Material

• DH5α (YB-biotech, Taiwan): for plasmid amplification.
• BL21 (YB-biotech, Taiwan): for protein expression of pST39 system
• DStbl3 (YB-biotech, Taiwan): Derived from HB101. It is used for cloning into and storing lentiviral and retroviral vectors for cloning or repeated sequences with the potential to recombine.

• pST39: The pST39 vector is a polycistronic plasmid, and it is used to replicate sequences that form the enzyme and coenzymes producing 5-HTP as well as their motifs. In addition, the vector was also inserted in the PCA promoter and RCR sequence to enhance the production of 5-HTP.

• BioKit Biotechnology Incorporation 1 Kb DNA Ladder
• BioKit Biotechnology Incorporation 100bp DNA ladder
• SMOBIO [PM2600] ExcelBand™ 3-color High Range Protein Marker (9-245 kDa)

• 5-HTP(5-Hydroxytryptophan)ELISA Kit Cat:ELK8124

Primer

DNA gblocks

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DNA gblocks
Zif268	Zinc-finger protein
m-hTPH1	protein that can produce 5-HTP
Zif268-rlinker-m-TPH1	includes Mut-hTPH1 and Zif268 binding domain, with 36 bp linker
PBSII	Zinc-finger protein
hPCBD1	protein acting as an enzyme in the BH4 regeneration system.
PBSII-rlinker-hPCBD1	includes PBSII binding domain and hPCBD1, with 36 bp linker
ZFa	Zinc-finger protein
hQDPR	protein acting as an enzyme in the BH4 regeneration system.
ZFa-hQDPR	includes ZFa binding domain and hQDPR
Codon optimized PcaUAM	protein that can bind on the PCA promoter
pLacIQ-PcaUAM-T7Hyb1-pPCA3B5B-RiboJ-mGL	includes pLacIQ, PcaUAM, T7Hyb1, pPCA3B5B, RiboJ, mGL
pPCA_3B5B	PCA promoter
RiboJ insulator	functional RNA that includes a self-cleaving ribozyme and a hairpin structure to expose the ribosome binding site
pLacIQ	constitutive promoter
RepA	a replication initiator protein
pLacIQ-PcaUAM-T7Hyb1-pPCA3B5B-RiboJ-RepA	includes pLacIQ, PcaUAM, T7Hyb1, pPCA3B5B, RiboJ, RepA.
RCORI-105	the start point of RepA mediated circular ssDNA synthesis in RCR
RCORI-65	the stop point of RepA mediated circular ssDNA synthesis in RCR
mGL	green fluorescent protein
DNA-scaffold-L	part of DNA scaffold
DNA-scaffold-R	part of DNA scaffold

High-amylose starch film:

• High-amylose starch (Premium Warabi Mochi Powder from Japan, purchased from Senguo Foods)
• Glycerol (Origin: Malaysia, purchased from: I CHAN FOOD AND CHEMISTRY COMPANY LTD.)
• Sorbitol (Origin: Thailand, purchased from: I CHAN FOOD AND CHEMISTRY COMPANY LTD.)

Sodium Caseinate Film:

• Sodium caseinate (purchased from: I CHAN FOOD AND CHEMISTRY COMPANY LTD.)
• Glycerol (Origin: Malaysia, purchased from: I CHAN FOOD AND CHEMISTRY COMPANY LTD.)

Reference

1. Diana g calvopina-chavez, Mikaela a gardner, & Joel s griffitts. (2022). Engineering Efficient Termination of Bacteriophage T7 RNA Polymerase Transcription. https://doi.org/10.1093/g3journal/jkac070
2. Sara gonçalves, Daniela nunes-costa, Sandra morais cardoso, Nuno empadinhas, & John david marugg. (2022). Enzyme Promiscuity in Serotonin Biosynthesis, From Bacteria to Plants and Humans. https://doi.org/10.3389/fmicb.2022.873555
3. Jeffrey mckinney, Per m knappskog , Jacinto pereira, Trude ekern , Karen toska, Baukje b kuitert , David levine, Angela m gronenborn, Aurora martinez, & Jan haavik . (2004). Expression and Purification of Human Tryptophan Hydroxylase from Escherichia Coli and Pichia Pastoris. https://doi.org/https://doi.org/10.1016/j.pep.2003.09.014
4. Yuheng lin, Xinxiao sun, Qipeng yuan, & Yajun yan. (2014). Engineering Bacterial Phenylalanine 4-Hydroxylase for Microbial Synthesis of Human Neurotransmitter Precursor 5-Hydroxytryptophan. https://doi.org/10.1021/sb5002505
5. Haijiao wang, Wenqian liu, Feng shi, Lei huang , Jiazhang lian, Liang qu, Jin cai, & Zhinan xu. (2018). Metabolic Pathway Engineering for High-Level Production of 5-Hydroxytryptophan in Escherichia Coli. https://doi.org/10.1016/j.ymben.2018.06.007
6. Meyer, A.J., Segall-Shapiro, T.H., Glassey, E. et al. (2019) Escherichia coli “Marionette” strains with 12 highly optimized small-molecule sensors. . https://doi.org/10.1038/s41589-018-0168-3
7. Jia, Y., et al. (2021, April 8). DNA-Catalyzed Efficient Production of Single-Stranded DNA Nanostructures. ScienceDirect. https://doi.org/10.1016/j.chempr.2020.12.001
8. Conrado, R. J., et al.(2012). DNA-guided assembly of biosynthetic pathways promotes improved catalytic efficiency. Nucleic acids research, 40(4), 1879-1889. https://doi.org/10.1093/nar/gkr888
9. Lin, huei-chih. (2018). Development of Active Edible Sodium Caseinate Packaging Films by Genipin Cross-Linking and Gallic Acid Incorporation.
10. Khaoula khwaldia, Sylvie desobry-banon, Cristina perez-perez, & Stéphane desobry. (2004). Properties of Sodium Caseinate Film-Forming Dispersions and Films. https://doi.org/http://dx.doi.org/10.3168/jds.S0022-0302(04)70018-1
11. Yujia qiu, Yan zhou, Yancheng chang, Xinyue laing, Hui zhang, Xiaorui lin, Ke qing, Xiaojie zhou, & Ziqiang lou. (2022). The Effects of Ventilation, Humidity, and Temperature on Bacterial Growth and Bacterial Genera Distribution. https://doi.org/http://dx.doi.org/10.3390/ijerph192215345
12. Haoxiang wu, & Jonathan wong. (2022). Temperature versus Relative Humidity: Which Is More Important for Indoor Mold Prevention? https://doi.org/http://dx.doi.org/10.3390/jof8070696
13. Xiaoming luo, Haixing song, Jing yang, Bin han, Ye feng, Yanbing leng, & Zhaoqiong chen. (2020). Encapsulation of Escherichia Coli Strain Nissle 1917 in a Chitosan―alginate Matrix by Combining Layer-by-Layer Assembly with CaCl2 Cross-Linking for an Effective Treatment of Inflammatory Bowel Diseases.