Notebook

Overview

2025.3.6

Team Formation

We established a 9-member team to launch the iGEM project.

2025.3.9 – 2025.3.30

Brainstorming & Project Identification

We conducted multiple brainstorming sessions, identified the project direction centered on astaxanthin synthesis, and finalized the product as an astaxanthin face mask.

2025.3.23 – 2025.4.24

Wet Lab Skill Training & Pathway Investigation

We learned basic molecular biology techniques, investigated the astaxanthin synthesis pathway, and optimized the gene expression model with dual promoters and the idi gene.

2025.4.12 – 2025.8.15

Human Practices & Expert Consultations

We conducted street surveys, interviewed professors and industry experts, attended multiple exchange meetings (South China, iGBA, CCIC), and visited industrial facilities to refine our project design and compliance strategy.

2025.4.10 – 2025.6.20

Dry Lab: Molecular Docking & Mutant Design

We used AlphaFold2, CB-Dock2, PyMOL and other softwares to simulate enzyme-substrate interactions, identified key amino acid sites, and designed mutants to enhance astaxanthin yield.

2025.5.1 – 2025.8.11

Plasmid Construction & Astaxanthin Production

We successfully constructed and verified the pET-ast plasmid, performed transformations, IPTG induction, and HPLC analysis, confirming astaxanthin production and identifying a high-yield mutant HpCrtZ(Cys191Ala).

2025.7.16 – 2025.10.8

Wiki Development & Content Integration

We studied past iGEM projects and collaboratively built and refined the team wiki.

2025.8.14 – 2025.8.23

Product Development: Astaxanthin Face Mask

We developed a gentle olive-oil-based extraction Method, produced astaxanthin-enriched oil, and fabricated the first batch of face mask prototypes.

2025.7 – 2025.10

Presentation Preparation & Mock Defenses

We conducted multiple online and on-site mock defenses, refined presentation skills, and prepared for the iGEM Jamboree.

2025.10.28

iGEM Jamboree

We will present our project at the iGEM Grand Jamboree, showcasing our synthetic biology-based astaxanthin production system and skincare product.

Team Log

Brainstorm

March 9, 2025

Understanding Cellular Factories and Brainstorming

• Objective: Gain an initial understanding of the concept of synthetic biology cell factories and spark project inspiration.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Listened to the professor explain fundamental principles, followed by initial group discussions and brainstorming.
• Outcome: Mastered the fundamental principles of cell factory construction, laying the groundwork for project direction.

March 16, 2025

Project Concept Brainstorming

• Objective: Develop a concrete plan for constructing the cell factory.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Based on previously collected references and information, each member presented their ideas, followed by integrated discussion.
• Outcome: Multiple innovative ideas were consolidated, establishing a preliminary project direction centered on astaxanthin synthesis.

Wetlab

March 23, 2025

Learning Basic Laboratory Operations

• Purpose: Master core foundational techniques for molecular biology experiments.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Learning to use equipment such as pipettes and centrifuges, preparing agarose gels, and performing plasmid extraction and nucleic acid electrophoresis analysis.
• Results: All Participants became familiar with the basic operations of fundamental laboratory techniques and gel electrophoresis technology. We also learned some laboratory safety knowledge.

March 29-30, 2025

Investigating the Astaxanthin Biosynthetic Pathway

• Objective: Determine the optimal biosynthetic pathway for astaxanthin.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Reviewed extensive literature and compared the efficiency of different synthesis pathways.
• Results: An efficient astaxanthin synthesis pathway comprising six key genes and a single T7 promoter was identified, and the corresponding gene sequences were obtained. Plasmid purchase
• Purpose: To make a plasmid contained the astaxanthin synthesis pathway
• Participants: Advisor Ao, Wang Guanquan
• Method: Contacted GenScript Biotech to synthesize the target plasmid.
• Results: The company returned opinions after evaluation.It is too expensive and time-consuming because we want to express genes under one promoter and the entire gene expression module is too large..

Brainstorms

• Purpose: Adjusting gene expression patterns to synthesize astaxanthin-synthesis plasmid
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: discussing and seeking help from professionals
• Results: We believe that asking experts for advice is an excellent idea. Hence, we decide to visit professor Wang two weeks later.

April 3, 2025

Learning Microbial Culture and Laboratory Safety

• Objective: Establish awareness of aseptic techniques and laboratory safety protocols.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Learning the uses and distinctions of various microbial culture vessels, along with biosafety protection knowledge.
• Outcome: The team gained familiarity with standard operating procedures and potential risks in biological laboratories, enabling them to effectively prevent contamination and hazards through proper disinfection and other measures.

April 5, 2025

Simulated Transformation Practice

• Objective: Practice heat-shock transformation and verify reagent viability using a control plasmid.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Performed the transformation procedure using unrelative plasmid and competent E. coli BL21(DE3), including ice-bath, heat-shock, and recovery. Included a no-plasmid control. Spread the mixture on LB plates for overnight incubation.
• Results: The unrelative plasmids plate showed confluent growth, while the control had none, confirming reagent and cell viability. Noted slight inconsistencies in heat-shock timing among members.

April 10, 2025

Practical Plate Pouring and Streaking Techniques

• Objective: Master core skills in microbial isolation and purification.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Learned plate pouring, streaking, spreading techniques, and use of the laminar flow hood, strictly adhering to laboratory safety protocols during practical exercises.
• Results: Team members demonstrated proficient use of petri dishes and laminar flow hoods to complete microbial inoculation and isolation procedures.

April 13, 2025

Brainstorms

• Purpose: Adjusting gene expression patterns to synthesize astaxanthin-synthesis plasmid based on the suggestions given by professor Wang
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: discussing and search online
• Results: We decide take his suggestion as the primary plan.

Reorder plasmid contained astaxanthin synthesis pathway

• Purpose: To obtain a plasmid containing the astaxanthin synthesis pathway
• Participants: Advisor Ao, Wang Guanquan
• Method: Based on the interview with Professor Wang, we developed the astaxanthin synthesis model by adding an additional gene - idi and employing two T7 promoter to express seven genes, which lowered the construction periods and cost. Then we commissioned GenScript Biotech to synthesize the target plasmid.
• Results: Successfully, the company took on the deal, preparing for subsequent validation experiments.

