Promoting Magnusiomyces spicifer AW2 Cell-Bound Lipase Production by Co-culturing with Staphylococcus hominis AUP19 and Its Application in Solvent-Free Biodiesel Synthesis.

Yeast-bacterium interaction has recently been investigated to benefit the production of cell-bound lipases (CBLs). Staphylococcus hominis AUP19 supported the growth of Magnusiomyces spicifer AW2 in a palm oil mill effluent (POME) medium to produce CBLs through a bioremediation approach, including oil and grease (O&G) and chemical oxygen demand (COD) removals. This research used the yeast-bacterium co-culture to optimize CBLs and cell biomass (CBM) productions through bioremediation using the statistical Plackett-Burman design and response surface methodology-central composite design. The CBLs were finally applied in biodiesel synthesis. The CBM of 13.8 g/L with CBLs activity at 3391 U/L was achieved after incubation at room temperature (RT, 30 ± 2 °C) for 140 h in 50% POME medium, pH 7.0, containing 1.23% (w/v) ammonium sulfate. Bacterium promoted yeast growth to achieve bioremediation with 87.9% O&G removal and 84.5% COD removal. Time course study showed that the CBLs activity was highest at 24 h cultivation (4103 U/L) and retained 80% and 60% of activities at 4 °C and RT after 5 weeks of storage. The CBLs application successfully yielded 77.3% biodiesel from oleic acid (esterification) and 86.4% biodiesel from palm oil (transesterification) within 72 h in solvent-free systems. This study highlights that yeast-bacterium co-culture and POME should receive more attention for potential low-cost CBLs production through bioremediation, i.e., O&G and COD removals, while the CBLs as biocatalysts are promising for significant contribution to an effective strategy for economic green biodiesel production.

Dual-bioaugmentation strategy to enhance the formation of algal-bacteria symbiosis biofloc in aquaculture wastewater supplemented with agricultural wastes as an alternative nutrient sources and biomass support materials.

This study performs an integrated evaluation of the formation and distribution of algal-bacterial bioflocs in aquaculture wastewater supplemented with agricultural waste, together with an assessment of their behavior in the microbial community and of the water quality of the system in which a new bioaugmentation strategy was applied. Results indicated that the dual bioaugmentation strategy via the consortium addition of bacteria and microalgae had the highest formation performance, providing the most compact biofloc structure (0.59 g/L), excellent settleability (71.91%), and a large particle diameter (4.25 mm). The fed-batch supplementation of molasses and rice bran, in terms of changes in the values of COD, NH4+, NO3-, and PO43-, stimulated the formation of biofloc through algal-bacterial bioflocs and microbe-rice bran complexes within a well-established microbial community. These findings provide new insight into the influence of bioaugmentation on the formation of an innovative algal-bacterial biofloc.

Lipid Profile, Antioxidant and Antihypertensive Activity, and Computational Molecular Docking of Diatom Fatty Acids as ACE Inhibitors.

Diatoms, as single cell eukaryotic microalgae, are rich sources of lipids, which have either beneficial or detrimental effects on the prevention and treatment of many diseases. Gas chromatography-mass spectrometry (GC-MS) identified diatom lipids with high levels of essential fatty acids (EFAs), especially polyunsaturated FAs (PUFAs) containing both omega-3 and omega-6. Nutritional values of FAs indicated possible applications in the pharmaceutical, nutraceutical, and functional food industries. Diatom FAs showed antioxidative potential on harmful radicals by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) scavenging, with high inhibition of the angiotensin-converting enzyme (ACE) that causes cardiovascular disease (CVD) and hypertension. A computational molecular docking simulation confirmed the inhibition mechanisms of FAs on ACE, with comparable levels of binding free energy to chemically synthesized ACE drugs. Findings suggested that diatom lipids showed potential for use as alternative ACE inhibitors or food supplement for CVD prevention.

Application of palm oil mill waste to enhance biogas upgrading and hornwort cultivation.

