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Laccases (E.C. 1.10.3.2) produced by white-rot fungi (WRF) can be widely used, but the high cost prevents their use in large-scale industrial processes. Finding a solution to the problem could involve laccase production by solid-state fermentation (SSF) simulating the natural growth conditions for WRF. SSF offers several advantages over conventional submerged fermentation (SmF), such as higher efficiency and productivity of the process and pollution reduction. The aim of this review is therefore to provide an overview of the current state of knowledge about the laccase production by WRF under SSF conditions. The focus is on variations in the up-stream process, fermentation and down-stream process and their impact on laccase activity. The variations of up-stream processing involve inoculum preparation, inoculation of the medium and formulation of the propagation and production media. According to the studies, the production process can be shortened to 5–7 days by the selection of a suitable combination of lignocellulosic material and laccase producer without the need for any additional components of the culture medium. Efficient laccase production was achieved by valorisation of wastes as agro-food, municipal wastes or waste generated from wood processing industries. This leads to a reduction of costs and an increase in competitiveness compared to other commonly used methods and/or procedures. There will be significant challenges and opportunities in the future, where SSF could become more efficient and bring the enzyme production to a higher level, especially in new biorefineries, bioreactors and biomolecular/genetic engineering.

The accumulation of polyhydroxyalkanoates (PHAs) by microorganisms usually occurs in response to environmental stress conditions. Therefore, it is advantageous to choose two-step cultivation. The first phase is aimed at maximizing biomass production, and only in the second phase, after setting the suitable conditions, PHA production starts. The aim of this work was to optimize the composition of the minimal propagation medium used for biomass production of Cupriavidus necator DSM 545 using the response surface methodology (RSM). Based on the results from the search for optimization limits, the glucose concentration, the ammonium sulfate concentration and the phosphate buffer molarity were chosen as independent variables. The optimal values were found as follows: the glucose concentration 10.8 g/L; the ammonium sulfate concentration 0.95 g/L; and the phosphate buffer molarity 60.2 mmol/L. The predicted biomass concentration was 4.54 g/L, and the verified value was at 4.84 g/L. Although this work was primarily focused on determining the optimal composition of the propagation medium, we also evaluated the optimal composition of the production medium and found that the optimal glucose concentration was 6.7 g/L; the ammonium sulfate concentration 0.60 g/L; and the phosphate buffer molarity 20 mmol/L. The predicted PHB yield was 54.7% (w/w) of dry biomass, and the verified value was 49.1%.

The viral genome of the SARS-CoV-2 coronavirus, the aetiologic agent of COVID-19, encodes structural, non-structural, and accessory proteins. Most of these components undergo rapid genetic variations, though to a lesser extent the essential viral proteases. Consequently, the protease and/or deubiquitinase activities of the cysteine proteases M and PL became attractive targets for the design of antiviral agents. Here, we develop and evaluate new bis(benzylidene)cyclohexanones (BBC) and identify potential antiviral compounds. Three compounds were found to be effective in reducing the SARS-CoV-2 load, with EC values in the low micromolar concentration range. However, these compounds also exhibited inhibitory activity IC against PL at approximately 10-fold higher micromolar concentrations. Although originally developed as PL inhibitors, the comparison between IC and EC of BBC indicates that the mechanism of their antiviral activity is probably not directly related to inhibition of viral cysteine proteases. In conclusion, our study has identified new potential noncytotoxic antiviral compounds suitable for testing and further improvement.

Polyhydroxyalkanoates (PHAs) represent a promising alternative to commercially used petroleum-based plastics. Pseudomonas oleovorans is a natural producer of medium-chain-length PHA (mcl-PHA) under cultivation conditions with nitrogen limitation and carbon excess. Two-step cultivation appears to be an efficient but more expensive method of PHA production. Therefore, the aim of this work was to prepare a minimal synthetic medium for maximum biomass yield and to optimize selected independent variables by response surface methodology (RSM). The highest biomass yield (1.71 ± 0.04 g/L) was achieved in the optimized medium containing 8.4 g/L glucose, 5.7 g/L sodium ammonium phosphate and 35.4 mM phosphate buffer. Under these conditions, both carbon and nitrogen sources were completely consumed after 48 h of the cultivation and the biomass yield was 1.7-fold higher than in the conventional medium recommended by the literature. This approach demonstrates the possibility of using two-stage PHA cultivation to obtain the maximum amount of biomass and PHA.

Neuraminidase (NA), as an important protein of influenza virus, represents a promising target for the development of new antiviral agents for the treatment and prevention of influenza A and B. Bacterial host strain Escherichia coli BL21 (DE3)pLysS containing the NA gene of the H1N1 influenza virus produced this overexpressed enzyme in the insoluble fraction of cells in the form of inclusion bodies. The aim of this work was to investigate the effect of independent variables (propagation time, isopropyl β-d-1-thiogalactopyranoside (IPTG) concentration and expression time) on NA accumulation in inclusion bodies and to optimize these conditions by response surface methodology (RSM). The maximum yield of NA (112.97 ± 2.82 U/g) was achieved under optimal conditions, namely, a propagation time of 7.72 h, IPTG concentration of 1.82 mM and gene expression time of 7.35 h. This study demonstrated that bacterially expressed NA was enzymatically active.

The accumulation of polyhydroxyalkanoates (PHAs) by microorganisms usually occurs in response to environmental stress conditions. Therefore, it is advantageous to choose two-step cultivation. The first phase is aimed at maximizing biomass production, and only in the second phase, after setting the suitable conditions, PHA production starts. The aim of this work was to optimize the composition of the minimal propagation medium used for biomass production of Cupriavidusnecator DSM 545 using the response surface methodology (RSM). Based on the results from the search for optimization limits, the glucose concentration, the ammonium sulfate concentration and the phosphate buffer molarity were chosen as independent variables. The optimal values were found as follows: the glucose concentration 10.8 g/L; the ammonium sulfate concentration 0.95 g/L; and the phosphate buffer molarity 60.2 mmol/L. The predicted biomass concentration was 4.54 g/L, and the verified value was at 4.84 g/L. Although this work was primarily focused on determining the optimal composition of the propagation medium, we also evaluated the optimal composition of the production medium and found that the optimal glucose concentration was 6.7 g/L; the ammonium sulfate concentration 0.60 g/L; and the phosphate buffer molarity 20 mmol/L. The predicted PHB yield was 54.7% (w/w) of dry biomass, and the verified value was 49.1%.