April 17, 2025

Learning Basic Genetic Engineering Techniques

• Objective: To consolidate microbial culture techniques and prepare for subsequent genetic operations.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Reviewed streak plating and spread plating techniques, learned to place inoculated petri dishes in an incubator for cultivation.
• Results: Successfully obtained inoculated microbial culture dishes and mastered the basic procedures for shaking incubator cultivation.

April 20, 2025

Preparation of LB Medium

• Purpose: Verification in the first round of experiments
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian,
• Method:
  1. Turn on the electronic balance, place weighing paper on it, and press "Tabe" after the reading stabilizes.
  2. Weigh 10g sodium chloride, 10g peptone, 5g Yeast extract, and 5g agar powder.
  3. Measure 1L of deionized water using a graduated cylinder. Add to the beaker and mix thoroughly until the solution is clear and transparent with no solid residue.
  4. Dispense the prepared LB medium into 250 mL conical flasks, each containing 100 mL.
  5. Cover with a breathable membrane (rough side up) and secure tightly with a rubber band.
  6. Verify that the water level in the autoclave and the condensation tray meet specifications.
  7. Sterilize the conical flasks containing LB solution in the autoclave at 121°C for 20 minutes.
• Result: Sterile LB medium prepared for inoculation of E. coli.

April 24, 2025

Observation of Microbial Culture Plates

• Purpose: To examine culture Results and reinforce awareness of aseptic techniques.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Observed E. coli colony status after one week of incubation in a constant-temperature incubator to determine contamination presence and reviewed operational procedures.
• Results: Contamination was detected in some petri dishes. The responsible operators reviewed and summarized non-compliant procedures. The unpredicted Results May be caused by unstable winds or hot coating rod is not sufficiently cooled.

Repeated Streak Plate Practice

• Purpose: Reinforce aseptic technique through streak plating to address prior contamination issues
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Performed the transformation procedure using unrelated plasmid and competent E. coli BL21(DE3), including ice-bath, heat-shock, and recovery. Included a no-plasmid control. Spread the mixture on LB plates for overnight incubation.
• Results: Most plates yielded isolated single colonies. Two plates showed contamination, potentially due to insufficient hood stabilization time or improper loop cooling. In Next Steps, we will ensure laminar flow hood runs for >20 mins before use and allow loops to cool completely post-flaming.

May 1, 2025

Purposes of the next four days: The plasmid named pET-ast was synthesized by GenScript Biotech and returned to us. We need to test if it can synthesize astaxanthin as we designed.

Preparation of LB Medium

• Purpose: Verification in the first round of experiments

• Participants: Jia Jingyi, Qin Tian,

• Method:

  1. Turn on the electronic balance, place weighing paper on it, and press "Tabe" after the reading stabilizes.

  2. Weigh 10g sodium chloride, 10g peptone, 5g Yeast extract, and 5g agar powder.

  3. Measure 1L of deionized water using a graduated cylinder. Add to the beaker and mix thoroughly until the solution is clear and transparent with no solid residue.

  4. Dispense the prepared LB medium into 250 mL conical flasks, each containing 100 mL.

  5. Cover with a breathable membrane (rough side up) and secure tightly with a rubber band.

  6. Verify that the water level in the autoclave and the condensation tray meet specifications.

  7. Sterilize the conical flasks containing LB solution in the autoclave at 121°C for 20 minutes.

• Result: Sterile LB medium prepared for inoculation of E. coli.

 

E. coli transformation via heat shock Method

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida

• Method:

  1. Retrieve the plasmid from the -20°C freezer and thaw at room temperature.

  2. Remove frozen E. coli BL21 (DE3) competent cells from -80°C ultra-low temperature freezer, let stand for 5 min. Perform subsequent steps on ice.

  3. In a clean bench, add 1μL of target plasmid to the competent cells. Gently tap the tube containing the cells, return it to ice, and ice-bathe for 30 minutes. Heat-shock at 42°C for 45 seconds, then place back on ice and let stand for 2 minutes.

  4. Transfer to a clean bench. Add 800μL SOC medium, 200 mM MgCl₂, and incubate at 37°C on a shaking incubator for 1 hour.

  5. After recovery, spread onto LB solid plates containing Amp and incubate at 37°C for 16 hours.

• Result: Plasmid transformation completed.

Restriction enzyme digestion

• Participants: Jia Jingyi, Qin Tian
• Method: Prepare the reaction in a 0.2 mL EP tube on ice. After adding all agents, briefly centrifuge the mixture and then incubate at 37°C for 1 hour.

Agent	Amount
Plasmid DNA	1 µg
FastDigest Pac I	1 µL
FastDigest Nde I	1 µL
10 × FastDigest Green Buffer	2 µL
ddH₂O	To final 20 µL

• Result: Two clear and correct bands were observed, which indicated that the plasmid construction was successful.

 

Take photos

• Participants: Wang Xiwu, Wang Yichun
• Method: record our products and the process
• Result: clear and beautiful picture

May 2, 2025

Pick and culture transformants Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian

• Method:

  1. Using clean bench. Turn on the UV light 30 minutes in advance. After turning off the UV light, activate the fan for 20–30 minutes.

  2. Remove overnight plates from the 37°C incubator. Observe whether bacteria have grown sufficiently, whether colonies are isolated, and whether morphology (size, color, edges) is consistent.

  3. Using a pipette tip, gently pick single colonies (3 per plate) within the laminar flow hood.

  4. Transfer the tip into a 14-mL culture tube containing 3.5 mL LB medium (containing Amp). Incubate overnight at 37°C, 220 rpm to obtain the seed culture.

• Results: Numerous monoclonal colonies grew on the transformation plates, and E. coli was amplified via liquid culture.