The potential of oil palm ash (OPA) to enhance H2S and CO2 removal from biogas by scrubbing with maturation pond effluent (MPE), and further the treatment of biogas scrubber effluent (BSE) by Ceratophyllum demersum L. (hornwort) cultivation were investigated in this study. The results show that OPA + MPE solution with pH 9.3 and alkalinity 7525 mg CaCO3/L was obtained with 0.7 kg/L OPA loading. A pilot scale scrubber was used to study the effects of absorbent flow rates of 60-210 L/h on upgrading to 300 L/h field biogas stream. At 210 L/h, the CO2 removal efficiencies were 33% and 53% for MPE and OPA + MPE, respectively. To approach 100% H2S removal efficiency, the minimum flow rates were 120 L/h for MPE and 90 L/h for OPA + MPE. 50-150 g wet weight of hornwort in 30 L diluted POME were loaded to investigate appropriate initial hornwort loading level for hornwort cultivation. The highest specific growth rate of 0.045 day-1 with biomass production of 3.8 g/day were obtained with a 50 g initial loading. Among the wastewaters (MPE, OPA + MPE, and BSE) treatment using hornwort cultivation, the highest 0.035 day-1 specific growth rate and 2.6 g/day biomass production of hornwort were obtained in diluted BSE cultivation, and in 3 weeks of cultivation. COD, nitrate, phosphate, and alkalinity decreased by 76%, 76%, 55%, and 5%, respectively. The Eco-Efficiency concept for palm oil mill waste utilization proposed in this study has a high potential for enhanced biogas upgrading by using OPA + MPE, and hornwort is a good candidate for BSE post-treatment integrated with biomass production.

Potential use of industrial by-products as promising feedstock for microbial lipid and lipase production and direct transesterification of wet yeast into biodiesel by lipase and acid catalysts.

This work attempted the conversion of crude glycerol to lipid and lipase by Yarrowia lipolytica and the direct transesterification of wet yeast by its lipase into biodiesel via response surface methodology to enhance the cost-effectiveness of biodiesel production from the lipids. The yeast grew better and accumulated a high amount of lipids on the waste combined with fish waste hydrolysate, but only exhibited high lipase activity on the waste supplemented with surfactants (i.e., gum Arabic, Tween 20, Tween 80). However, the combination of both wastes and Tween 80 further improved growth, lipid productivity, and lipase activity. More importantly, lipase-direct transesterification under optimal conditions (wet cell concentration of 17.97 mg-DCW, methanol loading of 8.21 µL, and hexane loading of 10.26 µL) followed by acid-catalyst transesterification (0.4 M H2SO4), offered high FAME yields (>90%), showing the efficiency of the process when applied for the industrialization of biodiesel production from microbial lipids.

Utilization of palm oil mill effluent as a novel substrate for the production of antifungal compounds by Streptomyces philanthi RM-1-138 and evaluation of its efficacy in suppression of three strains of oil palm pathogen.

AIMS: This study aimed to use palm oil mill effluent (POME) as a renewable resource for the production of antifungal compounds by Streptomyces philanthi RM-1-138 against Ganoderma boninense, Ceratocystis paradoxa and Curvularia oryzae. METHODS AND RESULTS: The efficacy of antifungal compounds RM-1-138 against the three strains of fungal oil palm pathogen was evaluated both in vitro and on oil palm leaf segments. In vitro studies using confrontation tests on glucose yeast-malt extract (GYM) agar plates indicated that the strain RM-1-138 inhibited the growth of all three fungal pathogenic strains. The antifungal compounds produced in the GYM medium exhibited significantly higher inhibition (79%-100%) against the three fungal pathogens than using the diluted POME (50%) medium (80%-83% inhibition). The optimum condition for the production of antifungal compounds from the strain RM-1-138 was as following: POME of 47,966 mg L-1 chemical oxygen demand (COD), the initial pH at 7.0 and supplemented with yeast extract (0.4%). Meanwhile, severe morphological and internal abnormalities in C. oryzae hyphae were observed under a scanning electron microscope and transmission electron microscope. In vivo experiment on oil palm leaf segments indicated that the efficacy of the antifungal compounds RM-1-138 (DSI = 1.3) were not significantly difference in the suppression of Curvularia leaf spot compared with the two commercial chemical fungicides of mancozeb® (DSI = 1.0) and tetraconazole® (DSI = 1.3). CONCLUSIONS: Antifungal compounds produced by S. philanthi RM-1-138 grown in POME have the potential to inhibit fungal pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The POME (about 47 mg L-1 COD) with the initial pH of 7.0 and supplementation of 0.4% nitrogen could be used as a culture medium for the growth and production of antifungal compounds of S. philanthi RL-1-138. In addition, the antifungal compound RM-1-138 could suppress the three strains of oil palm fungal pathogen tested on oil palm leaf segment.

