Team:NWU-CHINA-B/Safety

NWU-CHINA-B

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1. Please upload a photo or two of your lab to the iGEM 2021 server (include your team name in the file name), preferably showing the relevant safety features and paste the link to the file here.



https://static.igem.org/mediawiki/2021/4/44/T--NWU-CHINA-B--C_%29MOSIBIPRTY%24-Z1-L85QK.jpg



2. Describe the goal of your project: what is your engineered organism (or other synthetic biology product, system, or tool) supposed to do? Please include specific technical details and names of important parts.



We further enhanced the activity of glucosidase (SS-BGLY) by producing glucosidase (SS-BGLY) derived from Phyllostachys sulfuris in Pichia pastoris GS115 expression system and optimizing the expression element.



The resulting glucosidase (SS-BGLY) was used to catalyze the conversion of common ginsenosides to rare ginsenosides.



3. Which whole organisms, including viruses and cell lines, are you planning to use (or using) in your project?



Pichia pastoris GS115 competent cell, E.coli Top10 competent cell



4. What risks could these organisms pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab?



Using microbes commonly used in laboratories: Pichia pastoris GS115 and E. coli Top10, the likelihood of infecting humans is very low.



The rate of reproduction in air is very small, and in water environment may cause a large number of reproduction, so the waste solution is treated with high temperature sterilization after the culture.



5. What risks could your chassis pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab?



If E. coli escapes from the laboratory, contaminates the water supply, enters the human gastrointestinal tract, and causes human infections.



If it gets into the soil, it can worsen soil quality and contaminate groundwater and crops.



6. What experiments will you do with your organisms and parts?



In order to catalyze the synthesis of rare ginsenosides, we are going to produce glucosidase derived from pyrifolium sulfide in Pichia coli GS115 substrate cells, and improve the expression and activity of the enzyme through element optimization.



7. What risks could arise from these experiments?



Exposure to these bacteria may pose risks, such as illness caused by E. coli.



In other cases, insufficient inactivation is released into the environment, resulting in an increased risk of disease in the surrounding population.



The inducer methanol is a kind of flammable and explosive material, which needs to be carefully preserved.



8. What safety, security or ethical risks would be involved with such a use?



Our research is mainly to improve the yield, and the final product will not involve safety issues. It may be involved in the experimental operation in the middle of the process because of temperature issues. If Pichia yeast GS115 escapes from the laboratory and the living environment exceeds 32℃, it is toxic to the protein expression and may lead to cell death.



A high yield Pichia pastoris strain expressing glycosidase SS-BGLY was designed and constructed, which may lead to gene loss due to improper preservation in later stage.



9. If your project were fully developed, could any of your engineered organisms or parts spread autonomously in the environment?



Microbes may reproduce under certain conditions, but become nutritionally deficient over time and eventually die.



The expression of the target gene needs to be induced under specific conditions, so the possibility of high expression is very small.



10. How will experts overseeing your project help to manage any of the risks you identified in this form?



Our instructor is very familiar with microbiological experiment, and will train us, supervise and guide us to conduct the experiment safely, and remind us how to reduce the risk in the experiment as much as possible before the experiment.



Wear gloves, masks and protective clothing during surgery.



All used strains, equipment and glassware were sterilized with pressure steam to eliminate contamination.



After the initial disinfection, the waste is put in yellow plastic bags and transferred to the decontamination staff for centralized transport to the medical waste treatment station.



Minimize the risks posed by exposure to these strains and incomplete inactivation.



11. What rules or guidance cover your work which could help to manage any of the risks you identified in Part 2 of this form (in particular Question 11)?



Pathogenic microorganism laboratory biosafety regulations at http://www.gov.cn/zwgk/2005-05/23/content_256.htmWS233-2017



Biological safety of pathogenic micro-organism laboratory general guidelines for http://www.lascn.net/uploadfiles/hyxw/2017/7/201707311037473060.pdf.



12. How will the rules, training, containment and other procedures and practices help to manage any of the risks you identified in Part 2 of this form (in particular Question 10)?



The series we use are non-pathogenic and non-animal.



In terms of safety training, we set up an online conference on laboratory safety precautions, including the use of instruments, the use of chemical reagents, the use of chassis bacteria, etc., emphasizing the importance of sterilization and the necessity of paying attention to prevent bacteria from escaping in the process of experiments.



13. What else can you tell us about any risks associated with your project, how you are managing them, or your compliance with iGEM’s safety and security rules and policies?



No