 

Take photos

• Participants: Wang Xiwu, Wang Yichun
• Method: record our products and the process
• Result: clear and beautiful picture

May 3, 2025

IPTG Induction and Centrifugal Harvesting

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian

• Method:

  1. Inoculate overnight seed culture at a 1:100 ratio into 100 ml LB medium (containing Amp) in a conical flask. Incubate at 30°C, 220 rpm until OD₆₀₀ reaches 0.5–0.6 (approximately 4.5 hours).

  2. Add 100 µL of IPTG (final concentration 0.1 mM) per 100 mL of culture to the conical flask. Induce at 30°C for 5 hours.

  3. Label each 50 mL centrifuge tube and weigh the empty tube (without cap).

  4. Collect the fermentation broth from the flasks into 50 mL centrifuge tubes, 45 mL per tube. Centrifuge at 6000 rpm for 10 min at room temperature.

  5. Discard the supernatant. Resuspend the cell pellet with 45 mL of deionized water. Centrifuge again for 10 min at 6000 rpm. Discard the supernatant and collect the cells.

  6. Repeat the fifth step to collect cells.

• Results: After IPTG induction, the culture colour changed from pale yellow to red, the color change preliminarily indicates the possible expression of the target pathway and astaxanthin synthesis, pending further verification by HPLC

Vacuum Freeze-Drying of E. coli cells

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian

• Method:

  1. Wrap the centrifuge tube opening with plastic wrap, secure tightly with a rubber band, then pierce 5–10 small holes in the wrap using a toothpick.

  2. Immerse the bottom of the tube in liquid nitrogen for 4 minutes to freeze. Bundle three frozen samples together with a rubber band.

  3. Turn on the vacuum freeze-dryer. Place pre-frozen samples onto the sample tray, then insert the entire tray and rack into the cold trap chamber. Cover with the acrylic lid and close the drain and vent valves.

  4. Wait until the temperature drops to -50°C. Turn on the vacuum pump and vacuum gauge. Once the vacuum level reaches 50 Pa, leave the chamber and continue freeze-drying for 16 hours.

• Result: Completely dried E. coli powder obtained, exhibiting a red colour.

Take photos

• Participants: Wang Xiwu, Wang Yichun
• Method: record our products and the process
• Result: clear and beautiful picture

May 4, 2025

Pigment Extraction

• Participants: Wang Guanquan, XU ALAN TSZ HONG

• Method:

  1. Crush the dried bacterial cells on weighing paper using a 50 mL centrifuge tube. Weigh 10–50 mg into a centrifuge tube.

  2. Add 600µL of methanol: isopropanol solution (8:2) and an appropriate amount of glass beads.

  3. Place in a homogenizer and grind for 10 minutes (30 cycles: 15 seconds each, 5-second pause between cycles, at 4°C).

  4. Centrifuge at 12,000×g for 5 min. Observe yellowish-white bacterial cells and red supernatant.

  5. Using a syringe, aspirate all supernatant. Filter through a 0.22μm organic filter membrane (to remove particles larger than 0.22μm and prevent column from clogging). Directly filter the filtrate into a brown centrifuge tube for storage at -20°C.

• Results: The extracted solution containing astaxanthin was obtained and is suitable for HPLC detection.

Carotenoids were detected via high-performance liquid chromatography (HPLC).

• Participants: Wang Guanquan, Liu Lida, Jia Jingyi, Qin Tian

• Method:

  (1) Sample Preparation for Instrument Analysis

  Label samples (Group Number + A/B/C), place them sequentially into the HPLC autosampler, and note their positions.

  (2) Instrument Startup, Parameter Setup, and Analysis

  1. Check if the mobile phase is sufficient; replenish if insufficient. Inspect and replace the chromatographic column as needed.

  2. Turn on the instrument power and launch the chromatography workstation software.

  3. Instrument initialization: Download the target Method to the instrument, initiate self-test, purge the tubing, and flush the column.

  4. Call the appropriate run Method and equilibrate the column.

  5. Edit the sample run sequence list and set parameters.

  6. After column equilibration, run the sample sequence list and initiate sample injection.

  7. Shut down after run completion. When powering off, sequentially turn off components from bottom to top: pumps, detectors, etc., then close the chromatography workstation software.

• Result: Calculate astaxanthin and others’ yields. And we find out so much canthaxanthin that cannot be catalyzed. So we will try to decrease the yield of canthaxanthin in next stage. Hence, we need to visit another experts who can give us great suggestions.

Take photos

• Participants: Wang Xiwu, Wang Yichun
• Method: record our products and the process
• Result: clear and beautiful picture

June 16, 2025

Plasmid Transformation and Creative Plating

• Purpose: Transforming pET-ast plasmid into E. coli BL21(DE3) and create plate drawing.

• Participants: Wang Xiwu, Wang Yichun, Xu Wenxuan

• Method:

  1. Thawed competent cells on ice, added 1μL pET-ast plasmid, and performed heat shock transformation.

  2. Spread the mixture on LB plates and incubated overnight at 37°C.

  3. In clean bench, picked existing colored E. coli colonies and streaked them on LB plates into designed pictures. Incubated overnight at 37°C.

• Results: Transformation plates and culturing until visible bacterial drawing were obtained.

Take photos

• Participants: Wang Xiwu, Wang Yichun
• Method: record our products and the process
• Result: clear and beautiful picture

June 17, 2025

Transformation of pET-ast and selection transformants accumulated astaxanthin

• Purpose: Purify pET-ast transformants and screen for astaxanthin-producing clones.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Qin Tian, Jia Jingyi
• Method: Under aseptic conditions, single colonies were picked and streaked onto LB （ Amp ）plates. Plates were incubated at 37°C in an inverted incubator.
• Results: Transformants successfully grew into single colonies. Orange-red colored colonies indicated successful astaxanthin production.

June 20, 2025

Order mutant plasmid contained HpCrtZ mutants

• Purpose: To obtain a plasmid contained HpCrtZ mutants
• Participants: Advisor Ao, Wang Guanquan
• Method: Based on the dry-lab Results, we obtained four HpCrtZ mutants. Then we synthesized them by GenScript Biotech and replaced the HpCrtZ gene in pET-ast plasmid.
• Results: The company took on the deal, preparing for subsequent validation experiments. We will finally obtain the mutant plasmids including pET-ast (Ser96Ala, Ile102Ala, Cys191Ala, Thr213Ala)..

