癌症為第一的致死原因，五胺基酮戊酸（5-ALA）近年來因其為可治療癌症的光動力治療法的前驅藥物而備受關注。儘管目前已經有許多研究利用微生物產生5-ALA，卻很少有研究專注在密碼子與分子伴侶蛋白對五胺基酮戊酸合成酶（ALAS, EC 2.3.1.37）表達的影響。
在這項研究中，我們分別使用了雙質體、單質體與整合基因組的表達系統於大腸桿菌中生產5-ALA。在雙質體系統中，透過共表達分子伴侶提升可溶ALAS的表達水平，並優化饋料策略將5-ALA的產量提升至5.6 g/L。於單質體系統中，我們利用密碼子混排的技巧證明來自莢膜紅球菌的ALAS表達會隨著密碼子使用頻率的不同而有顯著的差異。藉由分子伴侶GroELS和ALAS共表達及將基因整合至基因組，進一步地提高了細胞耐受性，並且將5-ALA產量提升至5.9 g/L。此外，藉由分析甘胺酸、琥珀酸及磷酸吡哆醛於此系統的重要性，及利用Timer蛋白分析細胞生長狀態找到最佳的饋料時間與條件。再者，由於發現此5-ALA生產系統潛藏著鐵離子與ATP不足的問題，透過添加鐵離子及優化添加條件提升產量與細胞生長，相較於未添加時提升了140 %的生物量與40 %的產量。同時，本研究利用葡萄糖－甘油的混合碳源以延長細胞的生長，1並利用1 L饋料式批次發酵達到15.6 g/L 5-ALA產量。
這項研究全面探討了密碼子對轉譯速度及可溶蛋白表達的影響；利用共表達伴侶蛋白及整合系統提升菌株強度及生長；探討培養基對細胞生長的影響及優化培養基條件，提供了建立微生物細胞工廠的標準程序並得到目前於大腸桿菌生產中的最高產量。

5‐aminolevulinic acid (5-ALA) is a high-value chemical and has attracted much attention in recent years because it is a precursor drug for photodynamic therapy that can treat cancer. Although there have been several studies using microorganisms to produce 5-ALA, only a few researches focused on the effect of codons and chaperones on the expression of 5‐aminolevulinic acid synthase (ALAS, EC 2.3.1.37).
In this study, we used dual-plasimd system, single-plasmid system, and genome-based expression system to produce 5-ALA. In the dual-plasmid system, the expression level of soluble ALAS was increased by co-expression of chaperone GroELS, and the feeding strategy was optimized to enhance the titer to 5.6 g/L. In the single-plasmid system, we used the technique of codon shuffling to prove the importance of codons on the expression of ALAS from Rhodobactor capsulatus. Then, the co-expression of the chaperone GroELS and the genome-based expression system further improved the 5-ALA production to 5.9 g/L. Furthermore, the addition of ferric citrate with optimal conditions increased 140 % biomass and 40 % 5-ALA production. At the same time, the 5-ALA titer was enhanced to 15.6 g/L in 1 L fed-batch fermentation using glucose-glycerol mixed carbon source. In this study, we provided a standard process to establish an economic and highly efficient biofinery for high-value chemical production.

Ambrus, J. L., Ambrus, C. M., Mink, I. B., & Pickren, J. W. Causes of death in cancer patients. Journal of Medicine, 6(1), 61-64. (1975).
Agirrezabala, X., & Frank, J. Elongation in translation as a dynamic interaction among the ribosome, tRNA, and elongation factors EF-G and EF-Tu. Quarterly Reviews of Biophysics, 42(3), 159. (2009).
Akram, N. A., & Ashraf, M. Regulation in plant stress tolerance by a potential plant growth regulator, 5-aminolevulinic acid. Journal of Plant Growth Regulation, 32(3), 663-679. (2013).
Ali, B., Wang, B., Ali, S., Ghani, M. A., Hayat, M. T., Yang, C., Xu, L. & Zhou, W. J. 5-Aminolevulinic acid ameliorates the growth, photosynthetic gas exchange capacity, and ultrastructural changes under cadmium stress in Brassica napus L. Journal of Plant Growth Regulation, 32(3), 604-614. (2013).
Beale, S. I. The biosynthesis of δ-aminolevulinic acid in Chlorella. Plant Physiology, 45, 504-506. (1970).
Briggs, M. R., Kadonaga, J. T., Bell, S. P., & Tjian, R. Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science, 234(4772), 47-52. (1986).
Boni, I. V., lsaeva, D. M., Musychenko, M. L., & Tzareva, N. V. Ribosome-messenger recognition: mRNA target sites for ribosomal protein S1. Nucleic Acids Research, 19(1), 155-162. (1991).
