Microbial host engineering for sustainable isobutanol production from renewable resources.

Due to the limited resources and environmental problems associated with fossil fuels, there is a growing interest in utilizing renewable resources for the production of biofuels through microbial fermentation. Isobutanol is a promising biofuel that could potentially replace gasoline. However, its production efficiency is currently limited by the use of naturally isolated microorganisms. These naturally isolated microorganisms often encounter problems such as a limited range of substrates, low tolerance to solvents or inhibitors, feedback inhibition, and an imbalanced redox state. This makes it difficult to improve their production efficiency through traditional process optimization methods. Fortunately, recent advancements in genetic engineering technologies have made it possible to enhance microbial hosts for the increased production of isobutanol from renewable resources. This review provides a summary of the strategies and synthetic biology approaches that have been employed in the past few years to improve naturally isolated or non-natural microbial hosts for the enhanced production of isobutanol by utilizing different renewable resources. Furthermore, it also discusses the challenges that are faced by engineered microbial hosts and presents future perspectives to enhancing isobutanol production. KEY POINTS: • Promising potential of isobutanol to replace gasoline • Engineering of native and non-native microbial host for isobutanol production • Challenges and opportunities for enhanced isobutanol production.

Design and application of a novel culturing chip (cChip) for culturing the uncultured aquatic microorganisms.

Culturing uncultured microorganisms is an important aspect of microbiology. Once cultured, these microorganisms can be a source of useful antibiotics, enzymes etc. In this study, we have designed a novel culturing chip (cChip) to facilitate the growth of uncultured aquatic bacterial community by concentrating the samples. cChip was optimized for microbial growth using known bacteria in the laboratory as a pre-experiment. Then microorganisms from a freshwater lake were concentrated and inoculated, before putting the inoculated cChip in a simulated lake environment and further sub-culturing on laboratory media. High-throughput sequencing and traditional culturing were also performed for comparison. These three methods were able to detect 265 genera of microorganisms in the sample, of which 78.87% were detected by high-throughput sequencing, 30.94% by cChip, while only 6.42% were obtained by traditional culture. Moreover, all microorganisms obtained by traditional culture were also cultured using the cChip. A total of 45 new strains were isolated from the cChip, and their 16S rRNA gene sequences were 91.35% to 98.7% similar to their closest relatives according to NCBI GenBank database. We conclude that the design and simple operation of cChip can improve the culture efficiency of traditional culture by almost 5 times. To the best of our knowledge, this is the first report comparing a novel culturing method with high-throughput sequencing data and traditional culturing of the same samples.

Evaluation of Decontamination Potential of Wet Wipes Against Microbial Contamination of Chinese Spacecraft Materials.

Abstract There are many kinds of microorganisms that inhabit the environment of manned space stations. Wet wipes are a common tool used in space stations to clean and reduce microorganisms on surfaces. Here, we compared the performance of five types of wipes used by the Chinese Space Station (CSS) on orbit before 2021 in terms of microbial decontamination. In previous studies, we found that Bacillus sp. TJ-1-1 and Staphylococcus sp. HN-5 were the most abundant microorganisms in the assembly environment of the CSS. In this study, we used these two bacteria to build different microbial load models to represent the occurrence and non-occurrence of microbial outbreaks in the on-orbit CSS. The results show that the number of microorganisms that can be removed when wiping the surface with high microbial load by wet wipes was higher than that when wiping the surface with low microbial load. For on-orbit daily cleaning and keeping the microbial population within the regulation concentration range, it is suitable to use two pure water wipes per 100 cm2. When the number of microorganisms increases to a degree where astronauts can see the colonies with their naked eyes, the best way to eliminate the problem is to wipe them thoroughly and repeatedly with at least four quaternary ammonium-based wipes every 100 cm2.

Microbial degradation and community structure analysis of hydroxyl-terminated polybutadiene (HTPB).

Hydroxyl-terminated polybutadiene (HTPB) is a curing adhesive that is commonly used in the production of ammunition, and it emerged during the time of war. After entering the peaceful era, several countries around the globe have focused on the destruction of expired ammunition using safe and economical methods in terms of consumption of energy. Microorganisms exhibit a highly efficient and environment friendly degradation capability for variety of refractory substances. Therefore, in this study we screened five strains of microorganisms from five environmental soil samples for their ability to degrade HTPB. These microorganisms were identified as Microbacterium trichothecenolyticum, Microbacterium esteraromaticum, Arthrobacter pascens, Pseudonocardia carboxydivorans and Ochrobactrum anthropic based on 16S rRNA gene similarity index. We observed the uncorroded and corroded HTPB sample through scanning electron microscopy and observed the formation of lot of holes and gullies in HTPB after corrosion. An 18S rRNA gene clone library was constructed for HTPB-degrading fungi. Based on the results of library evaluation, it was found that the structure of the HTPB-degrading fungi community was relatively simple. A total of 54 positive clones were obtained. These clones represented some uncultured microorganisms that were closely related to Scytalidium lignicola, Pseudokahliella and Gonostomum strenuum. This study will help in the implementation of environment friendly degradation strategies for HTPB degradation.

