Valorization of spent coffee grounds through integrated bioprocess of fermentable sugars, volatile fatty acids, yeast-based single-cell protein and biofuels production.

Recovering nutrients from waste for biological processes aligns with sustainability principles. This study aimed to convert spent coffee grounds (SCG) into valuable products, including fermentable sugars, volatile fatty acids (VFAs), yeast-based single-cell protein and biofuels. Alkaline pretreatment was conducted before enzymatic hydrolysis, in which the pretreated SCG was hydrolyzed with varying enzyme loadings (20-60 filter paper units (FPU)/g-solid) and solid loadings (3-15 % w/v). The hydrolyzed slurry was utilized for VFAs and hydrogen production, yielding high values of 0.66 g/g-volatile solids (VS) and 109 mL/g-VS, respectively, using an enzyme loading of 50 FPU/g-solid and a solid loading of 3 % (w/v). The derived VFAs were used to cultivate a newly isolated yeast, Candida maltosa KKU-ARY2, resulting in an accumulated protein content of 43.7 % and a biomass concentration of 4.6 g/L. This study highlights the conversion of SCG into essential components, emphasizing the benefits of waste utilization through cascade bioprocesses.

Microalga Coelastrella sp. Cultivation on Unhydrolyzed Molasses-Based Medium towards the Optimization of Conditions for Growth and Biomass Production under Mixotrophic Cultivation.

Improving biomass production with the utilization of low-cost substrate is a crucial approach to overcome the hindrance of high cost in developing large-scale microalgae production. The microalga Coelastrella sp. KKU-P1 was mixotrophically cultivated using unhydrolyzed molasses as a carbon source, with the key environmental conditions being varied in order to maximize biomass production. The batch cultivation in flasks achieved the highest biomass production of 3.81 g/L, under an initial pH 5.0, a substrate to inoculum ratio of 100:3, an initial total sugar concentration of 10 g/L, and a sodium nitrate concentration of 1.5 g/L with continuous light illumination at 23.7 W/m2. The photobioreactor cultivation results indicated that CO2 supplementation did not improve biomass production. An ambient concentration of CO2 was sufficient to promote the mixotrophic growth of the microalga as indicated by the highest biomass production of 4.28 g/L with 33.91% protein, 46.71% carbohydrate, and 15.10% lipid. The results of the biochemical composition analysis suggest that the microalgal biomass obtained is promising as a source of essential amino acids and pigments as well as saturated and monounsaturated fatty acids. This research highlights the potential for bioresource production via microalgal mixotrophic cultivation using untreated molasses as a low-cost raw material.

Photoautotrophic and Mixotrophic Cultivation of Polyhydroxyalkanoate-Accumulating Microalgae Consortia Selected under Nitrogen and Phosphate Limitation.

Microalgae consortia were photoautotrophically cultivated in sequencing batch photobioreactors (SBPRs) with an alteration of the normal growth and starvation (nutrient limitation) phases to select consortia capable of polyhydroxyalkanoate (PHA) accumulation. At the steady state of SBPR operation, the obtained microalgae consortia, selected under nitrogen and phosphate limitation, accumulated up to 11.38% and 10.24% of PHA in their biomass, which was identified as poly(3-hydroxybutyrate) (P3HB). Photoautotrophic and mixotrophic batch cultivation of the selected microalgae consortia was conducted to investigate the potential of biomass and PHA production. Sugar source supplementation enhanced the biomass and PHA production, with the highest PHA contents of 10.94 and 6.2%, and cumulative PHA productions of 100 and 130 mg/L, with this being achieved with sugarcane juice under nitrogen and phosphate limitation, respectively. The analysis of other macromolecules during batch cultivation indicated a high content of carbohydrates and lipids under nitrogen limitation, while higher protein contents were detected under phosphate limitation. These results recommended the selected microalgae consortia as potential tools for PHA and bioresource production. The mixed-culture non-sterile cultivation system developed in this study provides valuable information for large-scale microalgal PHA production process development following the biorefinery concept.

Bioaugmentation of Lactobacillus delbrueckii ssp. bulgaricus TISTR 895 to enhance bio-hydrogen production of Rhodobacter sphaeroides KKU-PS5.

BACKGROUND: Bioaugmentation or an addition of the desired microorganisms or specialized microbial strains into the anaerobic digesters can enhance the performance of microbial community in the hydrogen production process. Most of the studies focused on a bioaugmentation of native microorganisms capable of producing hydrogen with the dark-fermentative hydrogen producers while information on bioaugmentation of purple non-sulfur photosynthetic bacteria (PNSB) with lactic acid-producing bacteria (LAB) is still limited. In our study, bioaugmentation of Rhodobacter sphaeroides KKU-PS5 with Lactobacillus delbrueckii ssp. bulgaricus TISTR 895 was conducted as a method to produce hydrogen. Unfortunately, even though well-characterized microorganisms were used in the fermentation system, a cultivation of two different organisms in the same bioreactor was still difficult because of the differences in their metabolic types, optimal conditions, and nutritional requirements. Therefore, evaluation of the physical and chemical factors affecting hydrogen production of PNSB augmented with LAB was conducted using a full factorial design followed by response surface methodology (RSM) with central composite design (CCD). RESULTS: A suitable LAB/PNSB ratio and initial cell concentration were found to be 1/12 (w/w) and 0.15 g/L, respectively. The optimal initial pH, light intensity, and Mo concentration obtained from RSM with CCD were 7.92, 8.37 klux and 0.44 mg/L, respectively. Under these optimal conditions, a cumulative hydrogen production of 3396 ± 66 mL H2/L, a hydrogen production rate (HPR) of 9.1 ± 0.2 mL H2/L h, and a hydrogen yield (HY) of 9.65 ± 0.23 mol H2/mol glucose were obtained. KKU-PS5 augmented with TISTR 895 produced hydrogen from glucose at a relatively high HY, 9.65 ± 0.23 mol H2/mol glucose, i.e., 80 % of the theoretical yield. CONCLUSIONS: The ratio of the strains TISTR 895/KKU-PS5 and their initial cell concentrations affected the rate of lactic acid production and its consumption. A suitable LAB/PNSB ratio and initial cell concentration could balance the lactic acid production rate and its consumption in order to avoid lactic acid accumulation in the fermentation system. Through use of appropriate environmental conditions for bioaugmentation of PNSB with LAB, a hydrogen production could be enhanced.