Bozell, J. J., Moens, L., Elliott, D. C., Wang, Y., Neuenscwander, G. G., Fitzpatrick, S. W., Bilski, R. J. & Jarnefeld, J. L. Production of levulinic acid and use as a platform chemical for derived products. Resources, Conservation and Recycling, 28(3-4), 227-239. (2000).
Bartošová, J., & Hrkal, Z. Accumulation of protoporphyrin-IX (PpIX) in leukemic cell lines following induction by 5-aminolevulinic acid (ALA). Comparative Biochemistry and Physiology, 126(3), 245-252. (2000).
Bunke, A., Zerbe, O., Schmid, H., Burmeister, G., Merkle, H. P., & Gander, B. Degradation mechanism and stability of 5‐aminolevulinic acid. Journal of Pharmaceutical Sciences, 89(10), 1335-1341. (2000).
Blanchard, S. C., Kim, H. D., Gonzalez, R. L., Puglisi, J. D., & Chu, S. tRNA dynamics on the ribosome during translation. Proceedings of the National Academy of Sciences, 101(35), 12893-12898. (2004).
Boström, M., Markland, K., Sandén, A. M., Hedhammar, M., Hober, S., & Larsson, G. Effect of substrate feed rate on recombinant protein secretion, degradation and inclusion body formation in Escherichia coli. Applied Microbiology and Biotechnology, 68(1), 82-90. (2005).
Bren, A., Park, J. O., Towbin, B. D., Dekel, E., Rabinowitz, J. D., & Alon, U. Glucose becomes one of the worst carbon sources for E. coli on poor nitrogen sources due to suboptimal levels of cAMP. Scientific Reports, 6(1), 1-10. (2016).
Brown, B. L., Kardon, J. R., Sauer, R. T., & Baker, T. A. Structure of the mitochondrial aminolevulinic acid synthase, a key heme biosynthetic enzyme. Structure, 26(4), 580-589. (2018).
Bolognesi, B., & Lehner, B. Protein overexpression: reaching the limit. Elife, 7, e39804. (2018).
Boyaci, H., Chen, J., Jansen, R., Darst, S. A., & Campbell, E. A. Structures of an RNA polymerase promoter melting intermediate elucidate DNA unwinding. Nature, 565(7739), 382-385. (2019).
Cooper, S., & Helmstetter, C. E. Chromosome replication and the division cycle of Escherichia coli Br. Journal of Molecular Biology, 31(3), 519-540. (1968).
Challice, J. M., & Segall, J. Transcription of the 5 S rRNA gene of Saccharomyces cerevisiae requires a promoter element at+ 1 and a 14-base pair internal control region. Journal of Biological Chemistry, 264(33), 20060-20067. (1989).
Chen, J., Keltner, L., Christophersen, J., Zheng, F., Krouse, M., Singhal, A., & Wang, S. S. New technology for deep light distribution in tissue for phototherapy. The Cancer Journal, 8(2), 154-163. (2002).
Carbone, A., Zinovyev, A., & Képes, F. Codon adaptation index as a measure of dominating codon bias. Bioinformatics, 19(16), 2005-2015. (2003).
Collaud, S., Juzeniene, A., Moan, J., & Lange, N. On the selectivity of 5-aminolevulinic acid-induced protoporphyrin IX formation. Current Medicinal Chemistry-Anti-Cancer Agents, 4(3), 301-316. (2004).
Cartharius, K., Frech, K., Grote, K., Klocke, B., Haltmeier, M., Klingenhoff, A., Frisch, M., Bayerlein, M. & Werner, T. MatInspector and beyond: promoter analysis based on transcription factor binding sites. Bioinformatics, 21(13), 2933-2942. (2005).
Casas, A., & Batlle, A. Aminolevulinic acid derivatives and liposome delivery as strategies for improving 5-aminolevulinic acid-mediated photodynamic therapy. Current Medicinal Chemistry, 13(10), 1157-1168. (2006).
Choi, H. P., Lee, Y. M., Yun, C. W., & Sung, H. C. Extracellular 5-aminolevulinic acid production by Escherichia coli containing the Rhodopseudomonas palustris KUGB306 hemA gene. Journal of Microbiology and Biotechnology, 18(6), 1136-1140. (2008).
Cannarozzi, G., Schraudolph, N. N., Faty, M., von Rohr, P., Friberg, M. T., Roth, A. C., Gonnet, P., Gonnet, G. & Barral, Y. A role for codon order in translation dynamics. Cell, 141(2), 355-367. (2010).
Claudi, B., Spröte, P., Chirkova, A., Personnic, N., Zankl, J., Schürmann, N., Schmidt, A. & Bumann, D. Phenotypic variation of Salmonella in host tissues delays eradication by antimicrobial chemotherapy. Cell, 158(4), 722-733. (2014).
