Efficient production of lactic acid from cellulose and xylan in sugarcane bagasse by newly isolated Lactiplantibacillus plantarum and Levilactobacillus brevis through simultaneous saccharification and co-fermentation process.

Sugarcane bagasse is one of the promising lignocellulosic feedstocks for bio-based chemicals production. However, to date, most research focuses mainly on the cellulose conversion process, while hemicellulose remains largely underutilized. The conversion of glucose and xylose derived from lignocellulosic biomass can be a promising strategy to improve utilization efficiencies of resources, energy, and water, and at the same time reduce wastes generated from the process. Here, attempts were made to convert cellulose and xylan in sugarcane bagasse (SB) into lactic acid (LA) through a pre-hydrolysis and simultaneous saccharification and co-fermentation (SScF) process using newly isolated Lactiplantibacillus plantarum TSKKU P-8 and Levilactobacillus brevis CHKKU N-6. The process yielded 91.9 g/L of LA, with a volumetric productivity of 0.85 g/(L·h). This was equivalent to 137.8 ± 3.4 g-LA, a yield on substrate (pretreated SB) of 0.86 g/g, and a productivity of 1.28 g/h, based on a final volume of 1.5 L. On the other hand, pre-hydrolysis and simultaneous saccharification and fermentation (SSF) process using La. plantarum TSKKU P-8 as a monoculture gave 86.7 ± 0.2 g/L of LA and a volumetric productivity of 0.8 g/(L·h), which were equivalent to 104.8 ± 0.3 g-LA, a yield on substrate of 0.65 g/g, and a productivity of 0.97 g/h, based on a final volume of 1.2 L. Mass balance calculated based on mass of raw SB entering the process showed that the SScF process improved the product yield by 32% as compared with SSF process, resulting in 14% improvement in medium-based economic yield.

High Cell Density Cultivation of Paracoccus sp. on Sugarcane Juice for Poly(3-hydroxybutyrate) Production.

High cell density cultivation is a promising approach to reduce capital and operating costs of poly (3-hydroxybutyrate) (PHB) production. To achieve high cell concentration, it is necessary that the cultivation conditions are adjusted and controlled to support the best growth of the PHB producer. In the present study, carbon to nitrogen (C/N) ratio of a sugarcane juice (SJ)-based medium, initial sugar concentration, and dissolved oxygen (DO) set point, were optimized for batch cultivation of Paracoccus sp. KKU01. A maximum biomass concentration of 55.5 g/L was attained using the C/N ratio of 10, initial sugar concentration of 100 g/L, and 20% DO set point. Fed-batch cultivation conducted under these optimum conditions, with two feedings of SJ-based medium, gave the final cell concentration of 87.9 g/L, with a PHB content, concentration, and yield of 36.2%, 32.1 g/L, and 0.13 g/g-sugar, respectively. A medium-based economic analysis showed that the economic yield of PHB on nutrients was 0.14. These results reveal the possibility of using SJ for high cell density cultivation of Paracoccus sp. KKU01 for PHB production. However, further optimization of the process is necessary to make it more efficient and cost-effective.

Improvement of hydrogen production from Chlorella sp. biomass by acid-thermal pretreatment.

BACKGROUND: Owing to the high growth rate, high protein and carbohydrate contents, and an ability to grow autotrophically, microalgal biomass is regarded as a promising feedstock for fermentative hydrogen production. However, the rigid cell wall of microalgae impedes efficient hydrolysis of the biomass, resulting in low availability of assimilable nutrients and, consequently, low hydrogen production. Therefore, pretreatment of the biomass is necessary in order to achieve higher hydrogen yield (HY). In the present study, acid-thermal pretreatment of Chlorella sp. biomass was investigated. Conditions for the pretreatment, as well as those for hydrogen production from the pretreated biomass, were optimized. Acid pretreatment was also conducted for comparison. RESULTS: Under optimum conditions (0.75% (v/v) H2SO4, 160 °C, 30 min, and 40 g-biomass/L), acid-thermal pretreatment yielded 151.8 mg-reducing-sugar/g-biomass. This was around 15 times that obtained from the acid pretreatment under optimum conditions (4% (v/v) H2SO4, 150 min, and 40 g-biomass/L). Fermentation of the acid-thermal pretreated biomass gave 1,079 mL-H2/L, with a HY of 54.0 mL-H2/g-volatile-solids (VS), while only 394 mL/L and 26.3 mL-H2/g-VS were obtained from the acid-pretreated biomass. CONCLUSIONS: Acid-thermal pretreatment was effective in solubilizing the biomass of Chlorella sp. Heat exerted synergistic effect with acid to release nutrients from the biomass. Satisfactory HY obtained with the acid-thermal pretreated biomass demonstrates that this pretreatment method was effective, and that it should be implemented to achieve high HY.

Characterization of vacuolar amino acid transporter from Fusarium oxysporum in Saccharomyces cerevisiae.

Fusarium oxysporum causes wilt disease in many plant families, and many genes are involved in its development or growth in host plants. A recent study revealed that vacuolar amino acid transporters play an important role in spore formation in Schizosaccharomyces pombe and Saccharomyces cerevisiae. To investigate the role of vacuolar amino acid transporters of this phytopathogenic fungus, the FOXG_11334 (FoAVT3) gene from F. oxysporum was isolated and its function was characterized. Transcription of FoAVT3 was upregulated after rapamycin treatment. A green fluorescent protein fusion of FoAvt3p was localized to vacuolar membranes in both S. cerevisiae and F. oxysporum. Analysis of the amino acid content of the vacuolar fraction and amino acid transport activities using vacuolar membrane vesicles from S. cerevisiae cells heterologously expressing FoAVT3 revealed that FoAvt3p functions as a vacuolar amino acid transporter, exporting neutral amino acids. We conclude that the FoAVT3 gene encodes a vacuolar neutral amino acid transporter.

Functional Expression and Characterization of Schizosaccharomyces pombe Avt3p as a Vacuolar Amino Acid Exporter in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, Avt3p and Avt4p mediate the extrusion of several amino acids from the vacuolar lumen into the cytosol. SpAvt3p of Schizosaccharomyces pombe, a homologue of these vacuolar amino acid transporters, has been indicated to be involved in spore formation. In this study, we confirmed that GFP-SpAvt3p localized to the vacuolar membrane in S. pombe. The amounts of various amino acids increased significantly in the vacuolar pool of avt3Δ cells, but decreased in that of avt3+-overexpressing avt3Δ cells. These results suggest that SpAvt3p participates in the vacuolar compartmentalization of amino acids in S. pombe. To examine the export activity of SpAvt3p, we expressed the avt3+ gene in S. cerevisiae cells. We found that the heterologously overproduced GFP-SpAvt3p localized to the vacuolar membrane in S. cerevisiae. Using the vacuolar membrane vesicles isolated from avt3+-overexpressing S. cerevisiae cells, we detected the export activities of alanine and tyrosine in an ATP-dependent manner. These activities were inhibited by the addition of a V-ATPase inhibitor, concanamycin A, thereby suggesting that the activity of SpAvt3p is dependent on a proton electrochemical gradient generated by the action of V-ATPase. In addition, the amounts of various amino acids in the vacuolar pools of S. cerevisiae cells were decreased by the overproduction of SpAvt3p, which indicated that SpAvt3p was functional in S. cerevisiae cells. Thus, SpAvt3p is a vacuolar transporter that is involved in the export of amino acids from S. pombe vacuoles.