June 28, 2025

Preliminary Investigation into the Effect of IPTG Concentration on Astaxanthin Production

• Objective: Optimizing the induction conditions (IPTG concentration),.

• Participants: Wang Guanquan, XU ALAN TSZ HONG

• Methods:

  1. Inoculate engineered bacteria containing the pET-ast plasmid into LB medium.

  2. When the OD600 reaches 0.5–0.6, induce with different concentrations of IPTG (e.g., 0.1 mM, 0.5 mM, 1.0 mM), and include a control group without IPTG.

  3. Induce under the same conditions (30°C, 220 rpm) for 5 hours.

  4. Collect the bacterial cells, visually observe the colour intensity, and perform preliminary absorbance measurements of the crude pigment extract (e.g., measure absorbance at 480 nm, where astaxanthin has an absorption peak).

  5. Absorbance measurement: The crude extract from the 0.1 mM group exhibited the highest absorbance value.

• Outcome: Preliminary Results indicate that 0.1 mM is the optimal induction concentration. However, it was also noted that relying solely on colour and crude extract absorbance is not precise. Therefore, in subsequent formal experiments with mutants, 0.1 mM IPTG will be uniformly used, and accurate quantification must be performed using HPLC.

Culturing modified E. coliin large scal.

• Purpose: culturing modified E. coli in large scal to obtain enough E. coli power.
• Participants: Liu Lida, Qin Tian, Jia Jingyi
• Method: pick single colonies containing the astaxanthin synthesis plasmid from the plates. From each plate, select three colonies and inoculate them into 250 mL Erlenmeyer flasks containing 80 mL LB medium. Cultivate overnight at 30 °C with shaking at 220 rpm to obtain seed cultures.
• Results: Seed cultures were obtained from single colonies.

July 5-6, 2025

Preparation of LB Medium

• Purpose: Verification in the first round of experiments
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as April 20,2025
• Result: Sterile LB medium prepared for inoculation of E. coli.

Culturing modified E. coli in large scale

• Purpose: culturing modified E. coli in large scal to obtain enough E. coli power.
• Participants: Liu Lida, Qin Tian, Jia Jingyi
• Method:inoculate the seed culture into glass Erlenmeyer flasks containing 800 mL LB medium (Amp) at a ratio of 1:100. Incubate at 30 °C with shaking at 220 rpm until the OD600 reaches 0.6, then add IPTG to a final concentration of 0.1 mM. Induce at 30 °C for 5 h, and harvest the cells by centrifugation at 8000 rpm for 8 min at 4 °C. The collected bacteria cells were subsequently freeze-dried in vacuum to obtain power, followed by purification and collection.
• Results: Freeze-dried bacterial biomass containing the astaxanthin were obtained.

July 17, 2025

Purposes of the next four days: The new plasmid was synthesized by GenScript Biotech and returned to us. We need to test if it improve the productivity of astaxanthin.

Verification of synthetic plasmid contained HpCrtZ mutants

• Purpose: Determine mutation sites through sequence alignment
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Blast the sequence by SnapGene software
• Results: The mutant plasmid pET-ast (Ser96Ala, Ile102Ala, Cys191Ala, Thr213Ala) was synthesized by GenScript Biotech. We confirmed the mutant sites. We obtained and propare for subsequent validation.

E. coli transformation via heat shock Method

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as May 1, 2025
• Result: Plasmid transformation completed.

July 18, 2025

Pick and culture transformants

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as May 2, 2025
• Results: Numerous monoclonal colonies grew on the transformation plates, and E. coli was amplified through liquid culture.

July 19, 2025

E. coli IPTG Induction and Centrifugal Harvesting

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as May 3, 2025
• Results: After IPTG induction, the culture color changed from pale yellow to red, indicating successful expression of the target gene and astaxanthin synthesis.

 

Vacuum Freeze-Drying of E. coli

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as May 3, 2025
• Result: Completely dried E. coli powder obtained, exhibiting a red color.

July 20, 2025

Pigment Extraction

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as May 4, 2025
• Results: The extracted solution containing astaxanthin was obtained and is suitable for HPLC detection.

Carotenoids were detected via high-performance liquid chromatography (HPLC).

• Participants: Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: the same as May 4, 2025
• Result: HPLC analysis indicated that the astaxanthin yield in the mutant strain (Cys191Ala) was approximately 1.28-fold higher compared to the original strain under the tested conditions. Following the successful verification of high-yield astaxanthin production by our engineered strains, subsequent R&D efforts were directed toward product development.

August 10, 2025

Large-Scale Cultivation of E. coli transformant contained pET-ast (Cys191Ala)

• Purpose: To meet the raw material demand for face mask production, we performed an 800 mL scale-up cultivation using pET-ast (Cys191Ala) plasmid.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Qin Tian
• Method inoculate the seed culture into glass Erlenmeyer flasks containing 800 mL LB medium (Amp) at a ratio of 1:100. Incubate at 30 °C with shaking at 220 rpm until the OD600 reaches 0.6, then add IPTG to a final concentration of 0.1 mM. Induce at 30 °C for 5 h, and harvest the cells by centrifugation at 8000 rpm for 8 min at 4 °C. The collected bacteria cells were subsequently freeze-dried in vacuum to obtain power, followed by purification and collection.
• Result: A substantial amount of red, astaxanthin-rich biomass was successfully obtained. The astaxanthin-rich power would be reserved as raw material for face mask production.

August 11, 2025

Pigment Extraction

• Participants: Jia Jingyi, Qin Tian
• Method: the same as May 4, 2025
• Results: The extracted solution containing astaxanthin was obtained and is suitable for HPLC detection.

Carotenoids were detected via high-performance liquid chromatography (HPLC).

• Participants: Jia Jingyi, Qin Tian
• Method: the same as May 4, 2025
• Result: HPLC analysis indicated that the astaxanthin yield was great that could be used to make astaxanthin-oil.

Dry lab

April 10, 2025

Learn to search for genes and proteins through the NCBI website.