Chen, H., Shiroguchi, K., Ge, H., & Xie, X. S. Genome‐wide study of mRNA degradation and transcript elongation in Escherichia coli. Molecular Systems Biology, 11(1), 781. (2015).
Cui, Z., Jiang, Z., Zhang, J., Zheng, H., Jiang, X., Gong, K., Liang, Q., Wang, Q., Qi, Q. Stable and efficient biosynthesis of 5-aminolevulinic acid using plasmid-free Escherichia coli. Journal of Agricultural and Food Chemistry, 67, 1478-1483. (2019).
Chen, J., Wang, Y., Guo, X., Rao, D., Zhou, W., Zheng, P., & Ma, Y. Efficient bioproduction of 5-aminolevulinic acid, a promising biostimulant and nutrient, from renewable bioresources by engineered Corynebacterium glutamicum. Biotechnology for Biofuels, 13(1), 1-13. (2020).
Chiang, C. J., Hu, M. C., & Chao, Y. P. A. Strategy to improve production of recombinant proteins in Escherichia coli based on a glucose–glycerol mixture and glutamate. Journal of Agricultural and Food Chemistry, 68(33), 8883-8889. (2020).
de Marco, A., Deuerling, E., Mogk, A., Tomoyasu, T., & Bukau, B. Chaperone-based procedure to increase yields of soluble recombinant proteins produced in E. coli. BMC Biotechnology, 7(1), 1-9. (2007).
Da Silva, G. P., Mack, M., & Contiero, J. Glycerol: a promising and abundant carbon source for industrial microbiology. Biotechnology Advances, 27(1), 30-39. (2009).
Dvorak, P., Chrast, L., Nikel, P. I., Fedr, R., Soucek, K., Sedlackova, M., Chaloupkova, R., de Lorenzo, V., Prokop, Z. & Damborsky, J. Exacerbation of substrate toxicity by IPTG in Escherichia coli BL21 (DE3) carrying a synthetic metabolic pathway. Microbial Cell Factories, 14(1), 1-15. (2015).
El-Mansi, E. M. T., & Holms, W. H. Control of carbon flux to acetate excretion during growth of Escherichia coli in batch and continuous cultures. Microbiology, 135(11), 2875-2883. (1989).
Elfsson, B., Wallin, I., Eksborg, S., Rudaeus, K., Ros, A. M., & Ehrsson, H. Stability of 5-aminolevulinic acid in aqueous solution. European Journal of Pharmaceutical Sciences, 7(2), 87-91. (1999).
Effendi, S. S. W., Tan, S. I., Ting, W. W., & Ng, I. S. Enhanced recombinant Sulfurihydrogenibium yellowstonense carbonic anhydrase activity and thermostability by chaperone GroELS for carbon dioxide biomineralization. Chemosphere, 128461. (2020).
Fritsch, C., Verwohlt, B., Bolsen, K., Ruzicka, T., & Goerz, G. Influence of topical photodynamic therapy with 5-aminolevulinic acid on porphyrin metabolism. Archives of Dermatological Research, 288(9), 517-521. (1996).
Fink, A. L. Protein aggregation: folding aggregates, inclusion bodies and amyloid. Folding and Design, 3(1), R9-R23. (1998).
Fotinos, N., Campo, M. A., Popowycz, F., Gurny, R., & Lange, N. 5‐Aminolevulinic acid derivatives in photomedicine: Characteristics, application and perspectives. Photochemistry and Photobiology, 82(4), 994-1015. (2006).
Fu, W., Lin, J., & Cen, P. Enhancement of 5-aminolevulinate production with recombinant Escherichia coli using batch and fed-batch culture system. Bioresource Technology, 99(11), 4864-4870. (2008).
Fu, W., Lin, J., & Cen, P. Expression of a hemA gene from Agrobacterium radiobacter in a rare codon optimizing Escherichia coli for improving 5-aminolevulinate production. Applied Biochemistry and Biotechnology, 160(2), 456-466. (2010).
Fischer, N., Konevega, A. L., Wintermeyer, W., Rodnina, M. V., & Stark, H. Ribosome dynamics and tRNA movement by time-resolved electron cryomicroscopy. Nature, 466(7304), 329-333. (2010).
Green, R., & Noller, H. F. Ribosomes and translation. Annual Review of Biochemistry, 66(1), 679-716. (1997).
Gadmar, Ø. B., Moan, J., Scheie, E., Ma, L. W., & Peng, Q. The stability of 5-aminolevulinic acid in solution. Journal of Photochemistry and Photobiology B: Biology, 67(3), 187-193. (2002).