Catalytic pyrolysis of petroleum-based and biodegradable plastic waste to obtain high-value chemicals.

The petroleum-based plastics, high-density polyethylene (HDPE), low-density polyethylene (LDPE), and polypropylene (PP), and the biodegradable plastic, polylactide (PLA) were processed by thermal and catalytic pyrolysis to investigate their suitability as feedstock for chemical recycling. The influence of pyrolysis temperature (400-600 °C) and catalyst (zeolite, spent FCC, and MgO catalyst) on the pyrolysis liquid composition and yield was studied. The studied petroleum-based plastics had similar decomposition temperature ranges but produced their highest pyrolysis yields at different temperatures. Pyrolysis liquids from thermal degradation of HDPE and LDPE consisted high yield of waxes but those of PP and PLA consisted of both waxes and liquid oil. Catalysts affected not only the pyrolysis yield, but also the proportions of liquid oil and wax in pyrolysis liquids. Alkenes, alkanes, and aromatics were the main compounds in the pyrolysis liquids. Spent FCC catalyst reduced the production of waxes and increased the production of gasoline-range hydrocarbons and aromatics. MgO catalyst led to high coke formation from polyolefins and PLA. Lactic acid, lactide and propanoic acid were examples of valuable chemicals recovered from the pyrolysis of PLA. Lactide was the main product (up to 79%) of catalytic pyrolysis with zeolite at 400 °C. Spent FCC catalyst produced mostly propanoic acid at 400 °C but at 600 °C, L-lactic acid became the most abundant compound.

The Occurrence of Triple Catalytic Characteristics of Yeast Lipases and Their Application Prospects in Biodiesel Production from Non-Edible Jatropha curcas Oil in a Solvent-Free System.

Extracellular and cell-bound lipase-producing yeasts were isolated from the palm oil mill wastes and investigated for their potential uses as biocatalysts in biodiesel production. Twenty-six yeast strains were qualitatively screened as lipase producers. From those yeast strains, only six were selected and screened further for quantitative lipase production.The phylogenetic affiliations of the yeast strains were confirmed by investigating the D1/D2 domains of 26S rDNA and ITS1-5.8S-ITS2 molecular regions of the six yeast strains selected as potent lipase producers. The three yeast strains A4C, 18B, and 10F showed a close association with Magnusiomyces capitatus. Two yeast strains (17B and AgB) had a close relationship with Saprochaete clavata, whereas the strain AW2 was identified as Magnusiomyces spicifer. Three main catalytic activities of the yeast lipases were evaluated and Magnusiomyces capitatus A4C, among the selected lipase-producing yeasts, had the highest extracellular lipolytic enzyme activity (969 U/L) with the cell-bound lipolytic enzyme activity of 11.3 U/gdm. The maximum cell-bound lipolytic activity (12.4 U/gdm) was observed in the cell-bound lipase fraction produced by Magnusiomyces spicifer AW2 with an extracellular lipolytic enzyme activity of 886 U/L. Based on the specific hydrolytic enzymatic activities, the cell-bound lipases (CBLs) from the three yeast strains M. capitatus A4C, M. spicifer AW2, and Saprochaete clavata 17B were further investigated for biodiesel production. Among them, the CBL from M. spicifer AW2 synthesized the most FAME (fatty acid methyl esters) at 81.2% within 12 h indicating that it has potential for application in enzymatic biodiesel production.

Valorization of palm biomass wastes for biodiesel feedstock and clean solid biofuel through non-sterile repeated solid-state fermentation.

Palm biomass wastes are currently considered as promising solid biofuels. However, their high potassium content leads to formation of slag in combustion chambers and causes frequent power-plant shutdowns for maintenance. Therefore, this study aimed to develop a low-cost practical biological pretreatment for these wastes. Oleaginous fungi Aspergillus tubingensis TSIP9, which originates from palm wastes, was used to pretreat biomass wastes and simultaneously produce oils through non-sterile solid state fermentation (SoSF). The operating conditions were optimized through response surface methodology. The fungi could grow and produce oils with good biodiesel fuel properties. After SoSF, potassium content in biomass wastes was reduced by 90% and cellulose content increased to >57%, making it suitable as clean solid biofuel. Repeated-SoSF with 90% substrate replacement was highly effective in continuously pretreating biomass wastes and producing fungal oils. This study demonstrates the cost-effective and environmentally friendly process for production of clean renewable energy through zero-waste strategy.

Immobilized oleaginous microalgae as effective two-phase purify unit for biogas and anaerobic digester effluent coupling with lipid production.

Oleaginous microalga Scenedesmus sp. was immobilized in alginate-gel beads and applied as two-phase purify unit for biogas and anaerobic digester effluent from palm oil mill. Optimal microalgal cell concentration and bead volume ratio were 106 cells mL-1 and 25% v/v, respectively. The use of 20% effluent and light intensity at 128 µmol·proton·m-2 s-1 most promoted CO2 removal by immobilized microalgae and achieved the maximum CO2 removal rate of 4.63 kg-CO2 day-1 m-3. This process upgraded methane content in biogas (>95%) and completely remove nitrogen and phosphorus in the effluent. After process operation, 2.98 g L-1 microalgal biomass with 35.92% lipid content were recovered by simple sieving method. Microalgal lipids are composed of C16-C18 (>98%) with prospect high cetane number and short ignition delay time. This study has shown the promising biorefinery concept which is effective not only in CO2 fixation, biogas upgrading and pollutant removal but also cost-effective production of microalgae-based biofuel.

Immobilized oleaginous microalgae for production of lipid and phytoremediation of secondary effluent from palm oil mill in fluidized bed photobioreactor.

Oleaginous microalga Nannochloropsis sp. was immobilized in alginate gel beads and cultivated under optimal conditions that their growth and lipid production were comparable to those of free cells. The immobilized cells were used in phytoremediation of secondary effluent from palm oil mill and easily recovered by simple sieving method. The immobilized cells contributed to removal of nitrogen and phosphorus >90% and CO2 mitigation >99%. They also gave the biomass and lipid production of 1.300±0.050g/L and 0.356±0.097g/L, respectively. The repeated-batch cultivation improved the biomass and lipid production by 2.66 folds and 1.41 folds, respectively. The scale up in 3L-fluidized bed photobioreactor gave the maximum biomass of 3.280±0.049g/L and lipid production of 0.362±0.010g/L. Fatty acid compositions of Nannochloropsis sp. lipids showed their suitability as biodiesel feedstocks. This system not only contributes as tertiary treatment of industrial effluent and CO2 mitigation but also low-cost production of renewable energy.

Felled oil palm trunk as a renewable source for biobutanol production by Clostridium spp.

This study aimed to convert felled oil palm trunk to biobutanol by Clostridium spp. For efficient utilization of oil palm trunk, it was separated into sap and trunk fiber. The sap was used directly while the trunk fiber was hydrolyzed to fermentable sugars before use. Among five clostridia strains screened, Clostridium acetobutylicum DSM 1731 was the most suitable strain for butanol production from the sap without any supplementation of nutrients. It produced the highest amount of butanol (14.4 g/L) from the sap (sugar concentration of 50 g/L) with butanol yield of 0.35 g/g. When hydrolysate from the trunk fiber was used as an alternative carbon source (sugar concentration of 30 g/L), of the strains tested Clostridium beijerinckii TISTR 1461 produced the highest amount of butanol (10.0 g/L) with butanol yield of 0.41 g/g. The results presented herein suggest that oil palm trunk is a promising renewable substrate for biobutanol production.

