Mechanism of inhibition of alpha-amylase by caffeic acid using in-vitro and in-silico techniques.

Type-two diabetes, characterised by insulin resistance or inadequate insulin production, is prevalent among adults. The α-amylase enzyme contributes to carbohydrate digestion, elevating postprandial glucose levels. Natural compounds like caffeic acid offer a solution. This study investigates α-amylase inhibition via in-vitro and in-silico methods, emphasising the connection between phenolic compounds and antidiabetic efficacy for in-silico analysis. Enzyme kinetics, IC50, and molecular docking examine caffeic acid's inhibitory action on α-amylase, comparing it with gallic acid and acarbose. Caffeic acid outperforms acarbose with an IC50 of 4.505 mg/mL versus 16.81 mg/mL, showcasing strong antidiabetic activity. Caffeic acid's superior 1,1-diphenyl-2-picrylhydrazyl (DPPH) inhibition (90.67%) compared to gallic acid (55.76%) indicates potent antioxidative and antidiabetic properties. Molecular docking reveals hydrogen bonding between caffeic acid and α-amylase. These insights lay the groundwork for phenolic-based diabetic therapies, offering less expensive treatment for diabetes patients.

Impact of light spectra on photo-fermentative biohydrogen production by Rhodobacter sphaeroides KKU-PS1.

Purple non-sulphur bacteria can only capture up to 10 % light spectra and only 1-5 % of light is converted efficiently for biohydrogen production. To enhance light capture and conversion efficiencies, it is necessary to understand the impact of various light spectra on light harvesting pigments. During photo-fermentation, Rhodobacter sphaeroides KKU-PS1 cultivated at 30 °C and 150 rpm under different light spectra has been investigated. Results revealed that red light is more beneficial for biomass accumulation, whereas green light showed the greatest impact on photo-fermentative biohydrogen production. Light conversion efficiency by green light is 2-folds of that under control white light, hence photo-hydrogen productivity is ranked as green > red > orange > violet > blue > yellow. These experimental data demonstrated that green and red lights are essential for photo-hydrogen and biomass productions of R. sphaeroides and a clearer understanding that possibly pave the way for further photosynthetic enhancement research.

Enhancing biohydrogen gas production in anaerobic system via comparative chemical pre-treatment on palm oil mill effluent (POME).

Biological hydrogen production using palm oil mill effluent (POME) as a carbon source through dark fermentation process has been suggested to be a promising bioenergy potential and enacts as alternative renewable energy source. Results have indicated that among various 1.5% (w/v) chemical pre-treatments (sodium hydroxide, NaOH; hydrochloric acid, HCl; sulphuric acid, H2SO4; phosphoric acid, H3PO4 and nitric acid, HNO3) on POME, using H3PO4 would generate maximum biohydrogen production of 0.193 mmol/L/h, which corresponded to a yield of 1.51 mol H2/mol TCconsumed with an initial total soluble carbohydrate concentration of 23.52 g/L. Among H3PO4 concentrations (1%-10%), the soluble carbohydrate content and the biohydrogen produced was highest and increased by 1.70-fold and 2.35-fold respectively at 2.5% (w/v), as compared to untreated POME. The batch fermentation maximum hydrogen production rate and yield of 0.208 mmol/L/h and 1.69 mol H2/mol TCconsumed were achieved at optimum pre-treatment conditions of pH 5.5 and thermophilic temperature (60 °C). This study suggests that chemical pre-treatment approaches manage to produce and improve the carbohydrate utilisation process further. Continuous fermentation in CSTR at the optimum conditions produce heightened 1.5-fold biohydrogen yield for 2.5% H3PO4 at 6 h HRT as compared to batch scale. Bacterial community via next-generation sequencing analysis at optimum HRT (6 h) revealed that Thermoanaerobacterium thermosaccharolyticum registered the highest relative frequency of 20.24%. At the class level, Clostridia, Bacilli, Bacteroidia, Thermoanaerobacteria, and Gammaproteobacteria were identified as the biohydrogen-producing bacteria in the continuous system. Insightful findings from this study suggest the substantial practical utility of dilute chemical pre-treatment in improving biohydrogen production.

Potential Utilisation of Dark-Fermented Palm Oil Mill Effluent in Continuous Production of Biomethane by Self-Granulated Mixed Culture.

