Engineering a Pseudomonas putida as living quorum quencher for biofilm formation inhibition, benzenes degradation, and environmental risk evaluation.

Bacterial communication interruption based on quorum quenching (QQ) has been proven its potential in biofilm formation inhibition and biofouling control. However, it would be more satisfying if QQ could be combined with the efficient degradation of contaminants in environmental engineering. In this study, we engineered a biofilm of Pseudomonas putida through introducing a QQ synthetic gene, which achieved both biofilm formation inhibition and efficient degradation of benzene series in wastewater. The aiiO gene introduced into the P. putida by heat shock method was highly expressed to produce QQ enzyme to degrade AHL-based signal molecules. The addition of this engineered P. putida reduced the AHLs concentration, quorum sensing gene expression, and connections of the microbial community network in activated sludge and therefore inhibited the biofilm formation. Meanwhile, the sodium benzoate degradation assay indicated an enhanced benzene series removal ability of the engineering bacteria on activated sludge. Besides, we also demonstrated a controllable environmental risk of this engineered bacteria through monitoring its abundance and horizontal gene transfer test. Overall, the results of this study suggest an alternative strategy to solve multiple environmental problems through genetic engineering means and provide support for the application of engineered bacteria in environmental biotechnology.

Can Research Training be Improved in Health Professional Student Curricula? A Qualitative Descriptive Study of Health Students' Experiences with an Integrated Research Training Platform.

Introduction: Project-based learning is currently the status quo for research training for health professional students; however, it alone is not sufficient for holistic development of research skills. One promising style of intervention that can complement project-based research training is a centralised hub of e-learning resources. Therefore, we explored the perception of health professionals in tertiary education, towards the E-learning hub named 'Health and Medical Sciences Research Modules'. Specifically, we explored (1) the role the Modules can play in supporting students in their research training courses, (2) the perception of the selection of topics and content quality and (3) student engagement with content. Methods: Semi-structured interviews were conducted via the online platform Zoom for University of Western Australia 3rd- and 4th-year Dental doctoral students, and 2nd-year Master of Pharmacy students. Interview transcripts were analysed using the framework method, to identify manifest and latent level themes. Results: Eleven participants completed the interview, including five dentistry and six pharmacy students. The analysis yielded numerous manifest level themes including selection and depth of topics, and four latent level themes: content volume and balance, relevance of content to project, alignment and sequencing, and interactivity. Discussion: The present study suggests the Modules content and the online platform were well received as a complementary intervention to project-based learning. However, issues such as content oversaturation emerged as topics which can be addressed to improve the learning experience. These topics should be considered when considering further implementation of e-learning hubs to complement project-based learning, across Australia and worldwide.

Reconstruction and optimization of a Pseudomonas putida-Escherichia coli microbial consortium for mcl-PHA production from lignocellulosic biomass.

The demand for non-petroleum-based, especially biodegradable plastics has been on the rise in the last decades. Medium-chain-length polyhydroxyalkanoate (mcl-PHA) is a biopolymer composed of 6-14 carbon atoms produced from renewable feedstocks and has become the focus of research. In recent years, researchers aimed to overcome the disadvantages of single strains, and artificial microbial consortia have been developed into efficient platforms. In this work, we reconstructed the previously developed microbial consortium composed of engineered Pseudomonas putida KT∆ABZF (p2-a-J) and Escherichia coli ∆4D (ACP-SCLAC). The maximum titer of mcl-PHA reached 3.98 g/L using 10 g/L glucose, 5 g/L octanoic acid as substrates by the engineered P. putida KT∆ABZF (p2-a-J). On the other hand, the maximum synthesis capacity of the engineered E. coli ∆4D (ACP-SCLAC) was enhanced to 3.38 g/L acetic acid and 0.67 g/L free fatty acids (FFAs) using 10 g/L xylose as substrate. Based on the concept of "nutrient supply-detoxification," the engineered E. coli ∆4D (ACP-SCLAC) provided nutrient for the engineered P. putida KT∆ABZF (p2-a-J) and it acted to detoxify the substrates. Through this functional division and rational design of the metabolic pathways, the engineered P. putida-E. coli microbial consortium could produce 1.30 g/L of mcl-PHA from 10 g/L glucose and xylose. Finally, the consortium produced 1.02 g/L of mcl-PHA using lignocellulosic hydrolysate containing 10.50 g/L glucose and 10.21 g/L xylose as the substrate. The consortium developed in this study has good potential for mcl-PHA production and provides a valuable reference for the production of high-value biological products using inexpensive carbon sources.

Engineering Pseudomonas putida To Produce Rhamnolipid Biosurfactants for Promoting Phenanthrene Biodegradation by a Two-Species Microbial Consortium.

Polycyclic aromatic hydrocarbons (PAHs) are a group of organic contaminants that pose a significant environmental hazard. Phenanthrene is one of the model compounds for the study of biodegradation of PAHs. However, the biodegradation of phenanthrene is often limited by its low water solubility and dissolution rate. To overcome this limitation, we engineered a strain of Pseudomonas putida to produce rhamnolipid biosurfactants and thereby promote phenanthrene biodegradation by an engineered strain of Escherichia coli constructed previously in our lab. The E. coli-P. putida two-species consortium exhibited a synergistic effect of these two distinct organisms in degrading phenanthrene, resulting in an increase from 61.15 to 73.86% of the degradation ratio of 100 mg/L phenanthrene within 7 days. After additional optimization of the degradation conditions, the phenanthrene degradation ratio was improved to 85.73%. IMPORTANCE Polycyclic aromatic hydrocarbons (PAHs), which are recalcitrant, carcinogenic, and tend to bioaccumulate, are widespread and persistent environmental pollutants. Based on these characteristics, the U.S. Environmental Protection Agency has listed PAHs as priority contaminants. Although there are many methods to treat PAH pollution, these methods are mostly limited by the poor water solubility of PAHs, which is especially true for the biodegradation process. Recent evidence of PAH-contaminated sites suffering from increasingly severe impact has emerged. As a result, the need to degrade PAHs is becoming urgent. The significance of our study lies in the development of nonpathogenic strains of biosurfactant-producing Pseudomonas aeruginosa for promoting the degradation of phenanthrene by engineered Escherichia coli.