Design of Low-Cost Ethanol Production Medium from Syngas: An Optimization of Trace Metals for Clostridium ljungdahlii

Syngas fermentation via the Wood-Ljungdahl (WL) pathway is a promising approach for converting gaseous pollutants (CO and CO2) into high-value commodities. Because the WL involves several enzymes with trace metal components, it requires an adequate supply of micronutrients in the fermentation medium for targeted bioprocessing such as bioethanol production. Plackett-Burman statistical analysis was performed to examine the most efficient trace elements (Ni, Mg, Ca, Mn, Co, Cu, B, W, Zn, Fe, and Mo) and their concentrations for Clostridium ljungdahlii on ethanol production. Overall, 1.5 to 2.5 fold improvement in ethanol production could be achieved with designed trace element concentrations. The effects of tungsten and copper on ethanol and biomass production were determined to be the most significant, respectively. The model developed was statistically significant and has the potential to significantly decrease the cost of trace element solutions by 18–22%. This research demonstrates the critical importance of optimizing the medium for syngas fermentation in terms of product distribution and economic feasibility.

Optimization of Lactic Acid Production from Pineapple By-Products and an Inexpensive Nitrogen Source Using Lactiplantibacillus plantarum strain 4O8

Lactic acid (LA) is used in food, cosmetic, chemical, and pharmaceutical industries and has recently attracted much attention in the production of biodegradable polymers. The expensive substances including carbon and nitrogen sources involved in its fermentative synthesis and the increasing market demand of LA have prompted scientists to look for inexpensive raw materials from which it can be produced. This research was aimed at determining the optimum conditions of lactic acid (LA) production from pineapple by-products and an inexpensive nitrogen source using Lactiplantibacillus plantarum strain 4O8. After collection and preparation of the carbon source (pineapple by-products) and nitrogen sources (by-products from fish, chicken, and beer brewing industries), they were used for the formulation of 4 different media in terms of nitrogen sources. Then, the proximate compositions of promising nitrogen sources were determined. This was followed by the screening of factors (temperature, carbon source, nitrogen source, MgSO4, MnSO4, FeSO4, KH2PO4, and KHPO4) influencing the production of LA using the definitive plan. Lastly, the optimization process was done using the central composite design. The highest LA productions ( 14.64 ± 0.05 g / l and 13.4 ± 0.02 g / l ) were obtained in production medium supplemented with chicken and fish by-products, respectively, making them the most promising sources of nitrogen. The proximate analysis of these nitrogen sources revealed that their protein contents were 83.00 ± 1.41 % DM and 74.00 ± 1.41 % DM for chicken by-products and fish by-products, respectively. Concerning the screening of factors, temperature, nitrogen source, and carbon source were the factors that showed a major impact on LA production in the production medium containing chicken by-products as nitrogen source. A pineapple by-product concentration of 141.75 g/l, a nitrogen source volume of 108.99 ml/l, and a temperature of 30.89°C were recorded as the optimum conditions for LA production. The optimization led to a 2.73-fold increase in LA production when compared with the production medium without nitrogen source. According to these results, chicken by-products are a promising and an inexpensive nitrogen source that can be an alternative to yeast extract in lactic acid production.

Use of Wheat Straw for Value-Added Product Xylanase by Penicillium chrysogenum Strain A3 DSM105774

The present work highlights the valorization of the bulky recalcitrant lignocellulose byproduct wheat straw (WS) for the enhanced production of value-added xylanase by the locally sourced novel Penicillium chrysogenum strain A3 DSM105774 for the first time. The optimized production of xylanase by submerged state of fermentation of WS was achieved using a three-step statistical and sequential approach: one factor at a time (OFAT), Plackett–Burman design (PBD), and Box Behnken design (BBD). Incubation temperature (30 °C), WS, and ammonium sulphate were the key determinants prompting xylanase production; inferred from OFAT. The WS concentration (%(w/v)), yeast extract concentration (%(w/v)), and initial pH of the production medium imposed significant effects (p ≤ 0.05) on the produced xylanase, realized from PBD. The predicted levels of WS concentration, initial pH of the production medium, and yeast extract concentration provoking the ultimate xylanase levels (53.7 U/mL) with an 8.95-fold enhancement, localized by the estimated ridge of the steepest ascent of the ridge analysis path, were 3.8% (w/v), 5.1, and 0.098% (w/v), respectively; 94.7% lab validation. The current data underpin the up-scaling of xylanase production using this eco-friendly, cheap, and robust methodology for the valorization of WS into the value-added product xylanase.

