Satoshi Akao

Kinetic model of thermophilic L-lactate fermentation by Bacillus coagulans combined with real-time PCR quantification

A simple L-lactate fermentation of organic wastes at pH 5.5 and 55 degrees C under nonsterile conditions using Bacillus coagulans can be suitable for L-lactate fermentation of garbage. A mathematical model that simulated the lactate fermentation characteristics of B. coagulans was developed by focusing on the inhibitory effects of substrate, lactate (product) and NaCl, and bacterial growth. Basic fermentation experiments were performed using simple substrates to derive fundamental parameters of growth rate and inhibition effects. The model was then applied to fermentations using simple substrates and artificial kitchen garbage in order to verify its applicability. Microbial concentration, a key state variable of the model was measured using both real-time polymerase chain reaction (PCR) and traditional methods. The results of these methods were compared for experimental cases in which only soluble substrates were used. B. coagulans concentrations were suitably measured using real-time PCR, even when traditional measurement methods for microbial concentrations cannot be used. The results indicate that the developed model and biomass measurement can be used to evaluate lactate fermentations using both simple and complex substrates. These proposed methods would be useful for developing a new bacterial function-based mathematical model for more complex acid fermentations.

Nutrient recovery from biomass cultivated as catch crop for removing accumulated fertilizer in farm soil

As a result of long-term continuous use of fertilizers in farm land, a large amount of nutrients accumulate in the soil, increasing the risk of eutrophication or nitrate pollution of groundwater. For rehabilitating the farm soil and recovering nutrients such as nitrogen, phosphorus and potassium, a new system has been developed by our research group. This paper discusses the methodology of extracting nutrients from biomass in order to recover phosphorus and other nutrients in crystal form. Around 80% or higher extraction rates were achieved for phosphorus and potassium by soaking the powdered tissue in distilled water or 1% NaOH solution for 24 h. The extracted phosphorus and potassium act as a potential resource for recycled fertilizer or other industrial materials.

Comparison of simultaneous and separate processes: saccharification and thermophilic L-lactate fermentation of catch crop and aquatic plant biomass.

Catch crop candidates (corn, guinea grass) for recovering nutrients from farm soil and aquatic plants (water caltrop, water hyacinth) were utilized to produce l-lactic acid. The efficiencies of pre-treatment methods for enzymatic saccharification and l-lactate production of two fermentation processes, thermophilic simultaneous saccharification and fermentation (SSF), as well as separate saccharification and fermentation, were compared. Conditions were set at 55°C and pH 5.5 for non-sterile fermentation. Alkaline/peroxide pre-treatment proved the most effective for saccharification in pre-treated corn, guinea grass, water caltrop and water hyacinth with glucose yields of 0.23, 0.20, 0.11 and 0.14 g/g-dry native biomass (24-hour incubation period), respectively. Examination of the two types of thermophilic l-lactate fermentation employed following alkaline/peroxide pre-treatment and saccharification demonstrated that the l-lactate yield obtained using SSF (0.15 g/g in the case of corn) was lower than that obtained using separate saccharification and fermentation (0.28 g/g in the case of corn). The lower yield obtained from SSF is likely to have resulted from the saccharification conditions used in the present study, as the possibility of cellulase deactivation during SSF by thermophilic l-lactate producing bacteria existed. A cellulase that retains high activity levels under non-sterile conditions and a l-lactate producer without cellulose hydrolysis activity would be required in order for SSF to serve as an effective method of l-lactate production.

Seasonal changes in the performance of a catch crop for mitigating diffuse agricultural pollution

An in situ technology for mitigating diffuse agricultural pollution using catch crops was developed for simultaneously preventing nitrate groundwater pollution, reducing nitrous oxide (N2O) gas emissions, and removing salts from the topsoil. Seasonal changes in the performance of a catch crop were investigated using lysimeters in a full-scale greenhouse experiment with 50 d cultivation of dent corn. Catch crop cultivation significantly reduced the leached mineral nitrogen by 89-91% in summer, 87-89% in spring, and 61-82% in winter, and it also significantly reduced the N2O emission by 68-84% in summer. The amounts of nitrogen uptake by the catch crop were remarkably higher than those of leached nitrogen and N2O emission in each season. Catch crop cultivation is a promising technology for mitigating diffuse agricultural pollution.

Effects of soil type and nitrate concentration on denitrification products (N2O and N2) under flooded conditions in laboratory microcosms

Abstract Denitrification products nitrous oxide ((N2O) and nitrogen (N2)) were measured in three flooded soils (paddy soil from Vietnam, PV; mangrove soil from Vietnam, MV; paddy soil from Japan, PJ) with different nitrate (NO3–) concentrations. Closed incubation experiments were conducted in 100-mL bottles for 7 d at 25°C. Each bottle contained 2 g of air-dried soil and 25 mL solution with NO3– (concentration 0, 5 or 10 mg N L−1) with or without acetylene (C2H2). The N2O + N2 emissions were estimated by the C2H2 inhibition method. Results showed that N2O + N2 emissions for 7 d were positively correlated with those of NO3– removal from solution with C2H2 (R2 = 0.9872), indicating that most removed NO3– was transformed to N2O and N2 by denitrification. In PJ soil, N2O and N2 emissions were increased significantly (P < 0.05) by the addition of greater NO3– concentrations. However, N2O and N2 emissions from PV and MV soils were increased by the addition of 0 to 5 mg N L−1, but not by 5 to 10 mg N L−1. At 10 mg N L−1, N2 emissions for 7 d were greater in PJ soil (pH 7.0) than in PV (pH 5.8) or MV (pH 4.3) soils, while N2O emissions were higher in PV and MV soils than in PJ soil. In MV soil, N2O was the main product throughout the experiment. In conclusion, NO3– concentration and soil pH affected N2O and N2 emissions from three flooded soils.

Nitrous oxide emissions during biological soil disinfestation with different organic matter and plastic mulch films in laboratory-scale tests.

Nitrous oxide (N2O), which is a greenhouse gas, may be more emitted as an intermediate product of denitrification during biological soil disinfestation. The biological soil disinfestation is a method to suppress soil-borne pathogens under reductive soil conditions produced by the application of organic matter and water irrigation with plastic film. The objective of the study was to determine the effects of different organic matter and mulch films on N2O emissions during biological soil disinfestation. Grey lowland soil amended with cattle compost plus rice bran (0.2%), rice husk (0.2%) or dent corn (0.1%, 0.2% and 0.4%) was incubated at 100% water-holding capacity with or without plastic films made of polyvinyl chloride (PVC) and triple-layer polyolefin (3PO) for 72 h at 50°C. Permeation of the two films was also measured at 25°C and 50°C. Results showed that incorporation of organic matter increased N2O emissions compared with no organic matter addition at 50°C. Incorporation of rice bran and dent corn with easily decomposable C and low C:N ratios increased N2O emissions for the first 12 h, but thereafter, available C supply from these amendments suppressed N2O emissions. Permeability of mulch films increased at a higher temperature and was larger for PVC than for 3PO. Our study indicated that rice husk should not be used for soil disinfestation and that application rates of organic matter must be determined based on their decomposability. Moreover, mulch film covering would not suppress N2O emission in biological soil disinfestation because of high temperature.