Dataset Information

Sustainable production and preparative purification of thermostable alkaline α-amylase by Bacillus simplex (ON754233) employing natural deep eutectic solvent-based extractive fermentation.

ABSTRACT: Using PEG-based deep eutectic solvents (PDES), the current study proposes extractive fermentation as a sustainable process integration for the production and purification of α-amylase from Bacillus simplex (ON754233). Glucose: PEG 400 outperformed five PDES in terms of tie lie length (58) and slope value (1.23) against sodium sulphatt. Apple cider pomace was used as a low-cost, sustainable carbon source to produce-amylase, with a maximum enzyme production of 2200.13 U/mL. PDES concentration (20% w/v), salt (12.75 w/v), and apple waste (2.75 g/mL) were all optimized using response surface methodology. When scaled upto 3 L benchtop bioreactor, extractive fermentation was proved to be better technology with maximum recovery of 92.4% with highest partition coefficient (3.59). The partially purified enzyme was further purified using a Sephadex G 100 followed by DEAE-Sephadex anion exchange chromatography with a purity fold of 33. The enzyme was found to be thermostable at the temperature (60 °C), remains alkaline (pH 8), and the activity was stimulated in the presence of Mg²⁺ ions. With SDS PAGE electrophoresis, the molecular weight was found to be around 140 kDa. Finally, the enzyme kinetics parameters were evaluated with observed K_m (0.00396 mM) and V_max (37.87 U/mL). Thus scaling up extractive fermentation entails increasing production capacity with improved extraction efficiency using green solvents.

SUBMITTER: Muniasamy R

PROVIDER: S-EPMC10766970 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Sustainable production and preparative purification of thermostable alkaline α-amylase by Bacillus simplex (ON754233) employing natural deep eutectic solvent-based extractive fermentation.

Muniasamy Ramya R Rathnasaamy Senthilkumar S

Scientific reports 20240104 1

Using PEG-based deep eutectic solvents (PDES), the current study proposes extractive fermentation as a sustainable process integration for the production and purification of α-amylase from Bacillus simplex (ON754233). Glucose: PEG 400 outperformed five PDES in terms of tie lie length (58) and slope value (1.23) against sodium sulphatt. Apple cider pomace was used as a low-cost, sustainable carbon source to produce-amylase, with a maximum enzyme production of 2200.13 U/mL. PDES concentration (20% ...[more]

PMID: 38177253

Similar Datasets

Project description:BackgroundBiobutanol is promising and renewable alternative to traditional fossil fuels and could be produced by Clostridium species from lignocellulosic biomass. However, biomass is recalcitrant to be hydrolyzed into fermentable sugars attributed to the densely packed structure by layers of lignin. Development of pretreatment reagents and processes for increasing surface area, removing hemicellulose and lignin, and enhancing the relative content of cellulose is currently an area of great interest. Deep eutectic solvents (DESs), a new class of green solvents, are effective in the pretreatment of lignocellulosic biomass. However, it remains challenging to achieve high titers of total sugars and usually requires combinatorial pretreatment with other reagents. In this study, we aim to develop novel DESs with high application potential in biomass pretreatment and high biocompatibility for biobutanol fermentation.ResultsSeveral DESs with betaine chloride and ethylamine chloride (EaCl) as hydrogen bond acceptors were synthesized. Among them, EaCl:LAC with lactic acid as hydrogen bond donor displayed the best performance in the pretreatment of corncob. Only by single pretreatment with EaCl:LAC, total sugars as high as 53.5 g L-1 could be reached. Consecutive batches for pretreatment of corncob were performed using gradiently decreased cellulase by 5 FPU g-1. At the end of the sixth batch, the concentration and specific yield of total sugars were 58.8 g L-1 and 706 g kg-1 pretreated corncob, saving a total of 50% cellulase. Utilizing hydrolysate as carbon source, butanol titer of 10.4 g L-1 was achieved with butanol yield of 137 g kg-1 pretreated corncob by Clostridium saccharobutylicum DSM13864.ConclusionsEthylamine and lactic acid-based deep eutectic solvent is promising in pretreatment of corncob with high total sugar concentrations and compatible for biobutanol fermentation. This study provides an efficient pretreatment reagent for facilely reducing recalcitrance of lignocellulosic materials and a promising process for biobutanol fermentation from renewable biomass.

Project description:BackgroundTraditional submerged fermentation mainly accumulates intracellular orange pigments with absorption maxima at 470 nm, whereas extractive fermentation of Monascus spp. with Triton X-100 can promote the export of intracellular pigments to extracellular broth, mainly obtaining extracellular yellow pigments with absorption maxima at approximately 410 nm. In this study, a strain of Monascus (M. anka GIM 3.592) that produces high yields of pigments was employed to investigate the differences in pigment fingerprint profiles between submerged and extractive fermentations.ResultsUsing extractive fermentation with this high-yield strain, the extracellular pigments exhibited an absorption maximum at 430 nm, not 410 nm, as previously observed. By comparing the pigment fingerprint profiles between submerged and extractive fermentations, extractive fermentation was found to not only export intracellular pigments to the extracellular broth, but also to form four other yellow pigments (Y1-Y4) that accounted for a large proportion of the extracellular pigments and that were not produced in submerged fermentation. The yields of Y1-Y4 were closely related to the concentration and feeding time point of Triton X-100. Y1-Y4 presented identical UV-Vis spectra with absorption maxima at 430 nm and fluorescence spectra with absorption maxima (emission) at 565 nm. HPLC-MS and the spectral analysis showed that the four pigments (Y1-Y4) had not been previously reported. The results indicated that these pigments may rely on the bioconversion of orange pigments (rubropunctatin and monascorubrin).ConclusionsUsing extractive fermentation with M. anka led to a high yield of extracellular yellow pigments (AU410 nm = 114), and the pigment fingerprint profile significantly differed compared to the results of traditional submerged fermentation. These results provide information and a detailed view of the composition and variation of pigments in extractive fermentation and could also contribute to characterizing pigment metabolism during extractive fermentation.

Dataset Information

Sustainable production and preparative purification of thermostable alkaline α-amylase by Bacillus simplex (ON754233) employing natural deep eutectic solvent-based extractive fermentation.

Publications

Sustainable production and preparative purification of thermostable alkaline α-amylase by Bacillus simplex (ON754233) employing natural deep eutectic solvent-based extractive fermentation.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets