Team:Hong Kong HKUST/Collaborations

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  Laccase Alkane Metabolism Alkane and MFC (FBA)
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Collaboration

Team HKJS_S

• Date: 29 September 2018

As discussed with Prof. Davis Bookhart, the head of HKUST department of environmental sustainability, for our integrated human practices, a mechanism to reuse or store the CO₂ produced by cellular respiration is required to prevent creating the problem of increasing greenhouse gas in our system.

Team HKJS_S worked to convert carbon dioxide to methane using a modified nitrogenase. By learning more about how this nitrogenase works, we have came to an idea that we can convert the CO₂ produced by cellular respiration of Shewanella into methane that can be used as nutrients for Shewanella under our Alkane metabolism pathway. With this implemented, CO₂ from respiration can be fed back into the system for more electricity generation and therefore increase the production yield.

Apart from the implementation of system, we have also discussed about some of the issues each other have been experiencing during the summer. While the JS_S team successfully clone their constructs using standard assembly and Gibson Assembly, our team have found difficulties in cloning the long sequences we have ordered from IDT into the desired plasmid. With multiple failures in our attempts, we have decided to clone the separate parts using standard assembly and link OmpA to the laccase sequence by adding overhangs to the plasmid using PCR and then perform Gibson Assembly. However, our first attempt of Gibson Assembly had also been unsuccessful and therefore we bring out this issue for further discussion during our meet up.

The JS_S team had helped us to troubleshoot our process on plasmid amplification with overhangs and Gibson Assembly. They suggested us to put the overhang on OmpA instead and linearized the plasmid before any PCR process for higher amplification efficiency and had suggested us on using BL21 E. coli expression strain rather than DH5 alpha strain for greater efficiency in protein production. With the time limit and consideration of our team dynamics, our members with experience in protein extraction had agreed to help the JS_S team with protein extraction using SDS-PAGE. To spare our workload, the JS_S team had also agreed to help us with Gibson Assembly. The result of SDS-PAGE done by our team for collaboration had shown to be successful. Reference can be found from the HKJS_S team’s wiki here

The collaborations mentioned above will contribute to a more detailed characterisation and better product development of both teams.

Team Hong_Kong_JSS

• Date: 2 August 2018

Copper is an essential element to be added to the laccase module to enhance the efficiency of the laccase enzyme. However, copper ions might be added in excess if our MFC are supposed to be maintained by the potential users. This might be toxic to our Shewanella or might even pose hazards to the environment if any accidental release occurs. We therefore aimed to find any collaboration on potential safeguard mechanism to be implemented to our laccase module as a solution to this concern.

Team Hong Kong JSS project focuses on engineering bacteria to absorb accumulated copper in aquaponics and we therefore arranged for a meetup in their school to learn more about each other’s project and discuss the possibilities to integrate their design to help absorb our excess copper. In exchange, we will provide them with modelling guidance.

However, the collaboration is unsuccessful in the end due to our obstacles in cloning for the laccase protein and therefore could not achieve any characterization data to support the needs of absorbing excess copper.

Team SCUT_ChinaA and SCUT_ChinaB

• Date: 27 July 2018

Upon discovering that the SCUT China team B was working on PET plastic degradation, a small meetup was organised by the HKUST team during summer for exchange of ideas in project and experimental design with the 2 teams of SCUT China. Issues such as the ratio of enzyme subunits, surfactant to enhance the attachment of enzymes to the plastic surface for better degradation reaction were discussed. However, we could not identify any possible collaboration on system design and therefore this meetup was just used as a share of experimental insights.

Throughout the summer, we have also attended the following meetups organised by our fellow iGEM teams in attempts for project sharing and discussions :

Virtual conference by Team NYU_Abu_Dhabi

• Date: 15 September 2018

Meetup by team Hong Kong JSS

• Date: 22 September 2018

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