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This study was aimed at enhancing galactose consumption from the red seaweed Kappaphycus alvarezii. The optimal pretreatment condition of thermal acid hydrolysis was treated with 350 mM HNO3 for 60 min at 121 °C. The enzymatic saccharification with a 1:1 mixture of Celluclast 1.5 L and Viscozyme L showed the maximum yield of glucose; 42-g/L monosaccharide concentration was obtained with the highest yield of pretreatment and enzymatic saccharification (EPS) and the lowest inhibitory compound concentration. The deletion of the GAL80, MIG1, CYC8, or TUP1 gene was performed to improve the galactose consumption rate. The strains with the deletion of the MIG1 gene (mig1Δ) showed higher galactose consumption rate and ethanol yield than other strains. High transcription levels of regulatory genes revealed that the mig1Δ relieved glucose repression. These results show that the mig1Δ enhances galactose consumption rate from K. alvarezii.

A total 42.68 g/L monosaccharide with 0.10 g/L HMF was obtained from 10% (w/v) Kappaphycus alvarezii with thermal acid hydrolysis using 350 mM HNO3 at 121 °C for 60 min and enzymatic saccharification with a 1:1 mixture of Viscozyme L and Celluclast 1.5 L for 72 h. To enhance the galactose utilization rate, fermentation was performed with overexpression of GAL1 (galactokinase), GAL7 (galactose-1-phosphate uridyltransferase), GAL10 (UDP-glucose-4-epimerase), and PGM2 (phosphoglucomutase 2) in Saccharomyces cerevisiae CEN.PK2 using CCW12 as a strong promoter. Among the strains, the overexpression of PGM2 showed twofold high galactose utilization rate (URgal) and produced ethanol 1.4-fold more than that of the control. Transcriptional analysis revealed the increase of PGM2 transcription level leading to enhance glucose-6-phosphate and fructose-6-phosphate and plays a key role in ensuring a higher glycolytic flux in the PGM2 strain. This finding shows particular importance in biofuel production from seaweed because galactose is one of the major monosaccharides in seaweeds such as K. alvarezii.

Glucose and galactose are monosaccharides obtained from Gloiopeltis furcata (Red algae). A total monosaccharide yield of 62.3 g/L was obtained from G. furcata using thermal acid hydrolysis and enzymatic saccharification. Activated carbon was used to eliminate hydroxymethylfurfural (HMF) from the hydrolysate. Previously obtained monosaccharides are used for ethanol production by Saccharomyces cerevisiae. S. cerevisiae consumes glucose first, then galactose. The methods for reducing fermentation time and increasing the ethanol yield coefficient using the simultaneous consumption of glucose and galactose have been evaluated. Gal3p and Gal80p of S. cerevisiae act as signal transducers that govern the galactose inducer Gal4p mediated transcriptional activation of the Gal gene family. Gal80p binds to Gal4p for transcription deactivation. Therefore, Gal80p was deleted for Gal4p expression without interruption.

Kariba weed (Salvinia molesta) was used as biomass feedstock for ethanol production by separate hydrolysis and fermentation (SHF). Monosaccharides from Kariba weed hydrolysate were produced using thermal acid hydrolysis, sonication, and enzymatic saccharification. The optimal conditions for thermal acid hydrolysis of 12% (w/v) Kariba weed slurry were evaluated as 200 mM HNO3 at 121 degrees C for 60 min yielding 10.2 g/L monosaccharides. Sonication for 45 min before enzymatic saccharification yielded more monosaccharides to 18.7 g/L. Enzymatic saccharification with 16 U/mL Cellic CTec2 produced 35.4 g/L monosaccharides. Fermentation was performed using Saccharomyces cerevisiae, Kluyveromyces marxianus, or Pichia stipitis with sonicated Kariba weed hydrolysate. The control fermentations were carried out using Kariba weed hydrolysate without sonication. The improvement of ethanol production from sonicated Kariba weed hydrolysate using P. stipitis produced 15.9 g/L ethanol with ethanol yield coefficient Y-EtOH = 0.45, K. marxianus produced 14.7 g/L ethanol with Y-EtOH = 0.41. S. cerevisiae produced the lowest yield of 13.2 g/L ethanol with Y-EtOH = 0.37 as it utilized only glucose not xylose. Sonication of Kariba weed was essential in the ethanol production to enhance the productivity of monosaccharides. P. stipitis was determined as the best yeast species using hydrolysates with the mixture of glucose and xylose to produce ethanol.