Gupta, P., Aggarwal, N., Batra, P., Mishra, S., & Chaudhuri, T. K. Co-expression of chaperonin GroEL/GroES enhances in vivo folding of yeast mitochondrial aconitase and alters the growth characteristics of Escherichia coli. The International Journal of Biochemistry and Cell Biology, 38(11), 1975-1985. (2006).
Gatti‐Lafranconi, P., Natalello, A., Ami, D., Doglia, S. M., & Lotti, M. Concepts and tools to exploit the potential of bacterial inclusion bodies in protein science and biotechnology. The FEBS Journal, 278(14), 2408-2418. (2011).
Hoffmann, B., Messer, W., & Schwarz, U. Regulation of polar cap formation in the life cycle of Escherichia coli. Journal of Supramolecular Structure, 1(1), 29-37. (1972).
Horn, U., Strittmatter, W., Krebber, A., Knüpfer, U., Kujau, M., Wenderoth, R., Müller, K., Matzku, S., Plückthun, A. & Riesenberg, D. High volumetric yields of functional dimeric miniantibodies in Escherichia coli, using an optimized expression vector and high-cell-density fermentation under non-limited growth conditions. Applied Microbiology and Biotechnology, 46(5), 524-532. (1996).
Hotta, Y., Tanaka, T., Takaoka, H., Takeuchi, Y., & Konnai, M. New physiological effects of 5-aminolevulinic acid in plants: the increase of photosynthesis, chlorophyll content, and plant growth. Bioscience, Biotechnology, and Biochemistry, 61(12), 2025-2028. (1997).
Hirata, H., Yamamura, I., Yasuda, K., Kobayashi, A., Tada, N., Suzuki, M., Hirayoshi, K., Hosokawa, N. & Nagata, K. Separate cis-acting DNA elements control cell type-and tissue-specific expression of collagen binding molecular chaperone HSP47. Journal of Biological Chemistry, 274(50), 35703-35710. (1999).
Hongcharu, W., Taylor, C. R., Aghassi, D., Suthamjariya, K., Anderson, R. R., & Chang, Y. Topical ALA-photodynamic therapy for the treatment of acne vulgaris. Journal of Investigative Dermatology, 115(2), 183-192. (2000).
Hellen, C. U., & Sarnow, P. Internal ribosome entry sites in eukaryotic mRNA molecules. Genes and Development, 15(13), 1593-1612. (2001).
Heinemann, I. U., Jahn, M., & Jahn, D. The biochemistry of heme biosynthesis. Archives of Biochemistry and Biophysics, 474(2), 238-251. (2008).
Hayer-Hartl, M., Bracher, A., & Hartl, F. U. The GroEL–GroES chaperonin machine: a nano-cage for protein folding. Trends in Biochemical Sciences, 41(1), 62-76. (2016).
Ito, K., Uno, M., & Nakamura, Y. A tripeptide ‘anticodon’ deciphers stop codons in messenger RNA. Nature, 403(6770), 680-684. (2000).
Johnston, D. Stages of translation. Bath: Absolute classics. (1996).
Jung, S. K., & McDonald, K. Visual gene developer: a fully programmable bioinformatics software for synthetic gene optimization. Bmc Bioinformatics, 12(1), 1-13. (2011).
Kitani, A., Tsunetsugu, S. I., & Sasaki, K. Fe III-ion-catalysed non-enzymatic transformation of ADP in to ATP. Journal of the Chemical Society, Perkin Transactions 2, (3), 329-331. (1991).
Korz, D. J., Rinas, U., Hellmuth, K., Sanders, E. A., & Deckwer, W. D. Simple fed-batch technique for high cell density cultivation of Escherichia coli. Journal of Biotechnology, 39(1), 59-65. (1995).
Kane, J. F. Effects of rare codon clusters on high-level expression of heterologous proteins in Escherichia coli. Current Opinion in Biotechnology, 6(5), 494-500. (1995).
Keskin, O., Bahar, I., Flatow, D., Covell, D. G., & Jernigan, R. L. Molecular mechanisms of chaperonin GroEL− GroES function. Biochemistry, 41(2), 491-501. (2002).
Kang, Z., Wang, Y., Wang, Q., & Qi, Q. Metabolic engineering to improve 5-aminolevulinic acid production. Bioengineered Bugs, 2(6), 342-345. (2011).
Kang, Z., Wang, Y., Gu, P., Wang, Q., Qi, Q. Engineering Escherichia coli for efficient production of 5-aminolevulinic acid from glucose. Metabolic Engineering, 13, 492-498. (2011).
Kim, Y. E., Hipp, M. S., Bracher, A., Hayer-Hartl, M., & Ulrich Hartl, F. Molecular chaperone functions in protein folding and proteostasis. Annual Review of Biochemistry, 82, 323-355. (2013).
