reduction efficiencies of 77.6% and 69.5% were observed for NiO and CoO inclusions respectively, which were higher than control run (57.5%). Gompertz kinetic model fitted well with experimental data of NPs assisted fermentation. Thus, NiO and CoO inclusions to wastewater fermentation seems to be a promising technique for augmented biohydrogen production.
To set a benchmark in fungal growth rate, a differential analysis of prototrophic Aspergillus fumigatus AR04 with three ascomycetes applied in > 103 t year-1 scale was performed, i.e. Ashbya gosspyii (riboflavin), Aspergillus niger (citric acid) and Aspergillus oryzae (food-processing). While radial colony growth decreased 0.5-fold when A. gossypii was cultivated at 40°C instead of 28°C, A. fumigatus AR04 responded with 1.7-fold faster hyphal growth. A. niger and A. oryzae formed colonies at 40°C, but not at 43°C. Moreover, all A. fumigatus strains tested grew even at 49°C. In chemostat experiments, A. fumigatus AR04 reached steady state at a dilution rate of 0.7 h-1 at 40°C, 120% more than reported for A. gossypii at 28°C. To study mycelial growth rates under unlimited conditions, carbon dioxide increase rates were calculated from concentrations detected online in the exhaust of batch fermentations for 3 h only. All rates calculated suggest that A. fumigatus AR04 approximates Arrhenius' rule when comparing short cultivations at 30°C with those at 40°C. Linearization of the exponential phase and comparison of the slopes revealed an increase to 192% by the 10°C up-shift.
Response Surface Methodology (RSM) was used in the study to optimize the production of vanillin from isoeugenol through fermentation by Aspergillus niger I-1472. Three factors were studied which include amount of isoeugenol, resin (Amberlite XAD-4) and Span 80. During fermentation, isoeugenol as substrate were vortexed with Span 80 and added into the culture on Day 4. Resin (Amberlite XAD-4) was added into the medium the following day. The predicted optimum medium combination consisted of 3.61 g/L of isoeugenol, 5.8% (g/ mL) of Amberlite XAD-4 resin and 0.37% of Span 80 with an expected vanillin production of 0.137 g/L. Verification test showed that the model produced similar predicted and experimental values.
Two sequential statistical experimental designs were used to screen and investigate the dependence of the amount of biodegradation of Procion Red MX-8B (PR-MX8B) on the fermentation variables. Fourteen factors were screened using the Plackett-Burman design. Among these factors, the most significant variables which included yeast extract, corn steep solids and starch influencing PR-MX8B decolourisation were statistically elucidated for optimization. The optimum concentrations of 5.00 g/l yeast extract, 2.99 g/l starch and 1.89 g/l corn steep solids were predicted by applying the Box-Behnken design to the second order polynomial model fitted to the results obtained. The best predicted optimal conditions verified experimentally yielded 72.11% while the predicted value from the polynomial model was 79.17%. The experimental values were in good agreement with the predicted values with a 90.81% degree of accuracy.
Biocellulose (BC), produced by Acetobacter xylinum (0416), was carried out using three types of medium
composition under static surface culture. The media used in this experiment included CWHSM (Coconut
water in Hestrin-Schramm medium), CM (Complex medium) and HSM (Hestrin-Schramm medium).
CWHSM and CM used coconut water from agro-waste as the main source of sugar. The fermentation
was conducted for 12 days and the results of BC dry weight, cell entrapped, pH medium and productivity
were evaluated and compared. The results show that CWHSM is the most suitable medium for BC
production with a productivity of up to 0.044 g l
-1
day
-1
.