Evaluating Quantitative Measures of Microbial Contamination from China's Spacecraft Materials.

Different methods are used for the quantification of microbial load on spacecrafts. Here, we investigated a number of methodologies currently in use with the intent to identify the most accurate methods for the quantification of microbes on low-biomass metal surfaces such as those used in China's Space Station. In a previous study, we observed a high abundance of Bacillus sp. TJ 1-1 on interior surfaces of China's Space Station, and we therefore undertook this study in which we used a range of 102 to 109 cells/100 cm2 of this strain for setting different contamination levels. Four of the most common analytical approaches (contact plate, spread plate, quantitative PCR, and BacLight™) were used to quantify the number of viable microbial cells associated with the materials of China's Space Station. Results show that, for 102 cells/100 cm2, the contact plate method is the most convenient and reliable. For microbial contamination levels ≥103 cells/100 cm2 and a sampling area of 121 cm2, the BacLight method proved to be most reliable for the detection of live cells. Moreover, a sampling area of 121 cm2 was found to be the most suitable for analysis of metal surfaces for space station interiors, which are usually low in biomass. These results establish suitable sampling and processing methodologies for microbial enumeration of metal surfaces on China's Space Station.

DR5 related autophagy can promote apoptosis in gliomas after irradiation.

As a cancer treatment strategy, irradiation therapy is widely used that can cause DNA breakage and increase free radicals, which leads to different types of cell death. Among them, apoptosis and autophagy are the most important and the most studied cell death processes. Although the exploration of the relationship between apoptosis and autophagy has been a major area of focus, still the molecular mechanisms of autophagy on apoptosis remain unclear. Here, we have revealed that apoptosis was enhanced by the death receptor 5 (DR5) pathway, and the effect of autophagy on apoptosis was promoted by DR5 interacting with LC3B as well as Caspase8 in gliomas after irradiation. Interestingly, we observed that the addition of four different autophagy inducers, rapamycin (RAP), CCI779, ABT737 and temozolomide (TMZ), induced the differences of DR5 expression and cell apoptosis after irradiation. Unlike RAP and CCI779, ABT737 and TMZ were able to increase DR5 expression and further induce cell death. Therefore, we have concluded that DR5 plays a novel and indispensable role in promoting cell apoptosis under irradiation and suggest a potential therapeutic approach for glioblastoma treatment.

The Effect of Gamma-Ray-Induced Central Nervous System Injury on Peripheral Immune Response: An In Vitro and In Vivo Study.

Radiotherapy to treat brain tumors can potentially harm the central nervous system (CNS). The radiation stimulates a series of immune responses in both the CNS as well as peripheral immune system. To date, studies have mostly focused on the changes occurring in the immune response within the CNS. In this study, we investigated the effect of γ-ray-induced CNS injury on the peripheral immune response using a cell co-culture model and a whole-brain irradiation (WBI) rat model. Nerve cells (SH-SY5Y and U87 MG cells) were γ-ray irradiated, then culture media of the irradiated cells (conditioned media) was used to culture immune cells (THP-1 cells or Jurkat cells). Analyses were performed based on the response of immune cells in conditioned media. Sprague-Dawley rats received WBI at different doses, and were fed for one week to one month postirradiation. Spleen and peripheral blood were then isolated and analyzed. We observed that the number of monocytes in peripheral blood, and the level of NK cells and NKT cells in spleen increased after CNS injury. However, the level of T cells in spleen did not change and the level of B cells in the spleen decreased after γ-ray-induced CNS injury. These findings indicate that CNS injury caused by ionizing radiation induces a series of changes in the peripheral immune system.

Antibiotic discovery: combining isolation chip (iChip) technology and co-culture technique.

Antibiotics had been a useful tool for treating bacterial infections since their discovery, but with the passage of time, the evolution of resistance among microbes against antibiotics has rendered them useless. Many approaches are being used to tackle this problem which include discovery of new antibiotics, modification of the existing ones, and elucidating mechanisms of resistance in microbes for a better understanding. In this review, we have discussed that discovery of new antibiotics is a basic need to fight emerging infectious bacteria, and for this purpose, we should target those microbes from the environment which are not easily culturable. For this purpose, culturing technique should be modified to the in situ culturing as nutritional requirements of unculturable bacteria are unknown. Two different cultivation strategies, diffusion chambers and iChip technology, have been reviewed for their excellent improvement in culturing compared to conventional techniques. Since co-culture is also an important factor which can result in exploring new microbial diversity, we hypothesize that if iChip and co-culture can be combined in a single device, it can allow production of novel antibiotics from those bacteria which are difficult to be cultured in the future.