Kang, Z., Ding, W., Gong, X., Liu, Q., Du, G., & Chen, J. Recent advances in production of 5‐aminolevulinic acid using biological strategies. World Journal of Microbiology and Biotechnology, 33, 200. (2017).
Ko, Y. J., You, S. K., Kim, M., Lee, E., Shin, S. K., Park, H. M., Oh, Y., Han, S. O. Enhanced production of 5-aminolevulinic acid via flux redistribution of TCA cycle toward l-glutamate in Corynebacterium glutamicum. Biotechnology and Bioprocess Engineering, 24(6), 915-923. (2019).
Lin, J., Fu, W., & Cen, P. Characterization of 5-aminolevulinate synthase from Agrobacterium radiobacter, screening new inhibitors for 5-aminolevulinate dehydratase from Escherichia coli and their potential use for high 5-aminolevulinate production. Bioresource Technology, 100(7), 2293-2297. (2009).
Li, F., Wang, Y., Gong, K., Wang, Q., Liang, Q., Qi, Q. Constitutive expression of RyhB regulates the heme biosynthesis pathway and increases the 5-aminolevulinic acid accumulation in Escherichia coli. FEMS Microbiology Letters, 350, 209-215. (2014).
Liu, S., Zhang, G., Li, X., & Zhang, J. Microbial production and applications of 5-aminolevulinic acid. Applied Microbiology and Biotechnology, 98(17), 7349-7357. (2014).
Lou, J. W., Zhu, L., Wu, M. B., Yang, L. R., Lin, J. P., & Cen, P. L. High-level soluble expression of the hemA gene from Rhodobacter capsulatus and comparative study of its enzymatic properties. Journal of Zhejiang University SCIENCE B, 15(5), 491-499. (2014).
Liu, S., Zhang, G., Li, J., Li, X., & Zhang, J. Effects of metal ions on biomass and 5-aminolevulinic acid production in Rhodopseudomonas palustris wastewater treatment. Water Science and Technology, 73(2), 382-388. (2016).
Ling, X. U., Islam, F., Zhang, W. F., Ghani, M. A., & Ali, B. 5-aminolevulinic acid alleviates herbicide-induced physiological and ultrastructural changes in Brassica napus. Journal of Integrative Agriculture, 17(3), 579-592. (2018).
Liu, S., Zheng, Z., Tie, J., Kang, J., Zhang, G., & Zhang, J. Impacts of Fe2+ on 5-aminolevulinic acid (ALA) biosynthesis of Rhodobacter sphaeroides in wastewater treatment by regulating nif gene expression. Journal of Environmental Sciences, 70, 11-19. (2018).
Mauzerall, D., & Granick, S. The occurrence and determination of 5-aminolevulinic acid and porphobilinogen in urine. Journal of Biological Chemistry, 219(156), 435-446. (1956).
Martin, J. L. Thioredoxin—a fold for all reasons. Structure, 3(3), 245-250. (1995).
Mirabella, R., Franken, C., van der Krogt, G. N., Bisseling, T., & Geurts, R. Use of the fluorescent timer DsRED-E5 as reporter to monitor dynamics of gene activity in plants. Plant Physiology, 135(4), 1879-1887. (2004).
Man, O., Sussman, J. L., & Pilpel, Y. Examination of the tRNA adaptation index as a predictor of protein expression levels. In Systems Biology and Regulatory Genomics, 107-118. (2005).
Martinez-Hackert, E., & Hendrickson, W. A. Promiscuous substrate recognition in folding and assembly activities of the trigger factor chaperone. Cell, 138(5), 923-934. (2009).
Mascal, M., & Dutta, S. Synthesis of the natural herbicide δ-aminolevulinic acid from cellulose-derived 5-(chloromethyl) furfural. Green Chemistry, 13(1), 40-41. (2011).
Mitarai N & Pedersen S. Control of ribosome traffic by position-dependent choice of synonymous codons. Physical Biology, 10(5), 056011. (2013).
Moon, Y. M., Yang, S. Y., Choi, T. R., Jung, H. R., Song, H. S., Han, Y., Park, H. Y., Bhatia, S. K., Gurav, R., Park, K., Kim, J. S. & Yang, Y. H. Enhanced production of cadaverine by the addition of hexadecyltrimethylammonium bromide to whole cell system with regeneration of pyridoxal-5’-phosphate and ATP. Enzyme and Microbial Technology, 127, 58-64. (2019).
Miscevic, D., Mao, J. Y., Kefale, T., Abedi, D., Moo‐Young, M., & Chou, C. P. Strain engineering for high‐level 5‐aminolevulinic acid production in Escherichia coli. Biotechnology and Bioengineering, 1-13. (2021).
Novoa, E. M., Pavon-Eternod, M., Pan, T., & de Pouplana, L. R. A role for tRNA modifications in genome structure and codon usage. Cell, 149(1), 202-213. (2012).
Orenstein, A., Kostenich, G., Roitman, L., Shechtman, Y., Kopolovic, Y., Ehrenberg, B., & Malik, Z. A comparative study of tissue distribution and photodynamic therapy selectivity of chlorin e6, Photofrin II and ALA-induced protoporphyrin IX in a colon carcinoma model. British Journal of Cancer, 73(8), 937-944. (1996).
Oleinick, N. L., & Evans, H. H. The photobiology of photodynamic therapy: cellular targets and mechanisms. Radiation Research, 150(5), 146-156. (1998).
Oh, M. K., & Liao, J. C. DNA microarray detection of metabolic responses to protein overproduction in Escherichia coli. Metabolic Engineering, 2(3), 201-209. (2000).
Perrett, S., Zahn, R., Stenberg, G., & Fersht, A. R. Importance of electrostatic interactions in the rapid binding of polypeptides to GroEL. Journal of Molecular Biology, 269(5), 892-901. (1997).
Park, S. J., Kim, E. Y., Noh, W., Park, H. M., Oh, Y. H., Lee, S. H., Song, B. K., Jegal, J. & Lee, S. Y. Metabolic engineering of Escherichia coli for the production of 5-aminovalerate and glutarate as C5 platform chemicals. Metabolic Engineering, 16, 42-47. (2013).
Quax, T. E., Claassens, N. J., Söll, D., & van der Oost, J. Codon bias as a means to fine-tune gene expression. Molecular Cell, 59(2), 149-161. (2015).
Rebeiz, C. A., Juvik, J. A., Rebeiz, C. C., Bouton, C. E., & Gut, L. J. Porphyric insecticides: 2. 1, 10-Phenanthroline, a potent porphyric insecticide modulator. Pesticide Biochemistry and Physiology, 36(2), 201-207. (1990).
Rebeiz, C. A., Amindari, S., Reddy, K. N., Nandihalli, U. B., Moubarak, M. B., & Velu, J. A. δ-Aminolevulinic acid based herbicides and tetrapyrrole biosynthesis modulators. Porphyric Pesticides, 4, 48-64. (1994).
Ryter, S. W., & Tyrrell, R. M. The heme synthesis and degradation pathways: role in oxidant sensitivity: heme oxygenase has both pro-and antioxidant properties. Free Radical Biology and Medicine, 28(2), 289-309. (2000).
Ramakrishnan, V. Ribosome structure and the mechanism of translation. Cell, 108(4), 557-572. (2002).
Ramzi, A. B., Hyeon, J. E., Kim, S. W., Park, C., & Han, S. O. 5-aminolevulinic acid production in engineered Corynebacterium glutamicum via C5 biosynthesis pathway. Enzyme and Microbial Technology, 81, 1-7. (2015).
Ren, J., Zhou, L., Wang, C., Lin, C., Li, Z., & Zeng, A. P. An unnatural pathway for efficient 5-aminolevulinic acid biosynthesis with glycine from glyoxylate based on retrobiosynthetic design. ACS Synthetic Biology, 7(12), 2750-2757. (2018).
Shiloach, J., & Bauer, S. High‐yield growth of E. coli at different temperatures in a bench scale fermentor. Biotechnology and Bioengineering, 17(2), 227-239. (1975).
Schröder, H., Langer, T., Hartl, F. U., & Bukau, B. DnaK, DnaJ and GrpE form a cellular chaperone machinery capable of repairing heat‐induced protein damage. The EMBO Journal, 12(11), 4137-4144. (1993).
Sasaki, K., Watanabe, M., & Tanaka, T. Biosynthesis, biotechnological production and applications of 5-aminolevulinic acid. Applied Microbiology and Biotechnology, 58(1), 23-29. (2002).
Szeimies, R. M., & Landthaler, M. Photodynamic therapy and fluorescence diagnosis of skin cancers. Cancers of the Skin, 240-245. (2002).
Szeimies, R. M., Landthaler, M., & Karrer, S. Non-oncologic indications for ALA-PDT. Journal of Dermatological Treatment, 13(sup1), s13-s18. (2002).
Switka-Wieclawska, I., Kecik, T., Kwasny, M., & Graczyk, A. Photodynamic therapy (ALA-PDT) in the treatment of pathological states of the cornea. In Laser Technology VII: Applications of Lasers, 5229, 82-86. (2003).
Shiloach, J., & Fass, R. Growing E. coli to high cell density—a historical perspective on method development. Biotechnology Advances, 23(5), 345-357. (2005).
