The fermentation optimization for alkaline protease production by Bacillus subtilis BS-QR-052

IntroductionProteases exhibit a wide range of applications, and among them, alkaline proteases have become a prominent area of research due to their stability in highly alkaline environments. To optimize the production yield and activity of alkaline proteases, researchers are continuously exploring different fermentation conditions and culture medium components.MethodsIn this paper, the fermentation conditions of the alkaline protease (EC 3.4.21.14) production by Bacillus subtilis BS-QR-052 were optimized, and the effect of different nutrition and fermentation conditions was investigated. Based on the single-variable experiments, the Plackett–Burman design was used to explore the significant factors, and then the optimized fermentation conditions, as well as the interaction between these factors, were evaluated by response surface methodology through the Box–Behnken design.Results and discussionThe results showed that 1.03% corn syrup powder, 0.05% MgSO4, 8.02% inoculation volume, 1:1.22 vvm airflow rate, as well as 0.5% corn starch, 0.05% MnSO4, 180 rpm agitation speed, 36°C fermentation temperature, 8.0 initial pH and 96 h incubation time were predicted to be the optimal fermentation conditions. The alkaline protease enzyme activity was estimated to be approximately 1787.91 U/mL, whereas subsequent experimental validation confirmed it reached 1780.03 U/mL, while that of 500 L scale-up fermentation reached 1798.33 U/mL. This study optimized the fermentation conditions for alkaline protease production by B. subtilis through systematic experimental design and data analysis, and the activity of the alkaline protease increased to 300.72% of its original level. The established model for predicting alkaline protease activity was validated, achieving significantly higher levels of enzymatic activity. The findings provide valuable references for further enhancing the yield and activity of alkaline protease, thereby holding substantial practical significance and economic benefits for industrial applications

Six-DOF CFD Simulations of Underwater Vehicle Operating Underwater Turning Maneuvers

Maneuverability, which is closely related to operational performance and safety, is one of the important hydrodynamic properties of an underwater vehicle (UV), and its accurate prediction is essential for preliminary design. The purpose of this study is to analyze the turning ability of a UV while rising or submerging; the computational fluid dynamics (CFD) method was used to numerically predict the six-DOF self-propelled maneuvers of submarine model BB2, including steady turning maneuvers and space spiral maneuvers. In this study, the overset mesh method was used to deal with multi-body motion, the body force method was used to describe the thrust distribution of the propeller at the model scale, and the numerical prediction also included the dynamic deflection of the control planes, where the command was issued by the autopilot. Then, this study used the published model test results of the tank to verify the effectiveness of the CFD prediction of steady turning maneuvers, and the prediction of space spiral maneuvers was carried out on this basis. The numerical results show that the turning motion has a great influence on the depth and pitch attitude of the submarine, and a “stern heavier” phenomenon occurs to a submarine after steering. The underwater turning of a submarine can not only reduce the speed to brake but also limit the dangerous depth. The conclusion is of certain reference significance for submarine emergency maneuvers

Genetic ablation of diabetes-associated gene Ccdc92 reduces obesity and insulin resistance in mice

Summary: Multiple genome-wide association studies (GWAS) have identified specific genetic variants in the coiled-coil domain containing 92 (CCDC92) locus that is associated with obesity and type 2 diabetes in humans. However, the biological function of CCDC92 in obesity and insulin resistance remains to be explored. Utilizing wild-type (WT) and Ccdc92 whole-body knockout (KO) mice, we found that Ccdc92 KO reduced obesity and increased insulin sensitivity under high-fat diet (HFD) conditions. Ccdc92 KO inhibited macrophage infiltration and fibrosis in white adipose tissue (WAT), suggesting Ccdc92 ablation protects against adipose tissue dysfunction. Ccdc92 deletion also increased energy expenditure and further attenuated hepatic steatosis in mice on an HFD. Ccdc92 KO significantly inhibited the inflammatory response and suppressed the NLR Family Pyrin Domain Containing 3 (NLRP3) inflammasome in WAT. Altogether, we demonstrated the critical role of CCDC92 in metabolism, constituting a potential target for treating obesity and insulin resistance

The effect of topological design on the degradation behavior of additively manufactured porous zinc alloy

Abstract The advent of additively manufactured biodegradable porous metals presents a transformative opportunity to meet the criteria of ideal bone substitutes. Precisely tailoring their degradation behavior constitutes a pivotal aspect of this endeavor. In this study, we investigated the effects of topological designs on the degradation profile of laser powder bed fusion (LPBF) Zn scaffolds under dynamic in vitro immersion tests. Specifically, four types of Zn-0.4Mn-0.2Mg scaffolds (beam-based: diamond, face center cubic; surface-based: gyroid, schwarz-P) were designed and fabricated. The degradation mechanism of the scaffolds was comprehensively evaluated using both experimental and simulation methods. The results illuminate the profound impact of structural design on the degradation properties of the Zn alloy scaffolds. The beam-based diamond and face center cubic scaffolds exhibited a degradation rate of 0.08–0.12 mm per year with a relatively uniform degradation mode under dynamic immersion. On the contrary, the surface-based gyroid and Schwarz-P scaffolds demonstrated a notably reduced degradation rate due to lower permeability. This restricted the diffusion of medium ions within the pores, culminating in the accumulation of degradation products and more severe localized degradation. This study underscores the potential of topological design as a compelling strategy for tailoring the degradation profile of additively manufactured biodegradable scaffolds, thereby advancing their suitability as bone substitutes

Erratum to: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (Autophagy, 12, 1, 1-222, 10.1080/15548627.2015.1100356

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Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field