• Purpose: Learn to search for genes and proteins through the NCBI website.
• Objective: Search for gene information related to astaxanthin synthesis using NCBI..
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Open the NCBI website, select the data type in the search bar, and filter the sequence information that meets the requirements from the search Results.
• Results: The gene expression sequences related to the synthesis of astaxanthin have been confirmed.

April 13, 2025

Learn how to use SnapGene

• Purpose: Learn how to use SnapGene
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Download and install SnapGene. Use the software to analyze the protein sequences of genes related to astaxanthin synthesis, such as HpCrtZ.
• Results: Obtain the amino acid sequences of the genes related to astaxanthin synthesis.

April 17, 2025

Learn to use the AlphaFold 2 software

• Purpose: Using AlphaFold 2 to analyze the protein structures of genes related to astaxanthin synthesis.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Open the official website of AlphaFold 2 database, search for the target protein, and download the three-dimensional structure file of the enzyme..
• Results: Got a more comprehensive understanding of the structure of proteins related to astaxanthin synthesis

June 8, 2025

Study enzyme-substrate docking simulation

• Objective: Master molecular docking between enzymes and substrate molecules

• Method: Structure Acquisition: Obtain the initial structure of the enzyme protein via AlphaFold2 (ID: Q9PK6); download .sdf structure files for substrates (β-carotene/Canthaxanthin) from PubChem.

  1. Binding site prediction: Perform binding pocket analysis on the enzyme protein using PrankWeb.

  2. Molecular Docking: Simulate substrate-enzyme protein docking using CB-Dock2 to obtain binding energy scores.

  3. Site-Specific Mutation: Employ PyMOL with the PySwissSidechain plugin to mutate specified sites (S96, I102, C191, T213) to alanine (Ala) and save the new structure.

  4. Validation Analysis: Redock the mutated enzyme structure with the substrate to compare binding energy changes.

• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian

• Results: Successfully obtained mutant protein models and completed preliminary docking simulations with the substrate, yielding binding energy data for wild-type versus mutant comparison. Molecular docking simulations revealed reduced binding energy between the mutant and substrate, algorithmically validating its potential for higher yield.

June 13, 2025

Identification of the β-Carotene Hydroxylase Pocket Region

• Objective: Identify the active site pocket of β-carotene hydroxylase
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Jia Jingyi
• Method: Based on last week's dry lab operations, report on β-carotene hydroxylase protein structure, pocket site amino acids, and key amino acid sites involved in substrate binding (outline the general approach)
• Results: Identified amino acid sites within the β-carotene hydroxylase pocket region. Amino acids interacting with Canthaxanthin include: TYR88 TRP152 HIS165 GLU174 ASN176 ASP177 PHE179 ALA180 ASN183 GLY216 TYR219 MET220 HIS223 ASP224 ARG230; Amino acids involved in β-carotene docking include: TYR88 VAL99 ILE102 ALA103 PHE105 ALA106 LEU109 MET140 TRP152 ASN176 ASP177 PHE179 ALA180 ASN183 GLY184 ALA187 MET188 CYS191 THR192 PHE195 TRP196 LE U210 ILE212 THR213 GLY216 MET217 TYR219 MET220 HIS223 ASP224.

June 19, 2025

Identification of Key Amino Acid Sites

• Objective: Identify critical amino acid residues through alanine scanning
• Participants: Wang Guanquan, Wang Xiwu
• Method:

  1. Describe the protein structural features of carotene hydroxylase (e.g., number of alpha helices, beta folds)

  2. Describe the pocket region of carotene hydroxylase and the amino acid sites involved in the pocket region

  3. Perform molecular docking simulations between carotene hydroxylase and two substrate molecules

  4. Mutate key amino acid sites to alanine, then re-simulate docking to compare changes in binding energy relative to substrate interaction

• Results: After mutation, four key amino acid sites were identified. Their substitution with alanine reduced binding energy with Canthaxanthin, enhancing substrate binding and improving catalytic efficiency.

June 20, 2025

Discussion on validating key amino acid sites

• Objective: Experimental validation of amino acid mutation sites
• Participants: Wang Xiwu, Wang Guanquan,
• Method: To analyze binding energy changes after alanine mutations at each amino acid site within the β-carotene hydroxylase pocket region, obtain mutated sequences via gene synthesis and replace the β-carotene hydroxylase (HpCrtZ) in pET-ast.
• Results: Four amino acid mutation sites were identified including Ile102Ala, Ser96Ala, Cys191Ala, and Thr213Ala. To systematically evaluate the effect of each site, we decided to construct four separate mutant plasmids. Each plasmid will contain a single alanine substitution (e.g., S96A, I102A, etc.) at one of these sites, replacing the wild-type HpCrtZ gene in the original pET-ast backbone. Alanine mutants were chemically synthesized by GenScript Biotech Corporation (Nanjing). The synthesized genes replaced wild-type β-carotene hydroxylase (HpCrtZ) on pET-ast. This approach allows us to pinpoint the contribution of each individual mutation. If the productivity of certain mutants decreases, we can focus on the beneficial ones and explore combinations in the future.

Human practices

April 12, 2025

Street investigation

• Purpose: We conduct offline street surveys to directly know common people's views on astaxanthin and skincare products, and gather suggestions on anti-aging ones.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun
• Method: We asked Shenzhen citizens three core questions: 1. Do you purchase anti-aging skincare products because you care about wrinkles on your face? 2. Are you familiar with astaxanthin and its effects? 3. Are you willing to accept skincare products made from astaxanthin produced by synthetic biology techniques?
• Outcome: the survey Results revealed a pattern: Compared with male respondents, more female respondents would choose to buy anti-aging products due to wrinkles. Citizens who are willing to buy products know relatively little about astaxanthin, but most of them have a demand for skincare yet are restricted by price. They are willing to try astaxanthin skincare products produced by synthetic biology techniques that comply with market regulations.

April 12, 2025

Interview professor Wang Chaogang

• Purpose: Selecting cell factory chassis requires considering factors like genetic operability. We consulted Prof. Wang on using E. coli for producing astaxanthin, seeking inspiration as our approach underperformed.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan
• Method: We consulted Professor Wang on cell factory design. He mentioned key aspects, we discussed chassis choice (he confirmed E. coli with its advantages), astaxanthin synthesis model optimization, and pET-Dute1 vector. The most significant advice he gave is use idi to strengthen supplement of primes and use two Promoters.
• Outcome: we decided to use E. coli as the chassis for our project, which could ensure efficient astaxanthin production. The gene idi was intruduced into our synthesis model and a T7-pro dual-promoter was used to optimize gene expression efficiency.

May 5, 2025

Investigate the actors

• Purpose: one of the interviewees, who we interviewed last month, inspire us the made us wonder whether we could use our astaxanthin products to create a red makeup that is more skin-friendly. We wanted to discuss this possibility.
• Participants: Xu Wenxuan, Wang Xiwu
• Method: we arranged this meeting with the actor who was one of the interviewees suffering from allergy of make-up. He told us some actors abandon their career due to allergy.
• Outcome: We decided to take advantage of the red pigment feature of astaxanthin, providing them with a less harmful antioxidant makeup.

May 8, 2025

Simulated Presentation at the South China Exchange Meeting

• Purpose: To familiarize with the PowerPoint presentation and enhance teamwork coordination among members
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian
• Method: Simulated presentation of our PowerPoint, followed by feedback and revision suggestions from faculty members and professors.
• Outcome: We revised deficient sections and enhanced our presentation skills, enabling smoother and more natural delivery of the PowerPoint.

May 10, 2025

SUSTech-SynBio communication

• Purpose: Seven teams took turns to give presentations on aspects such as project design and HP.
• Participants: Xu Wenxuan, Wang Xiwu
• Method: They actively interacted in the Q&A session and had in-depth discussions and exchanges on issues like the details of the experiments, the specific significance of HP work, and the division of labour for HP.
• Outcome: We listened to the report on SZU-China's HP work last year. They did a wonderful job in integrating inspiration and stories. SUSTechMed's planning for HP work was ingenious as they divided the groups into three circles. Inspired by them, we divided our groups into four circles according to the 4e principle and optimized our project plan based on the feedback.

May 15, 2025

Mock Defense for the South China Exchange Conference

• Purpose: To simulate the entire process of the South China Exchange Conference.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian, Zu Yuhan
• Method: A professor acted as the mock judge. We presented our project once, then conducted a mock defense, recording the questions posed.
• Outcome: Each member gained a general understanding of project components outside their primary responsibility.

May 17, 2025

Attended the South China Exchange Meeting

• Purpose: To understand other teams' progress and research directions
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian, Zu Yuhan
• Method: Presented our project, created cultural and creative products, interviewed other teams, and learned about their projects
• Outcome: Successfully promoted our team; refined our proposal based on feedback from other teams; deepened our understanding of our project; documented the experimental projects of competitive teams. The presents we offer to other teams is popular, so we decide to produce the same products in the next time.

May 22, 2025

Summary of issues encountered at the South China Exchange Meeting and other teams' projects

• Purpose: To summarize the Outcomes of the South China Exchange Meeting
• Participants: Wang Xiwu
• Method: Compile all team members' recorded findings.
• Results: We identified no significant differences in progress compared to other teams. Additionally, we reviewed some aspects of other teams that we noticed but were not familiar with, such as formulae.

May 31, 2025

Expert Interview

• Purpose: To gain in-depth knowledge of industrial astaxanthin production and explore feasibility and existing solutions Expert: Professor Wang Jiangxin
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG
• Method: Discussion with the professor
• Outcome: He gave us some suggestions, for example, breaking cell membrane in low temperature to protect biological activity, optimizing genes selection, increasing enzyme activity through mutation.

June 15, 2025

Expert Interview

• Purpose: optimise the operation, particularly IPTG Expert: Professor Cheng Weizhao
• Participants: Xu Wenxuan, Wang Xiwu, Wang Guanquan, XU ALAN TSZ HONG
• Method: Discussion with the professor, in which parts we can optimise
• Outcome: we decided to operate a research to find out which concentration of IPTG can increase yield.

July 14, 2025

Participation in the iGBA in HK

• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, Liu Lida, Jia Jingyi
• Purpose: To understand other teams' completion status and research directions
• Method: Attended the exchange meeting
• Outcome: Gained an overview of other teams' progress and refined our own project

July 15, 2025

Attended the IGBA in Shenzhen

• Purpose: Facilitate inter-team communication and understand other teams' progress
• Participants: Wang Xiwu, Wang Yichun, Liu Lida, Xu Wenxuan
• Methods: Booth exchanges, roundtable discussions, corporate presentations
• Outcome: Gained deeper insight into the iGEM project and clarified future development goals

July 21, 2025

Visit to Guangming district Life Sciences Institute

• Purpose: To gain in-depth understanding of industrial production processes for project scale-up reference.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian, Zu Yuhan
• Method: Examining existing commercial large-scale fermentation cultivation technologies
• Results: Gained practical insights into industrial fermentation, providing crucial guidance and reference points for optimizing fermentation conditions in subsequent project phases.

July 25, 2025

Filming promotion video

• Purpose: To promote our project
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi
• Method: Filming with a Nikon Z72 camera and editing using Apple Final Cut Pro X software
• Result: Successfully produced a project promotional video for social media dissemination and competition presentation, enhancing team collaboration and multimedia communication skills.

July 26, 2025

Interviews doctors about astaxanthin products

• Purpose: learn more about current astaxanthin products to write wiki
• Participants: Qin Tian, Xu Wenxuani
• Method: ask four main reasons,which is the relationship of astaxanthin and aging, the antioxident products existed in the market currently, how to maximize the utilization of astaxanthin and how to guarantee safety. Compare the answers with the knowledge we have searched online.
• Outcome: we did not gain too much useful information that we did not know, but this interview verify our previous investigation is correct.

July 28, 2025

Preparing questions for the interview with Beisheng Biotechnology

• Purpose: To prepare clear questions seeking answers
• Participants: Xu Wenxuan, Wang Guanquan
• Method: Collaboratively drafted interview questions, revised and refined after professor's feedback.
• Result: Produced a logically structured, focused interview outline, fully preparing for the subsequent formal interview.