Susin, M. F., Baldini, R. L., Gueiros-Filho, F., & Gomes, S. L. GroES/GroEL and DnaK/DnaJ have distinct roles in stress responses and during cell cycle progression in Caulobacter crescentus. Journal of Bacteriology, 188(23), 8044-8053. (2006).
Stoletzki, N., & Eyre-Walker, A. Synonymous codon usage in Escherichia coli: selection for translational accuracy. Molecular Biology and Evolution, 24(2), 374-381. (2007).
Su, T., Guo, Q., Zheng, Y., Liang, Q., Wang, Q., Qi, Q. Fine-tuning of hemB using CRISPRi for increasing 5-aminolevulinic acid production in Escherichia coli. Frontiers in Microbiology, 10. (2019).
Shih, I. T. 大腸桿菌生產五胺基酮戊酸之蛋白表達模組及代謝調控. 成功大學化學工程學系學位論文, 1-83. (2020).
Thissen, M. R. T. M., Schroeter, C. A., & Neumann, H. A. M. Photodynamic therapy with delta‐aminolaevulinic acid for nodular basal cell carcinomas using a prior debulking technique. British Journal of Dermatology, 142(2), 338-339. (2000).
Tangprasittipap, A., & Prasertsan, P. 5-aminolevulinic acid from photosynthetic bacteria and its applications. Songklanakarin Journal of Science and Technology, 24(4), 717-725. (2002).
Tsai, T., Ji, H. T., Chiang, P. C., Chou, R. H., Chang, W. S. W., & Chen, C. T. ALA‐PDT results in phenotypic changes and decreased cellular invasion in surviving cancer cells. The Official Journal of the American Society for Laser Medicine and Surgery, 41(4), 305-315. (2009).
Tuller, T., Carmi, A., Vestsigian, K., Navon, S., Dorfan, Y., Zaborske, J., Pan, T., Dahen, O., Furman, I. & Pilpel, Y. An evolutionarily conserved mechanism for controlling the efficiency of protein translation. Cell, 141(2), 344-354. (2010).
Todeschini, A. L., Georges, A., & Veitia, R. A. Transcription factors: specific DNA binding and specific gene regulation. Trends in Genetics, 30(6), 211-219. (2014).
Uehlinger, P., Zellweger, M., Wagnières, G., Juillerat-Jeanneret, L., van den Bergh, H., & Lange, N. 5-Aminolevulinic acid and its derivatives: physical chemical properties and protoporphyrin IX formation in cultured cells. Journal of Photochemistry and Photobiology, 54(1), 72-80. (2000).
Woldringh, C. L. Morphological analysis of nuclear separation and cell division during the life cycle of Escherichia coli. Journal of Bacteriology, 125(1), 248-257. (1976).
Weber, A. L., & Lacey, J. C. Genetic code correlations: amino acids and their anticodon nucleotides. Journal of Molecular Evolution, 11(3), 199-210. (1978).
Wang, X. L., Wang, H. W., Guo, M. X., & Xu, S. Z. Treatment of skin cancer and pre-cancer using topical ALA-PDT—a single hospital experience. Photodiagnosis and Photodynamic Therapy, 5(2), 127-133. (2008).
White, R. J. Gene transcription: mechanisms and control. John Wiley & Sons. (2009).
Wu, P., Zhang, G., Li, J., Lu, H., & Zhao, W. Effects of Fe2+ concentration on biomass accumulation and energy metabolism in photosynthetic bacteria wastewater treatment. Bioresource Technology, 119, 55-59. (2012).
Wu, Y., Liao, W., Dawuda, M. M., Hu, L., & Yu, J. 5-Aminolevulinic acid (ALA) biosynthetic and metabolic pathways and its role in higher plants: a review. Plant Growth Regulation, 87(2), 357-374. (2019).
Wen, Q., Liu, B., Li, F., & Chen, Z. Substrate strategy optimization for polyhydroxyalkanoates producing culture enrichment from crude glycerol. Bioresource Technology, 311, 123516. (2020).
Xu, Z., Horwich, A. L., & Sigler, P. B. The crystal structure of the asymmetric GroEL–GroES–(ADP) 7 chaperonin complex. Nature, 388(6644), 741-750. (1997).
Xie, L., Wang, Z. H., Cheng, X. H., Gao, J. J., Zhang, Z. P., & Wang, L. J. 5-Aminolevulinic acid promotes anthocyanin accumulation in Fuji apples. Plant Growth Regulation, 69(3), 295-303. (2013).
Xue, C., Hsu, K. M., Ting, W. W., Huang, S. F., Lin, H. Y., Li, S. F., Chang, J. S. & Ng, I. S. Efficient biotransformation of L-lysine into cadaverine by strengthening pyridoxal 5’-phosphate-dependent proteins in Escherichia coli with cold shock treatment. Biochemical Engineering Journal, 161, 107659. (2020).
