Thanks to all the team members for their tireless efforts, thanks to the instructor's patient guidance, and the groups and individuals who have helped our team. Thanks to everyone who cares about 2019 ECUST. Thanks also to the schools and businesses that have helped our projects. Thanks to teachers especially Pro. Wang Qiyao and Pro. Quan Shu for providing us with experimental guidance and fields. We are grateful to all the staff in East China University of Science and Technology, without whose support , Paper Transformer cannot move forward nor complete.
Our Wet Lab work mainly focused on three branches and one final goal—to construct an engineered E.coli which can turn trash into treasure—it can degrade short plant fibers into bacterial cellulose. To achieve this goal, we planned to work on three different but connected parts.
Miao Zongjie and Jin Qianwei were responsible for the construction and characterization of the inverter and cellubiose operon, Xianglan took charge of the characterization of Bacterial Cellulose synthesis, while Hu Shijia concerned about cellulose decomposition and Liu Xv helped with optimization of the E.coli hemolysin secretion system.
Upon the completion of these three parts, They joined together to give short fibers a brand new look.
Xu Jia, acted as the leader of wiki group, organized all the content uploaded to our wiki and designed some graphics for the wiki. She also self-taught programming and worked on our wiki structure and implementation.
Hu Shijia loves painting and she drew parts of the graphics for our wiki, she was also responsible for reviewing all the text content on our pages.
Yu Shijia, acted as an assistant for Xu and Hu, helping them in all aspects of wiki completion.
Ma Zhenzhen, our Art Design graduate who helped us polish our team logo and designed banner.
2019 ECUST iGEMers wrote all the content for wiki pages together.
Wang Shaohui,acted as the leader of modeling team, he arranged missions and built several models for the paper-transformer program. And he organized on-line discussion to fulfill his duties.
Tan Ningjie is good at searching references about reaction kinetics equations, he acted as an assistant for Wang and completed several steps for each reaction.
Chen Mo was responsible for depicting plots and sheets and she gave some suggestions to correct reaction equations.
Gu Xiaobingmainly investigated the current situation for paper-recycling and designed the whole program of fermentation.
Wu Yuanzhen helped us calculate the profit and benefits of our project and clarified how to make the program better.
Liu Xu, acted as the group leader, she arranged the tasks of human practice and went to several schools ranging from primary to junior schools in August for the popularization of synthetic biology and our project.
Duan Hairong,the principal director of Conference of China iGEMer Community, she made popular science videos, visited communities and scenic spots in Shanghai to investigate and collect public opinions which results has been influencing the orientation of our project.
Xing Yunzhi, planned the practical activities in Shanghai, and he was responsible for visiting Fulun Eco-technology Company. He also conducted questionnaires and collated the information.
Tan Ningjie, was in charge of managing our social media.
They altogether contacted different companies and communicated with other teams.
Also we would like to thank the School of Bioengineering of East China University of Science and Technology for their supports and help.
Advice for Project design
After two months of brainstorm, Paper Transformer was decided as our final project. Initially, we chose mannose as the starting signal of cellulase expression，since there was little mannose in the pulp. At the same time, mannose would be decomposed by seminase, and when mannose was exhausted, microorganisms began to use glucose to produce bacterial cellulose.
Pro. Quan Shu: Can Mannose be trusted?
To verify that our idea is theoretically feasible, we consulted professor Quan Shu, who is well versed in molecular biology and biochemistry. And she told us that Mannose competed with glucose because they share the Phosphoglucose transferase (PTS) system.
This set us thinking more deeply about the feasibility of mannose as a signaling molecule. After consulting relevant manufacturers and the literature, we learned that mannose concentration varied in different pulp, which made the start of the system out of control. Therefore, we chose the common and cheap lactose as signal substance, which also avoided the problem of transport competition.
Our advisors: Choosing a suitable host is important!
We were less experienced in co-culture of the two microorganisms. Furthermore, bacterial cellulose synthase was anchored in the cell membrane of bacteria, so we chose E. coli as our chassis organism. With the suggestion of Mr. Fan, we chose E. coli （DH5α ）as hosts, which was lactose-deficient. Some professors also pointed out that E. coli generally expressed proteins within cells and hardly secreted enzymes, so We tried to solve this problem by using the hemolysin secretion system.
Pro. Wang Qiyao: Note the plasmid compatibility of the dual plasmid system!
We asked professor Wang Qiyao for advice about our ideas. After in-depth discussion, he suggested that since our engineered bacteria need to be responsible for decomposition and synthesis of cellulose respectively, the dual plasmid system could be considered.
Pro. Li Chun: Can the intermediate product accumulate?
In may, we participated in the exchange meeting held by Jiangnan university, during which we consulted professor Li Chun from Beijing Institute of Technology, gaining many valuable suggestions and inspirations. He reminded us that glucose, as a primary carbon source, was difficult to accumulate.
Then we selected cellobiose as the intermediate product, which could not only be accumulated, but also transport across cell membrane to be successfully broken down. This suggestion had a great impact on our project.
Pro. Xia Jianye: Static or dynamic fermentation?
To determine the culture of bacterial cellulose, we sought advice from Xia Jianye who was dedicated to the transformation and optimization of bioreactors. He told us that dynamic fermentation could produce higher yield while static fermentation keep bacterial cellulose from shearing and show better performance.
Thanks to the experiment fields and equipment provided by Prof. Wang Qiyao and Pro. Quan Shu.
Thanks to Prof. Zhang Huizhan for explained to us a lot of expertise on promoters and helped us solve the problem on the cellobiose operator of our duel plasmid system.
Thanks to Pro. Fan Jianhua, Gao Shuhong, Li Youyuan and Li Pengfei for their guidance and patience during weekly sessions.
Thanks to 2108 ECUST iGEM team leader Li Fengting, who not only explicated rules and notes about the iGEM competition, but also selflessly provided guidance and help on experiment details. When we were faced with trouble, he helped us refine the genetic circuits and biobricks, and he shared us principles on plasmid construction and seamless cloning techniques.
Thanks to all the members dedicated to collaboration and got in touch with each other to seek for chances of making our synthetic community stronger. Also we would like to thank Hubei University for their patience and providing several plasmids, which facilitated us to better characterize different secretion system in E.coli.
We express our sincere thanks to all the team members participated in social engagement, who had been continuously contacting with related companies and promoting our project conceptions. We would also like to thank all these institutes along with their warm-hearted staff, who tried their best to reach a hand and inspired us with their accurate knowledge on their specialties. We would like to thank Hangzhou Fulun Eco-Technology Co., Ltd, and its technical consultant Mr. Cao Yandong, who explained the current situation of waste paper recycling to us and indicated that white water was not suitable as the source of fiber for our project, which largely impacted our project design.
Thanks to Chengde Anjing E-Commerce Co., Ltd. for sponsoring our team uniforms.
Thanks to Caixuan 3D Company for their services on hardware 3D printing.
We would also like to express our thanks to the BioScience Department of East China University of Science and Technology for the abundant resources and generous funding.