August 2, 2025

Interview with Beisheng Biotechnology

• Purpose: To gain in-depth understanding of the industrialization pathways and technical challenges faced by synthetic biology enterprises.
• Participants: Xu Wenxuan, Wang Xiwu
• Method: Conducted a face-to-face interview with Dr. Chen, focusing on the industrial chain development from R&D to production for his company's products and the challenges encountered.
• Results: Obtained firsthand insights into synthetic biology industrialization. This valuable information provides crucial practical references and optimization strategies for our project direction. One of the most important suggestions that they gave is about business plan。

August 6–8, 2025

Attended CCIC in Beijing

• Purpose: To learn about other teams
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Xu Wenxuan, Wang Xiwu, Wang Yichun, Jia Jingyi
• Method: Attending presentations, booth discussions, introducing our project, exchanging contact information
• Outcome: Gained deeper insight into other teams' progress

August 15, 2025

Interview with Zhongding Testing company

• Purpose: To understand cosmetic regulations and market access for products like astaxanthin.

• Participants: Xu Wenxuan, Wang Xiwu

• Method： We met with CTT, a third-party testing agency, to learn about the legal and safety requirements for cosmetic product launches. This included discussions on compliance, safety assessments, and promotional guidelines.

• Outcome: We gained crucial insights into the NMPA's(National Medical Products Administration) regulatory framework, including restrictions on ingredient claims and new safety evaluation protocols. This guided our reflection on ensuring our project's long-term compliance and product safety.

Art Designing

March 16, 2025

School Emblem Design

• Participants: Wang Xiwu

April 20, 2025

Team Emblem Design

• Participants: Wang Xiwu

May 10, 2025

Team Flag, Uniform, and Materials Design

• Participants: Wang Xiwu, Wang Yichun

May 12, 2025

Poster and Material Production

• Participants: Wang Xiwu, Wang Yichun

August 6, 2025

New Poster Design and Production

• Participants: Wang Xiwu, Wang Yichun

August–September

Wiki Design

• Participants: Wang Xiwu, Wang Yichun

Wiki Development

July 16–18, 2025

Study of 2024 iGEM Team Projects

• Purpose: Compare with our project to identify improvement strategies
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi
• Method: Reviewing award-winning teams' wikis
• Results: By analyzing the design concepts and presentation Methods of outstanding projects, the team gained valuable inspiration and concrete improvement ideas, providing a clear direction for project optimization

July 24, 2025

Drafting the wet lab report

• Purpose: Complete the wiki webpage content framework and deepen understanding of the entire experiment through post-mortem analysis.
• Participants: Wang Guanquan, XU ALAN TSZ HONG
• Method: Conducted a detailed retrospective of past experimental procedures and Outcome images to systematize the overall experimental workflow.
• Results: Successfully established a clear experimental framework, completed the report draft, and enhanced understanding of the project's overall logic.

July 28, 2025

Drafting the Second Version of the Wet Lab Report

• Purpose: To complete the detailed content for the experiment section of the wiki.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Jia Jingyi, Liu Lida, Qin Tian
• Method: Clearly defined responsibilities, with each member assigned specific sections to collaboratively refine experimental procedures and Methodologies.
• Results: Efficiently integrated all experimental data and procedures, completed the second draft of the report, laying a solid foundation for final revisions.

August 15, 2025

Held an online progress coordination meeting

• Purpose: Synchronize Wiki content completion progress, clarify final deadlines and division of labor.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Qin Tian, Wang Yichun, Jia Jingyi
• Method: Each participant reported on the completion status of their respective sections. The team collectively determined the final delivery dates (DDL) for outstanding tasks and assigned responsibilities for collecting PPT materials.
• Outcome: Final deadlines and responsible parties for all tasks were confirmed, effectively enhancing overall team collaboration efficiency and laying the groundwork for timely project completion.

August 20, 2025 – October 5, 2025

Ongoing development and refinement of Wiki content

• Purpose: Focus efforts on completing assigned Wiki sections to ensure content quality and project progress.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Qin Tian, Xu Wenxuan, Wang Yichun, Jia Jingyi, Liu Lida
• Method: During this period, team members intensively developed their assigned page content while holding regular meetings to synchronize progress and collaboratively resolve challenges.
• Outcome: Through high-frequency collaboration and problem-solving, the overall completion of the Wiki was effectively advanced, ensuring both the progress and quality of project documentation.

August 24, 2025

Final Draft of Wet Lab Report

• Purpose: Finalize the experimental content on the wiki.
• Participants: Wang Guanquan, XU ALAN TSZ HONG, Jia Jingyi, Liu Lida, Qin Tian
• Method: Addressed errors and ambiguities based on professor feedback, optimizing logical flow and language.
• Results: Successfully completed an accurate and clearly expressed final report, ready for upload to the team Wiki.

Products

March 30, 2025

Discussed product design

• Purpose: Brainstorm product prototypes
• Participants: Wang Xiwu, Wang Guanquan, XU ALAN TSZ HONG, Qin Tian, Xu Wenxuan, Wang Yichun, Jia Jingyi, Liu Lida
• Method: Team discussion on product design concepts and communication with the supervising instructor
• Outcome: Finalized product as astaxanthin mask

April 21, 2025

Research on past iGEM projects and cosmetic regulations

• Purpose: To learn about product safety considerations and optimize product design.
• Participants: Wang Xiwu, Qin Tian, Xu Wenxuan, Wang Yichun
• Method: Systematic review of product-related pages in past iGEM team wikis and cosmetic production regulations.
• Results: Clarified compliance requirements and safety considerations in product development, providing crucial reference for subsequent design.