Process Improvisation Strategies for the Enhancement of Lipase Activity

In this present study, lipase-producing bacteria were isolated and screened from an indigenous soil sample and were used for lipase production with high enzyme activity. In the production medium, different production media were screened and lipase production was induced by olive oil, 14 mL/L. It was observed from Luedeking and Piret model that the lipase production was mixed growth associated with maximum activity at 37°C and at pH 7. Statistical optimization using Response Surface Methodology was performed to understand the interaction of different parameters and the standardized conditions obtained were as follows: Peptone 10 g/L, yeast extract 7.5 g/L and olive oil 14 mL/L. The predicted data were validated and the model predicted was significant with a maximum specific activity of 1.1 µmole/min/mg proteins. The lipasespecific activity was enhanced by 10% and 23% after a single parameter and statistical optimization.

Mead Production Using Immobilized Cells of Saccharomyces cerevisiae: Reuse of Sodium Alginate Beads

This work studied the production of mead using second category honey and the immobilized cells of Saccharomyces cerevisiae in sodium alginate, with concentrations of 2% and 4%, and their reuse in five successive fermentations. The immobilized cells with 4% alginate beads were mechanically more stable and able to allow a greater number of reuses, making the process more economical. The fermentation’s consumption of sugars with free cells (control) and immobilized cells showed a similar profile, being completed close to 72 h, while the first use of immobilized cells finished at 96 h. The immobilized cells did not significantly influence some oenological parameters, such as the yield of the consumed sugars/ethanol, the alcohol content, the pH and the total acidity. There was a slight increase in the volatile acidity and a decrease in the production of SO2. The alginate concentrations did not significantly influence either the parameters used to monitor the fermentation process or the characteristics of the mead. Mead fermentations with immobilized cells showed the release of cells into the wort due to the disintegration of the beads, indicating that the matrix used for the yeast’s immobilization should be optimized, considering the mead production medium.

The effect of production conditions on gene expression levels of inulinase of Aspergillus wentii NRLL 1778

In this study, the production of inulinase from Aspergillus wentii, the optimum conditions of that production and how those conditions influence gene expression levels of the enzyme were examined. For inulinase of A. wentii, the time of production was determined as 3 days, the temperature of production as 30°C, the starting pH of the production medium as 6.0, and concentration of Jerusalem artichoke added in to production medium as 3%. When the effect of C and N resources added to growth mediums on inulinase activity was investigated, the highest activity was observed in the medium containing 1% maltose. The medium containing 1% (NH4)2HPO4 was determined to be best growth medium. The enzyme was observed to be stable at pH 5.0-6.0 and to maintain its activity at 50°C for 30 minutes. It was found that gene expression was maximum at 2% Jerusalem artichoke concentration, pH 6.0, 35°C on the 1st day of production. The enzyme gene expression levels were higher compared to other studied resources when 1% cellulose was used as the carbon resource and 1% NH4H2PO4 as the nitrogen resource.

Wild type Bacillus subtilis treated with ethyl methyl sulphonate produced catabolite insensitive mutants with improved cellulase production

The goal of this present investigation was to mutagenize Bacillus subtilis with Ethyl Methyl Sulphonate (EMS), screen the mutants for cellulase production and evaluate the influence of different glucose concentrations on their cellulase production potentials. The wild type B. subtilis was treated with 20, 40, 60 and 80 µl of EMS and the mutants generated were screened for cellulase production in minimal salt medium containing carboxylmethylcellulose (CMC) as the carbon source. Quantitatively, cellulase activity and protein contents were determined by dinitrosalicylic acid and Lowry methods respectively. Seven mutants were developed from each of the EMS concentration bringing the total to twenty-eight from all the concentrations. Approximately 14 and 57% of the mutants developed from 40 and 60µl of EMS had higher cellulase activities than the wild type, while none of the mutants developed from 20 and 80 µl of EMS had better activities than the wild type. The supplementation of 0.2, 0.5, 1.0 and 1.5% glucose in enzyme production medium caused approximately 100, 14, 29 and 14% cellulase repression respectively in the mutants developed from 60µl EMS. Mutants MSSS02 and MSSS05 were considered as catabolite insensitive mutants because their cellulase production were enhanced in comparison to wild type.