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.
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.
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.
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.
In this work, fractionation of empty fruit bunch (EFB) by bisulfite pretreatment was studied for the production of bioethanol and high value products to achieve biorefinery of EFB. EFB was fractionated to solid and liquor components by bisulfite process. The solid components were used for bioethanol production by quasi-simultaneous saccharification and fermentation. The liquor components were then converted to furfural by hydrolysis with sulfuric acid. Preliminary results showed that the concentration of furfural was highest at 18.8g/L with 0.75% sulfuric acid and reaction time of 25min. The conversion of xylose to furfural was 82.5%. Furthermore, we attempted to fractionate the liquor into hemicellulose sugars and lignin by different methods for producing potential chemicals, such as xylose, xylooligosaccharide, and lignosulfonate. Our research showed that the combination of bisulfite pretreatment and resin separation could effectively fractionate EFB components to produce bioethanol and other high value chemicals.
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.
Production of succinic acid via separate enzymatic hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) are alternatives and are environmentally friendly processes. These processes have attained considerable positions in the industry with their own share of challenges and problems. The high-value succinic acid is extensively used in chemical, food, pharmaceutical, leather and textile industries and can be efficiently produced via several methods. Previously, succinic acid production via chemical synthesis from petrochemical or refined sugar has been the focus of interest of most reviewers. However, these expensive substrates have been recently replaced by alternative sustainable raw materials such as lignocellulosic biomass, which is cheap and abundantly available. Thus, this review focuses on succinic acid production utilizing lignocellulosic material as a potential substrate for SSF and SHF. SSF is an economical single-step process which can be a substitute for SHF - a two-step process where biomass is hydrolyzed in the first step and fermented in the second step. SSF of lignocellulosic biomass under optimum temperature and pH conditions results in the controlled release of sugar and simultaneous conversion into succinic acid by specific microorganisms, reducing reaction time and costs and increasing productivity. In addition, main process parameters which influence SHF and SSF processes such as batch and fed-batch fermentation conditions using different microbial strains are discussed in detail.
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.
As one of Lianyungang's most famous specialties, Acanthogobius hasta is delicious and nutritious fish, but is extremely susceptible to spoilage during transportation and storage. In this study, Lactobacillus plantarum MMB-07 was isolated from traditional fermented sour fish to reduce losses and improve the utilization and food value of A. hasta. L. plantarum MMB-07 had good ability of acid production and acid resistance. Moreover, it could also inhibit common pathogens in food or aquatic products to ensure the safety of fermented products. MMB-07 was used to ferment A. hasta and obtain fermented Suanyu rich in nutrition value and good flavor. The volatile base nitrogen was 18.44 mg/100 g and the fermented fish meat maintained second-grade freshness. Thiobarbituric acid assay was 0.90 mg/kg and fat in fish meat was oxidized to a low degree. The studies indicated that MMB-07 has a high application prospect in low salt fermented fish.
The yield and properties of cellulose produced from bacterial fermentation of black tea broth (known as Kombucha) were investigated in this study. The tea broth was fermented naturally over a period of up to 8 days in the presence of sucrose. Tea broth with a sucrose concentration of 90 g/l produced highest yield of bacterial cellulose (66.9%). The thickness and yield of bacterial cellulose increased with fermentation time. The bacterial cellulose production increased correspondingly with increased surface area:depth ratio. Changes in pH were related to the symbiotic metabolic activities of yeasts and acetic acid bacteria, and the counts of both of these in the tea broths were relatively higher than those in the cellulose layer. Findings from this study suggest that the yield of cellulose depends on many factors that need to be optimized to achieve maximum yield.
Virgin Coconut Oils (VCO) were prepared from fresh-dry (grated coconut route), chilling and thawing, enzymatic and fermentation method in this study. All of the VCO produced conformed physicochemically to the standards established by the Asian and Pacific Coconut Community (APCC) and Codex Alimentarius Commission. The highest FA (fatty acid) is lauric acid in all of the VCO and ranged from 46.36 – 48.42 %, while the principal TAG (triacylglycerol) is LaLaLa (La: Lauric) with 17.94 – 19.83 % of the total TAG. Tocopherol analysis showed the presence of beta, gamma and delta tocopherols at low levels. In all, the physicochemical, FA and TAG analyses of the VCO extracted from different methods showed some significant differences, while the tocopherol content does not differ significantly among the different types of extraction methods used.
Cocoa pod borer (Conopomorpha cramerella (Snellen)) is the main fatal pest that destroys cocoa plantations in South East Asia, mainly in Indonesia and Malaysia. Infested cocoa beans stick to each other, the pulp become hard and normal fermentation process to produce flavour precursors cannot be done. This research aimed to utilize the infested cocoa beans as a source of phenolic compounds. Extraction of phenolic compounds was carried out for three infestation levels regarded as low, medium and heavy. Parameters of study were bean size, shell content, fat content, total polyphenol and antioxidant activity. Results of the study showed that the increase in cocoa pod borer infestation significantly decreased cocoa bean size, fat content and total polyphenol; but shell content was increased. Antioxidant activity of the extracted polyphenol was not significantly affected by the infestation. The results clearly indicate that cocoa beans infested by cocoa pod borer can potentially to be used as a source of phenolic compounds for natural antioxidant uses.
The effects of lactic acid bacteria (Lactobacillus plantarum and Lactobacillus bulgaricus) inoculation
on the sensory attributes and consumers acceptance of fermented curry paste compared with uncultured sample were assessed. pH, titratable acidity (TA) and color changes, during four-month storage were monitored. Hedonic test was utilized to evaluate consumer perception and acceptability of fermented and ordinary curry pastes. Rapid pH drop was observed in inoculated sample with Lb. plantarum presenting better performance than the Lb. bulgaricus. Titratable acidity increased significantly (p0.05) in most of the attributes of
original recipe and fermented curry paste except for color and sweetness. In summary, this study showed fermented curry paste with Lb. plantarum and Lb. bulgaricus exhibited new sensory attributes encouraging acceptability by consumers.
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
.
Xylitol can be obtained from lignocellulosic materials containing xylose. However, the fraction of lignocellulose converted through dilute acid hydrolysis contains compounds that inhibit the fermenting micro-organisms. These inhibitors can be removed from the hydrolysate by detoxification method, prior to fermentation. This study describes effectiveness of overliming process to reduce the toxicity of hydrolysates generated from pre-treatment of sago trunk for xylitol production. The overliming pH 9 and 10 was studied and the results showed that pH 9 was showed 20% of sugar loss, which is low compared to pH 10. Candida tropicalis strain was used to evaluate the fermentability of overlimed sago trunk hydrolysate at pH 9 and non-overlimed hydrolysate medium. Meanwhile, Xylitol accumulation and productivity in the overlimed medium was found to be higher than the non-treated medium. The maximum production of xylitol was increased up to 74% and converted within 76 h. The results obtained improved the fermentation process when compared with the nontreated medium.
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.
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.
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.
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.
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.