Yasukawa, T., Kanei-Ishii, C., Maekawa, T., Fujimoto, J., Yamamoto, T., & Ishii, S. Increase of solubility of foreign proteins in Escherichia coli by coproduction of the bacterial thioredoxin. Journal of Biological Chemistry, 270(43), 25328-25331. (1995).
Yang, M. L., Kun, Y. I. N., Guo, Y. P., & Zhang, J. Z. A photosensitivity insecticide, 5-aminolevulinic acid, exerts effective toxicity to oxya chinensis (Orthoptera: Acridoidea). Agricultural Sciences in China, 10(7), 1056-1063. (2011).
You, C., Zhang, X. Z., & Zhang, Y. H. P. Simple cloning via direct transformation of PCR product (DNA Multimer) to Escherichia coli and Bacillus subtilis. Applied and Environmental Microbiology, 78(5), 1593-1595. (2012).
Yang, J., Zhu, L., Fu, W., Lin, Y., Lin, J. P., & Cen, P. L. Improved 5-aminolevulinic acid production with recombinant Escherichia coli by a short-term dissolved oxygen shock in fed-batch fermentation. Chinese Journal of Chemical Engineering, 21(11), 1291-1295. (2013).
Yang, X., Li, W., Palasuberniam, P., Myers, K. A., Wang, C., & Chen, B. Effects of silencing heme biosynthesis enzymes on 5‐aminolevulinic acid‐mediated protoporphyrin IX fluorescence and photodynamic therapy. Photochemistry and Photobiology, 91(4), 923-930. (2015).
Yu, X., Jin, H., Liu, W., Wang, Q., & Qi, Q. Engineering Corynebacterium glutamicum to produce 5-aminolevulinic acid from glucose. Microbial Cell Factories, 14(1), 1-10. (2015a).
Yu, C. H., Dang, Y., Zhou, Z., Wu, C., Zhao, F., Sachs, M. S., & Liu, Y. Codon usage influences the local rate of translation elongation to regulate co-translational protein folding. Molecular Cell, 59(5), 744-754. (2015b).
Yang, P., Liu, W., Cheng, X., Wang, J., Wang, Q., & Qi, Q. A new strategy for production of 5-aminolevulinic acid in recombinant Corynebacterium glutamicum with high yield. Applied and Environmental Microbiology, 82(9), 2709-2717. (2016).
Yu, T. H., Yi, Y. C., Shih, I. T., & Ng, I. S. Enhanced 5-aminolevulinic acid production by co-expression of codon-optimized hemA gene with chaperone in genetic engineered Escherichia coli. Applied Biochemistry and Biotechnology, 1-14. (2019).
Ye, H., He, Y., Xie, Y., Sen, B., & Wang, G. Fed-batch fermentation of mixed carbon source significantly enhances the production of docosahexaenoic acid in Thraustochytriidae sp. PKU# Mn16 by differentially regulating fatty acids biosynthetic pathways. Bioresource Technology, 297, 122402. (2020).
Zeng, X., Chen, X. S., Gao, Y., Ren, X. D., Wang, L., & Mao, Z. G. Continuously high reactive oxygen species generation decreased the specific ϵ-poly-l-lysine formation rate in fed-batch fermentation using glucose and glycerol as a mixed carbon source. Process Biochemistry, 50(12), 1993-2003. (2015).
Zhang, J., Weng, H., Ding, W., & Kang, Z. N-terminal engineering of glutamyl-tRNA reductase with positive charge arginine to increase 5-aminolevulinic acid biosynthesis. Bioengineered, 8(4), 424-427. (2017).
Zhao, M., Li, G., & Deng, Y. Engineering Escherichia coli for glutarate production as the C5 platform backbone. Applied and Environmental Microbiology, 84(16). (2018).
Zhu, C., Chen, J., Wang, Y., Wang, L., Guo, X., Chen, N., & Ma, Y. Enhancing 5-aminolevulinic acid tolerance and production by engineering the antioxidant defense system of Escherichia coli. Biotechnology and Bioengineering, 116(8), 2018-2028. (2019).
Zhang, J., Kang, Z., Ding, W., Chen, J., Du, G. Integrated optimization of the in vivo heme biosynthesis pathway and the in vitro iron concentration for 5-aminolevulinate production. Applied Biochemistry and Biotechnology, 178, 1252-1262. (2016).
Zhang, J., Weng, H., Zhou, Z., Du, G., Kang, Z. Engineering of multiple modular pathways for high-yield production of 5-aminolevulinic acid in Escherichia coli. Bioresource Technology, 274, 353-360. (2019).
迟鹏利, 谢崔越, & 林吉茂. (2013). 5-氨基酮戊酸盐酸盐的合成新方法. 有机化学, 3.