Integrated Human Practice

This year our integrated human practice work concentrated on improving the feasibility of our project and exploring its potentials. We contacted experts and technicians to discuss our design and experiment plan with them. Results of these work have been proven helpful to the adjustment of the schematic design and the detailed experiment procedure.

Change of Material -- Interview with Prof. Meng Wenxiang

After the literature research phase, we found microtubule to be an ideal material for our extracellular enzyme loading scaffold for its self-assembly feature and its employing of modifiable subunits. Therefore, we scheduled an interview with professor Meng Wenxiang, an expert on microtubule, before we initiated our experiments.

With Prof. Meng Wenxiang at Institute of Genetics and Developmental Biology

To our disappointment, professor Meng directly pointed out a fundamental flaw in our plan, which was the stability of microtubule could not meet the ideal requirement of the extracellular scaffold. According to his research data, microtubule could not exist in vitro for more than a week even with the help of microtubule-stabilizing drugs, such as taxol^® and its analogues. This defect of microtubule invalidated its potential as the extracellular scaffold material, but it did inspire us to look for a similar but more stable material. So we began our search for materials that met these requirements. Shortly after, we found flagellar filament to be the stable self-assembled biopolymer we wanted. Thanks to professor Meng's advice, we eliminated one possible systematical cause for failure in advance.

Improvement on Experiment Protocol -- Interview with Prof. Zhang Linqi and Her PhD. Students

Since we were new to this technique, and it's imperative for us to enlarge the knowledge base we had on this technique. Hence, we went to Tsinghua University to consult professor Zhang Linqi and her PhD. students, who had tons of experiences and knowledge on this technique.

With PhD. Zhou Panpan at Qinghua University

After our presentation, we received positive feedbacks for professor Zhang and doctor Zhou Panpan. They both considered our project to be promising and would severely increase the capacity of the original system. As for the problems we had encountered in the wet lab, they kindly offered us with practical solutions, helpful advices, and their first-hand experience, which significantly elevated the efficiency of our experiment process.

Modification on Enzyme Choice -- Interview with Director of Research of Beijing Smistyle Sci. & Tech. Development Co., LTD.

According to our research findings, Xynase, an enzyme from cellulase enzyme system, had been applied to substitute the variable D2 domain of flagellin FliC, and the adapted falgellins were successfully assembled into a functioning nano enzyme stick. Then it occurred to us to use the cellulose system to build a prototype to test if our approach can increase the catalytic efficiency. To better define the contrast group for comparison and to better portrait the actual using condition, we contacted Beijing Smistyle Sci. & Tech. Development Co., LTD, whose products covered all three enzymes in the cellulase enzyme system. We made an appointment with Doc. Zhao Lifang, the director of research of Smistyle.

With director of research at Beijing Smistyle Sci. & Tech. Development Co., LTD

Doc. Zhao listened to our power-point presentation attentively, and expressed her sincere appreciation for our project. Nevertheless, she indicated that the application of our improved system on cellulase enzyme system wouldn't fully exhibit the advantages of our improvement. The cost of manufacturing these enzymes had already been optimized to be very low, which made it economically unwise to go through sophisticated recycle procedure for repeated use instead of simply producing a large quantity of them as disposable additions.

At the end of the interview, Doc. Zhao shared her thoughts on more suitable application for our system. Obviously, our target enzyme should be the ones that are hard to produce but highly valuable. This would make the choice of recycle with our system preferable than producing in quantity with a high cost. With these guidelines, we selected PETase, which can hydrolyze PET in low temperature.

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