Two-stage anaerobic digestion of palm oil mill effluent (POME) is a promising method for converting the waste from the largest agricultural industry in Southeast Asia into a clean and sustainable energy. This study investigates the degradation of acid-rich effluent from the dark fermentation stage for the production of biomethane (BioCH4) in a 30-L continuous stirred-tank reactor (CSTR). The continuous methanogenic process was operated with varied HRTs (10 - 1 day) and OLRs (4.6-40.6 gCOD/L.d-1) under thermophilic conditions. Methanothermobacter sp. was the dominant thermophilic archaea that was responsible for the production rate of 4.3 LCH4/LPOME.d-1 and methane yield of 256.77 LCH4kgCOD at HRT of 2 d, which is the lowest HRT reported in the literature. The process was able to digest 85% and 64% of the initial POME's COD and TSS, respectively. The formation of methane producing granules (MPG) played a pivotal role in sustaining the efficient and productive anaerobic system. We report herein that the anaerobic digestion was not only beneficial in reducing the contaminants in the liquid effluent, but generating BioCH4 gas with a positive net energy gain of 7.6 kJ/gCOD.

Multiple crystallization as a potential strategy for efficient recovery of succinic acid following fermentation with immobilized cells.

This study aimed to enhance the crystallizability of bio-based succinic acid for its efficient recovery while maintaining the end product at the highest purity. Immobilization of Actinobacillus succinogenes was initially evaluated based on three different carriers: volcanic glass, clay pebbles, and silica particles. The adsorption capacity of metabolites with a low concentration (10 g/L) and a high concentration (40 g/L) was investigated. It was demonstrated that clay pebbles adsorbed the least succinic acid (< 11 mg/g clay pebbles). The repeated batch-fermentation trials with immobilized cells highlighted that succinic acid with an average concentration of up to 36.3 g/L with a metabolite-production ratio of 3:1 (succinic acid to by-products) could be attained within 130 h. Subsequently, the purification of succinic acid through crystallization was assessed in terms of pH, temperature, crystallization time, initial succinic acid concentration and multiple recrystallization processes. Increasing the crystallization time from 6 h to 9 h afforded an improvement of 17% in the recovery of succinic acid crystals. Moreover, a fourfold concentration coefficient of the broth yielded the highest purity percentage (99.9%). The crystallization in three consecutive stages at 9 h (with a fourfold concentration coefficient) successfully improved the total recovery percentage of succinic acid from 55.0 to 84.8%.

Cocoa pod husk, a new source of hydrolase enzymes for preparation of cross-linked enzyme aggregate.

Cocoa pod husk (CPH) is a by-product of cocoa production obtained after removing the beans from the fruit. The analysis of CPH has shown that it contains high amounts of protein. This study is aimed to utilize this protein source in hydrolase enzyme production. In this study, seven hydrolase enzymes (amylase, fructosyltransferase, mannanase, glucosidase, glucanase, lipase and protease) were screened from CPH for the first time for feasible industrial production. Among these hydrolases, lipase was chosen for the next steps of experiments as it has a lot of applications in different industries. The extraction of high active lipase from CPH has been done under optimum conditions. The condition that was optimum for the three major factors was achieved using Face centered central composite design (FCCCD) with response surface methodology (RSM) to obtain the highest enzyme activity of crude lipase from CPH. The optimum condition of extraction is used for preparation of cross-linked enzyme aggregate (CLEA). For the production of immobilized biocatalyst, the technique of CLEA is considered as an effective technique for its industrially attractive advantages. Referring to the results of OFAT, CLEA-lipase was prepared in the best condition at the presence of 30 mM ammonium sulphate, 70 mM glutaraldehyde with 0.23 mM Bovine serum albumin as an additive. Immobilization effectively improved the stability of lipase against various organic solvents.

Optimized preparation and characterization of CLEA-lipase from cocoa pod husk.

Cross-linked enzyme aggregate (CLEA) is easily prepared from crude enzyme and has many advantages to the environment and it is considered as an economic method in the context of industrial biocatalysis compared to free enzyme. In this work, a highly active and stable CLEA-lipase from cocoa pod husk (CPH) which is a by-product after removal of cocoa beans, were assayed for their hydrolytic activity and characterized under the optimum condition successfully. Face centered central composite design (FCCCD) under response surface methodology (RSM) was used to get the optimal conditions of the three significant factors (concentration of ammonium sulfate, concentration of glutaraldehyde and concentration of additive) to achieve higher enzyme activity of CLEA. From 20 runs, the highest activity recorded was around 9.407U (83% recovered activity) under the condition of using 20% saturated ammonium sulfate, 60mM glutaraldehyde as cross-linker and 0.17mM bovine serum albumin as feeder. Moreover, the optimal reaction temperature and pH value in enzymatic reaction for both crude enzyme and immobilized were found to be 45°C at pH 8 and 60°C at pH 8.2, respectively. A systematic study of the stability of CLEA and crude enzyme was taken with regards to temperature (25-60°C) and pH (5-10) value and in both factors, CLEA-lipase showed more stability than free lipase. The Km value of CLEA was higher compared to free enzyme (0.55mM vs. 0.08mM). The CLEA retained more than 60% of the initial activity after six cycles of reuse compared to free enzyme. The high stability and recyclability of CLEA-lipase from CPH make it efficient for different industrial applications.