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ABSTRACT Penicillin-binding proteins (PBPs) play critical roles in cell wall construction, cell shape maintenance, and bacterial replication. Bacteria maintain a diversity of PBPs, indicating that despite their apparent functional redundancy, there is differentiation across the PBP family. Apparently-redundant proteins can be important for enabling an organism to cope with environmental stressors. In this study, we evaluated the consequence of environmental pH on PBP enzymatic activity inBacillus subtilis. Our data show that a subset of PBPs inB. subtilischange activity levels during alkaline shock and that one PBP isoform is rapidly modified to generate a smaller protein (i.e., PBP1a to PBP1b). Our results indicate that a subset of the PBPs are favored for growth under alkaline conditions, while others are readily dispensable. Indeed, we found that this phenomenon could also be observed inStreptococcus pneumoniae, implying that it may be generalizable across additional bacterial species and further emphasizing the evolutionary benefit of maintaining many, seemingly-redundant periplasmic enzymes. IMPORTANCEMicrobes adapt to ever-changing environments and thrive over a vast range of conditions. While bacterial genomes are relatively small, significant portions encode for “redundant” functions. Apparent redundancy is especially pervasive in bacterial proteins that reside outside of the inner membrane. While conditions within the cytoplasm are carefully controlled, those of the periplasmic space are largely determined by the cell’s exterior environment. As a result, proteins within this environmentally exposed region must be capable of functioning under a vast array of conditions, and/or there must be several similar proteins that have evolved to function under a variety of conditions. This study examines the activity of a class of enzymes that is essential in cell wall construction to determine if individual proteins might be adapted for activity under particular growth conditions. Our results indicate that a subset of these proteins are preferred for growth under alkaline conditions, while others are readily dispensable. 

The use of engineered nanomaterials, defined as those smaller than 100 nm, in the health, energy, agricultural, and environmental sectors is expanding rapidly. As such, human and environmental exposure to these materials is increasing every day. For example, metal-based nanomaterials, such as nanosilver, have become ubiquitous in antibacterial applications ranging from socks and baby bottles to healthcare materials, such as oral fillings. Engineered nanomaterials are also used as antibacterial agents and adjuvants to improve antibiotic delivery or efficacy. However, even nanomaterials that were not designed to be antimicrobial can possess potent bactericidal activity. Alarmingly, there are clear connections between nanomaterial exposure, metal resistance, and antibiotic resistance and it is crucial that we dramatically improve our understanding of both the toxicity of these materials and their ability to permanently change the organisms that they encounter. Emerging research indicates that microbes are capable of adapting to nanomaterial toxicity, often with the same generalizable mechanisms used to overcome antibiotic toxicity. In this perspective, we highlight existing knowledge about microbial response to engineered nanomaterials and the key outstanding questions that must be addressed. 

Video games and immersive, narrative experiences are often called upon to help students understand difficult scientific concepts, such as sense of scale. However, the development of educational video games requires expertise and, frequently, a sizable budget. Here, we report on the use of an interactive text-style video game, NanoAdventure, to communicate about sense of scale and nanotechnology to the public. NanoAdventure was developed on an open-source, free-to-use platform with simple coding and enhanced with free or low-cost assets. NanoAdventure was launched in three languages (English, Spanish, Chinese) and compared to textbook-style and blog-style control texts in a randomized study. Participants answered questions on their knowledge of nanotechnology and their attitudes toward nanotechnology before and after reading one randomly assigned text (textbook, blog, or NanoAdventure game). Our results demonstrate that interactive fiction is effective in communicating about sense of scale and nanotechnology as well as the relevance of nanotechnology to a general public. NanoAdventure was found to be the most “fun” and easy to read of all text styles by participants in a randomized trial. Here, we make the case for interactive “Choose Your Own Adventure” style games as another effective tool among educational game models for chemistry and science communication. 

As lithium intercalating complex metal oxides become more widely used in energy storage devices, there is an increasing need to understand their environmental impact at the end of their life cycle due to the lack of recycling options. In this study, we examine the biological impact of a panel of nanoscale lithium nickel manganese cobalt oxides (LixNiyMnzCo1−y−zO2, 0 < x, y, z < 1, abbreviated as NMCs), to a model Gram-positive bacterium, Bacillus subtilis in terms of cellular respiration and growth. A highly sensitive single-cell gel electrophoresis method is also applied for the first time to understand the genotoxicity of this nanomaterial to bacterial cells. Results from these assays indicate that the free Ni and Co ions released from the incongruent dissolution of the NMC material in B. subtilis growth media induced both hindered growth and cellular respiration. More remarkably, the DNA damage induced by the combination of the two ions in solution is comparble to that induced by the NMC material, which suggests the free Ni and Co ions are responsible for the toxicity observed. A material redesign by enriching Mn is also presented. The combined approaches of evaluating impact on bacterial growth, respiration, DNA damage at a single-cell level, as well as other phenotypical changes allows us to probe the nanomaterial and bacterial cells from a mechanistic prospective, and provides a useful means to an understanding of bacterial response to new potential environmental stressors. 

Engineered nanoparticles are incorporated into numerous emerging technologies because of their unique physical and chemical properties. Many of these properties facilitate novel interactions, including both intentional and accidental effects on biological systems. Silver-containing particles are widely used as antimicrobial agents and recent evidence indicates that bacteria rapidly become resistant to these nanoparticles. Much less studied is the chronic exposure of bacteria to particles that were not designed to interact with microorganisms. For example, previous work has demonstrated that the lithium intercalated battery cathode nanosheet, nickel manganese cobalt oxide (NMC), is cytotoxic and causes a significant delay in growth of Shewanella oneidensis MR-1 upon acute exposure. Here, we report that S. oneidensis MR-1 rapidly adapts to chronic NMC exposure and is subsequently able to survive in much higher concentrations of these particles, providing the first evidence of permanent bacterial resistance following exposure to nanoparticles that were not intended as antibacterial agents. We also found that when NMC-adapted bacteria were subjected to only the metal ions released from this material, their specific growth rates were higher than when exposed to the nanoparticle. As such, we provide here the first demonstration of bacterial resistance to complex metal oxide nanoparticles with an adaptation mechanism that cannot be fully explained by multi-metal adaptation. Importantly, this adaptation persists even after the organism has been grown in pristine media for multiple generations, indicating that S. oneidensis MR-1 has developed permanent resistance to NMC.