Immobilization of laccase on magnetic mesoporous silica as a recoverable biocatalyst for the efficient degradation of benzo[a]pyrene.

Laccase is an efficient green biocatalyst, widely used for the degradation of various organic pollutants. However, free laccase is unstable and difficult to recover, which limits its practical application. In this study, a multilayer core-shell magnetic mesoporous silica (Fe3O4@d-SiO2@p-SiO2) microsphere with high specific surface area (275 m2 g-1) was fabricated for immobilization of laccase. The unique structure of Fe3O4@d-SiO2@p-SiO2 enabled the successful immobilization of laccase. Under the optimal immobilization conditions of laccase concentration of 1.5 mg mL-1, immobilization time of 6 h, immobilization pH of 6, the loading capacity of laccase was up to 567 mg g-1. Compared with free laccase, immobilized laccase exhibited remarkable pH stability, thermal stability and storage stability. Moreover, the immobilized laccase was easy to achieve magnetic recovery and possessed excellent reusability, with its activity remaining 58.2% after 10 consecutive reuses. More importantly, immobilized laccase had good degradation performance for benzo[a]pyrene (BaP), which can achieve rapid and efficient degradation of low concentration BaP over a wide range of pH and temperature. The removal efficiency of BaP was up to 99.0% within 1 h, and still exceeded 35.0% after 5 cycles. The removal of BaP by immobilized laccase was achieved through both adsorption and degradation. The degradation products and possible degradation pathways were determined by GC-MS analysis. This study indicated that Fe3O4@d-SiO2@p-SiO2 could effectively enhance the stability and biocatalytic activity of laccase, which is expected to provide a new clean biotechnology for the remediation of BaP contaminated sites.

Implementation of an interactive mobile application to pilot a rapid assay to detect HIV drug resistance mutations in Kenya.

Usability is an overlooked aspect of implementing lab-based assays, particularly novel assays in low-resource-settings. Esoteric instructions can lead to irreproducible test results and patient harm. To address these issues, we developed a software application based on "Aquarium", a laboratory-operating system run on a computer tablet that provides step-by-step digital interactive instructions, protocol management, and sample tracking. Aquarium was paired with a near point-of-care HIV drug resistance test, "OLA-Simple", that detects mutations associated with virologic failure. In this observational study we evaluated the performance of Aquarium in guiding untrained users through the multi-step laboratory protocol with little supervision. To evaluate the training by Aquarium software we conducted a feasibility study in a laboratory at Coptic Hope Center in Nairobi, Kenya. Twelve volunteers who were unfamiliar with the kit performed the test on blinded samples (2 blood specimens; 5 codons/sample). Steps guided by Aquarium included: CD4+ T-Cell separation, PCR, ligation, detection, and interpretation of test results. Participants filled out a short survey regarding their demographics and experience with the software and kit. None of the laboratory technicians had prior experience performing CD4+ separation and 7/12 had no experience performing laboratory-based molecular assays. 12/12 isolated CD4+ T cells from whole blood with yields comparable to isolations performed by trained personnel. The OLA-Simple workflow was completed by all, with genotyping results interpreted correctly by unaided-eye in 108/120 (90%) and by software in 116/120 (97%) of codons analyzed. In the surveys, participants favorably assessed the use of software guidance. The Aquarium digital instructions enabled first-time users in Kenya to complete the OLA-simple kit workflow with minimal training. Aquarium could increase the accessibility of laboratory assays in low-resource-settings and potentially standardize implementation of clinical laboratory tests.

Aquarium: open-source laboratory software for design, execution and data management.

Automation has been shown to improve the replicability and scalability of biomedical and bioindustrial research. Although the work performed in many labs is repetitive and can be standardized, few academic labs can afford the time and money required to automate their workflows with robotics. We propose that human-in-the-loop automation can fill this critical gap. To this end, we present Aquarium, an open-source, web-based software application that integrates experimental design, inventory management, protocol execution and data capture. We provide a high-level view of how researchers can install Aquarium and use it in their own labs. We discuss the impacts of the Aquarium on working practices, use in biofoundries and opportunities it affords for collaboration and education in life science laboratory research and manufacture.

Synthetic Bistability and Differentiation in Yeast.

Engineered systems that control cellular differentiation and pattern formation are essential for applications like tissue engineering, biomaterial fabrication, and synthetic ecosystems. Synthetic circuits that can take on multiple states have been made to engineer multicellular systems. However, how to use these states to drive interesting cellular behavior remains challenging. Here, we present a cellular differentiation program involving a novel synthetic bistable switch coupled to an antibiotic resistance gene that affects growth in yeast ( S. cerevisiae). The switch is composed of a positive feedback loop involving a novel transcription factor and can be switched ON and OFF via two different transient inducer inputs. By further coupling the bistable switch with an antibiotic resistance gene, we obtained a growth differentiation circuit, where yeast cells can be switched to stable HIGH or LOW growth rate states via transient inducer inputs. This work demonstrates a rationally designed and experimentally validated cellular differentiation behavior in yeast.