Journal of biotechnology最新文献

Macrolactins have attracted considerable attention due to their value and application in medicine and agriculture. However, poor yields severely hinder their broader application in these fields. This study aimed to improve macrolactins production in Bacillus siamensis using a combined atmospheric and room-temperature plasma mutagenesis and a microbial microdroplet culture system. After 25 days of treatment, a desirable strain with macrolactins production 3.0-fold higher than that of the parental strain was successfully selected. The addition of 30 mg/L ZnSO₄ further increased macrolactins production to 503 ± 37.6 μg/mL, representing a 30.9 % improvement in production compared to controls. Based on transcriptome analysis, the synthesis pathways of amino acids, fengycin, and surfactin were found to be downregulated in IMD4036. Further fermentation experiments confirmed that inhibition of the comparative fengycin synthesis pathway was potentially driving the increased production of macrolactins. The strategies and possible mechanisms detailed in this study can provide insight into enhancing the production of other secondary metabolites toxic to the producer strains.

Cultivations of Chinese Hamster Ovary (CHO) cells in a perfusion setup were conducted in the presence of super physiological concentrations of L-Arginine to investigate the impact on transmission through the perfusion filter for production of a recombinant domain antibody. Our study revealed that the presence of L-Arginine within the range of 30–50 mM had a positive impact on transmission. However, the higher concentrations were found to have a negative correlation with cell viability, and an optimal concentration of approximately 40 mM was identified. The supplementation of L-Arginine improved overall cultivation performance and enhanced product quality attributes. As a result, our findings demonstrate that the supplementation of L-Arginine to mammalian perfusion cultivations stands as an effective method to address transmission issues, exerting a broad impact on process and production of recombinant proteins.

In this study, a novel electrochemical biosensor was developed for the sensitive and selective detection of the Acinetobacter baumannii gene sequence. The biosensor was created by immobilizing a capture probe specific to the A. baumannii gene on the surface of chitosan-gold modified pencil graphite electrodes. Following solid-state hybridization on the Chit-Au/PGE surface, the target DNA sequence of the A. baumannii was detected by measuring the guanine signal using square wave voltammetry (SWV). All experimental parameters impacting sensor response are examined in order to improve hybridization efficacy, and the electrochemical biosensor's performance. The limit of detection (LOD) for the A. baumannii gene sequence was calculated and found to be 1.93 nM. Three different non-complementary DNA sequences were used to evaluate the assay selectivity, but no interference effect was obtained. Additionally, the potential applicability of the biosensor to real samples was tested in artificial serum media. The suggested electrochemical test procedure is simple, approachable, and quick, making it a convenient approach for the screening of DNA sequence.

Lignin can affect the enzymatic hydrolysis efficiency of lignocellulose. In this study, the lignin isolated from sugarcane bagasse (SCB) pretreated with p-toluenesulfonic acid (PL) was firstly aminated, and then the effects of PL and aminated PL (APL) on the bagasse enzymatic hydrolysis efficiency (EHE) were investigated. The results showed that the addition of PL and APL promoted the EHE, and EHE with APL (73.82 %) was higher than PL (51.39 %). To explore the reason, the data were further analyzed including cellulase adsorption capacity, enzyme activity, cellulase-lignin interaction, and molecular docking. It was found that APL adsorbed more cellulase (27.83 mg protein/g lignin) than PL (4.96 mg protein/g lignin), resulting from the greater interaction force and lower binding free energy between APL and cellulase. The addition of APL more remarkably enhanced the cellobiohydrolase and endoglucanase activities than PL due to more effectively inducing cellulase conformation optimization.

The development of strategies that can permit to adjust the size specificity of immobilized proteases by the generation of steric hindrances may enlarge its applicability. Using as a model ficin immobilized on glyoxyl agarose, two strategies were assayed to generate tailor made steric hindrances. First, ficin has been coimmobilized on supports coated with large proteins (hemoglobin or bovine serum albumin (BSA)). While coimmobilization of ficin with BSA presented no effect on the activity versus any of the assayed substrates, coimmobilization with hemoglobin permitted to improve the immobilized ficin specificity for casein versus hemoglobin, but still significant activity versus hemoglobin remained. Second, aldehyde-dextran has been employed to modify the immobilized ficin, trying to generate steric hindrances to avoid the entry of large proteins (hemoglobin) while enabling the entry of small ones (casein). This also increased the size specificity of ficin, but still did not suppress the activity versus hemoglobin. The combination of both strategies and the use of 37ºC during the proteolysis enabled to almost fully nullify the hydrolytic activity versus hemoglobin while preserving a high percentage of the activity versus casein. The modifications improved enzyme stability and the biocatalyst could be reused for 5 cycles without alteration of its properties.

Selenium (Se) is an essential trace element for life. Seleno-methylselenocysteine (SeMCys) can serve as a Se supplement with anticarcinogenic activity and can improve cognitive deficits. We engineered Escherichia coli for microbial production of SeMCys. The genes involved in the synthesis of SeMCys were divided into three modules–the selenocysteine (SeCys) synthesis, methyl donor synthesis and SMT modules–and expressed in plasmids with different copy numbers. The higher copy number of the SeCys synthesis module facilitated SeMCys production. The major routes for SeCys degradation were then modified. Deletion of the cysteine desulfurase gene csdA or sufS improved SeMCys production the most, and the strain that knocked out both genes doubled SeMCys production. The addition of serine in the mid-logarithmic growth phase significantly improved SeMCys synthesis. When the serine synthetic pathway was enhanced, SeMCys production increased by 12.5 %. Fed-batch culture for sodium selenite supplementation in the early stationary phase improved SeMCys production to 3.715 mg/L. This is the first report of the metabolic engineering of E. coli for the production of SeMCys and provide information on Se metabolism.