Industrial wastes as a promising renewable source for production of microbial lipid and direct transesterification of the lipid into biodiesel.

Two strategies of converting industrial wastes to microbial lipid and direct transesterification of obtained lipid into biodiesel were attempted. Several oleaginous yeasts were cultivated on industrial wastes. The yeasts grew well on the wastes with low C/N ratio (i.e. serum latex) but accumulated high lipid content only when the wastes had a high C/N ratio (i.e. palm oil mill effluent and crude glycerol). The yeast lipids have similar fatty acid composition to that of plant oil indicating their potential use as biodiesel feedstocks. The combination of these wastes and two-phase cultivation for cell growth and lipid accumulation improved lipid productivity of the selected yeast. The direct transesterification process that eliminates cell drying and lipid extraction steps, gave comparable yield of biodiesel (fatty acid methyl ester >70% within 1h) to that of conventional method. These two successful strategies may contribute greatly to industrializing oil production from microbes and industrial wastes.

Industrial waste utilization for low-cost production of raw material oil through microbial fermentation.

In view of ever-growing demand of biodiesel, there is an urgent need to look for inexpensive and promising renewable raw material oils for its production. In this context, the aim of this study was to evaluate the potential use of industrial wastes for low-cost production of oils through microbial fermentation. Among the strains tested, Yarrowia lipolytica grew best and produced highest lipid when grown on decanter effluent from palm oil mill. When crude glycerol by-product from a biodiesel plant was added into the effluent as a co-substrate, Y. lipolytica produced a higher biomass of 3.21 g/L and a higher amount of lipid of 2.21 g/L which was 68 % of the dry weight. The scale up and process improvement in a 5-L bioreactor increased the biomass and lipid up to 5.53 and 2.81 g/L, respectively. A semi-continuous mode of operation was an effective mode for biomass enhancement while a fed-batch mode was effective for lipid enhancement. These yeast lipids have potential to be used as biodiesel feedstocks because of their similar fatty acid composition to that of plant oil.

Mixed culture of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris for lipid production from industrial wastes and its use as biodiesel feedstock.

A mixed culture of oleaginous yeast Rhodotorula glutinis and microalga Chlorella vulgaris was performed to enhance lipid production from industrial wastes. These included effluent from seafood processing plant and molasses from sugar cane plant. In the mixed culture, the yeast grew faster and the lipid production was higher than that in the pure cultures. This could be because microalga acted as an oxygen generator for yeast, while yeast provided CO(2) to microalga and both carried out the production of lipids. The optimal conditions for lipid production by the mixed culture were as follows: ratio of yeast to microalga at 1:1; initial pH at 5.0; molasses concentration at 1%; shaking speed at 200 rpm; and light intensity at 5.0 klux under 16:8 hours light and dark cycles. Under these conditions, the highest biomass of 4.63±0.15 g/L and lipid production of 2.88±0.16 g/L were obtained after five days of cultivation. In addition, the plant oil-like fatty acid composition of yeast and microalgal lipids suggested their high potential for use as biodiesel feedstock.

Effect of nitrogen, salt, and iron content in the growth medium and light intensity on lipid production by microalgae isolated from freshwater sources in Thailand.

Four green microalgae (TRG, KB, SK, and PSU) identified as Botryococcus spp. by morphological criteria were isolated from lakes and freshwater ponds in southern Thailand. In nitrogen-rich medium the strains achieved a lipid content of 25.8%, 17.8%, 15.8% and 5.7%, respectively. A combination of nitrogen deficiency, moderately high light intensity (82.5 µE m(-2) s(-1)) and high level of iron (0.74 mM) improved lipid accumulation in TRG, KB, SK, and PSU strains up to 35.9%, 30.2%, 28.4% and 14.7%, respectively. The lipid contents and plant oil-like fatty acid composition of the microalgae suggested their potential as biodiesel feedstock.