Poly-ß-hydroxyalkanoates production from cassava starch hydrolysate by Cupriavidus sp. KKU38.

A bacterium capable of accumulating polyhydroxyalkanoates (PHAs), Cupriavidus sp. KKU38 (GenBank Accession no. AM260479), was isolated from cassava starch wastewater of cassava starch manufacturing process. KKU38 can utilize glucose, fructose, maltose and xylose for PHA production. Glucose was the most suitable sugar for PHA production giving the highest PHA content and yield of 73.88% and 0.16 g/g-total sugar consumed, respectively. Lactose and sucrose were not suitable carbon sources for both biomass and PHA productions. PHA production from cassava starch hydrolysate (CSH) by the strain KKU38 was maximized under N-limited conditions (COD:N:P of 100:1:2.43). The moderately high biomass concentrations of 5.97 g/L with PHA content and yield of 61.60% and 0.20 g/g-total sugar consumed, respectively, were obtained under the optimum conditions. The analysis of PHA produced under the optimum conditions by (1)H NMR, (13)C NMR and FT-IR indicated that the produced polymer is homopolymer polyhydroxybutyrate (PHB).

Polyhydroxyalkanoates production from effluent of hydrogen fermentation process by Cupriavidus sp. KKU38.

This study investigated the use of the residual sugar and volatile fatty acids (VFAs) in the effluent of the hydrogen production process to produce polyhydroxyalkanoates (PHAs) by Cupriavidus sp. KKU38 in batch fermentation. VFAs in the effluent were lactic, butyric, acetic and propionic acids with a total VFA concentration of 1725 mg/L. The C/N ratio of effluent was 100:2.5, which is defined as the excess carbon and limited nitrogen condition suitable for PHA production. The experiments were conducted in 250-mL Erlenmeyer flasks with a 100 mL working volume. The inoculum size was 30% (v/v) with the initial number of cells 10(6) cells/mL. Residual sugars and acetic acid in the effluent were the major substrates used to produce PHAs while lactic and butyric acids in the effluent were used for biomass synthesis. The maximum PHA concentration and PHA content obtained were 0.85 g/L and 71.42% (w/w) of dry biomass weight, respectively. After fermentation, carbon oxygen demand (COD) in the effluent was reduced by up to 82.73%.

Bioremediation of carbofuran contaminated soil under saturated condition: soil column study.

Disturbed soil columns, 5.8 cm in diameter and 25 cm in length, were used as a basic model to simulate the movement of carbofuran in rice field soil under saturated conditions. Bioaugmentation using a specific carbofuran degrader, Burkholderia sp. PCL3, in free and immobilized cell forms and biostimulation using rice straw as organic amendment were applied with the aim of enhancing the degradation of carbofuran in soil and to prevent the movement of carbofuran along with the flow through. In the abiotic control and the treatment with only indigenous microorganisms, the mass recovery percentage of carbofuran in the effluent was 52.1 and 22.5%, respectively. The application of bioaugmentation or biostimulation significantly enhanced carbofuran degradation in soil and reduced the movement of carbofuran as indicated by a low mass recovery percentage of carbofuran in the effluent of 14.6-15.5%. A low efficiency of carbofuran removal was obtained from the soil column with bioaugmentation together with biostimulation treatments in which the mass recovery percentage of carbofuran in the effluent was in the range of 22.1-22.6%. Sorption of carbofuran to soil, rice straw and corncob, formation of carbofuran metabolite and colony forming unit (CFU) and pH variation with the time were also investigated during column operation.

lux-Marking and application of carbofuran degrader Burkholderia cepacia PCL3.

A luxAB-mutant of the carbofuran degrading bacterium Burkholderia cepacia PCL3 was successfully constructed with the capability to emit a luminescence signal of 1.6×10(-3)RLUcfu(-1). The mutant has a growth pattern and carbofuran degradation ability similar to PCL3 wild-type. The luminescent emission by PCL3:luxAB1 directly correlated with the metabolic activity of the cells. The optimal pH, temperature and n-decanal concentration for luminescence emission are 7.0, 35°C and 0.01%, respectively. PCL3:luxAB1 was used to assess the toxicity of carbofuran and carbofuran phenol in basal salt medium (BSM) in which the different sensitivity of the cells is dependent on the biomass concentration. With the luciferase system, the degradative fraction of the augmented PCL3:luxAB1 and the difference between the active augmented PCL3:luxAB1 and indigenous microorganisms at the contaminated site could be indicated.