May 10, 2025

Color System Conceptualization and Product Ideation

• Purpose: To define our product's identity, inspired by the recent interview revealing a potential consumer base among traditional performers. We aimed to conceptualize the target audience, packaging style, and marketing approach for our astaxanthin-based cosmetic pigment.
• Participants: Wang Xiwu, Qin tian, Xu wenxuan
• Method: Held a dedicated brainstorming session. We discussed how to position the product, drawing from the cultural significance of the color and the unique biotech origin story. Ideas for packaging that blends traditional aesthetics with a modern, scientific feel were sketched and debated.
• Outcome: Established a clear product vision targeting both cultural practitioners and biotech-conscious consumers. Initial packaging concepts featuring minimalist design with accents of our signature "Algal Vermillion" color were drafted. Marketing strategies will emphasize our product's narrative of merging heritage with innovation.

June 13, 2025

Consultation with Advisor

• Purpose: To understand product manufacturing requirements and integrate them with our project
• Participants: Wang Xiwu, Qin Tian, Xu Wenxuan, Wang Yichun
• Method: Listened to explanations, raised questions as a team, and discussed feasibility. Ask whether further research is necessary.
• Outcome: Further refined product design, addressed existing shortcomings, and enhanced project feasibility. Make a appointment to a doctor specialized in astaxanthin one month later.

July 28, 2025

Drafting the initial product design document

• Purpose: Integrate prior discussions and research findings into a comprehensive product design document.
• Participants: Qin Tian
• Method: Draft and revise based on team discussions, instructor feedback, and reference materials
• Result: Completed the initial draft of the product design, providing a comprehensive content foundation for the Wiki module.

August 14, 2025

Gentle Extraction of Astaxanthin

• Purpose: To integrate astaxanthin into the face mask in a safe, mild, and cosmetic matrix-compatible manner, we explored and adopted a food-grade olive oil-based extraction Method.

• Participants: Wang Guanquan, Qin Tian

• Method:

  1. Centrifuge to collect the E. coli cells containing astaxanthin and discard the supernatant.

  2. Freeze cell pellet in liquid nitrogen.

  3. Then, the sample was dried using a vacuum freeze-drying machine. After 48 hours, a powder of bacteria containing astaxanthin was obtained.

  4. Crushed the dried cell pellet into fine powder.

  5. Exactly weigh 10 mg powder into a 2.0 mL centrifuge tube.

  6. Add 0.3mg glass beads (0.5 mm in diameter) and 1 mL of olive oil into the tube.

  7. Use a low-temperature grinder (setting parameters including temperature at 4°C, frequency at 60Hz, cycle at 15-second bursts with 5-second pauses) to break down the cells for 10 minutes.

  8. Centrifuge at 12,000 rpm for 2 minutes at 4°C.

  9. Suck the supernatant with a pipette to a 1.5 mL brown centrifuge tube.

• Result: 'Astaxanthin-Enriched Oil' was successfully prepared. This oil formulation can be directly used as the core active ingredient in face mask solutions, and its mild nature makes it highly suitable for skincare products.

August 15, 2025

Prepare essence and Produce face mask

• Purpose: produce our first masks
• Participants: Qin Tian, Wang Xiwu, Xu Wenxuan
• Method: Prepared essence by mixing the Astaxanthin-Enriched Oil into a base solution. Cut non-woven fabric into facial mask shapes with openings. Placed the fabric into aluminum-plastic bags, injected 25 mL of essence to fully soak it, and heat-sealed the bags after air removal. Finally, packaged the prototypes for stability testing.
• Result: produced 10 pieces of astaxanthin face mask prototypes. The masks exhibited a stable light red colour. 

August 23, 2025

Drafting product security content for the Wiki page

• Purpose: Complete the product safety section of the Wiki.
• Participants: Qin Tian
• Method: Developed content based on product design and team deliverables
• Result: Successfully completed the product safety chapter, ensuring the completeness and compliance of the Wiki content.

The other

March 6, 2025

Team Formation

• Purpose: Establish the core project team and launch the iGEM project.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian, Zu Yuhan
• Method: Team members and preliminary roles were determined through recruitment and consultation.
• Result: Successfully formed a 9-member team, laying the foundation for project implementation.

March 27, 2025

Define Basic Responsibilities for Members

• Purpose: Clarify individual responsibilities to ensure efficient project advancement.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi, Qin Tian, Zu Yuhan
• Method: Preliminary task allocation through self-nomination and team consultation.
• Result: Established general division of labor among members and formed a preliminary collaborative framework.

May 27, 2025

Zu Yuhan withdrew

• Purpose: To adjust team structure in response to evolving project requirements.
• Participants: Zu Yuhan
• Method: Team communication and consultation to redistribute original responsibilities.
• Result: Team members gained deeper understanding of project-related responsibilities and completed the handover smoothly.

July 12, 2025

Team Meeting

• Purpose: Develop a detailed plan for activities over the next month.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, Liu Lida, Jia Jingyi
• Method: Discuss and determine the next steps for mutation research, assigning specific tasks to each member.
• Results: Shared the latest findings from the wet lab experiments and clarified Objectives and responsibilities for the next phase.

July 19, 2025

Study past award-winning projects and prepare for defense

• Purpose: Leverage insights from past award-winning teams to enhance project presentation quality.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi
• Method: Thoroughly research past winning teams' Wiki entries and other materials, prepare PowerPoint presentations, conduct mock defense sessions under Professor Huang's guidance, and refine content based on feedback.
• Results: Identified issues such as excessive presentation time and unfamiliarity with project details. Plan to conduct another mock defense on July 24.

July 23, 2025

Online Mock Defense

• Purpose: To evaluate preparation status and enhance team presentation skills.
• Participants: Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Xu Wenxuan, Jia Jingyi
• Method: Simulated formal defence procedures via online meeting, with team members taking turns presenting and providing mutual feedback.
• Outcome: Gained deeper familiarity with defence procedures, identified and improved deficiencies in presentation.

July 24, 2025

Mock On-Site Defence

• purpose: Deepen understanding of outstanding work characteristics and optimize project presentation content.
• Participants: Xu Wenxuan, Wang Xiwu, Wang Yichun, Wang Guanquan, XU ALAN TSZ HONG, Liu Lida, Jia Jingyi
• Method: Simulated formal defence scenario with timed presentations and Q&A, followed by collective performance analysis.
• Outcome: Enhanced time management and content refinement skills for presentations, preparing the team for the official showcase.