Part:BBa_K3612009
Tenebrio molitor antifreeze protein (TmAFP)
TmAFP is the antifreeze protein from the insect Tenebrio molitor, and given that it has high TH activity, our team chose it as one of the AFPs to be expressed in Pseudoalteromonas nigrifaciens. The expression in the marine bacteria of Pseudoalteromonas genus will allow us to produce TmAFP in cold and low-tech environments, like the Andean Peruvian region, so that our antifreeze product based on AFPs could be at farmers disposal.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Contents
Usage
The antifreeze proteins (AFPs) are used as cryoprotectants in various fields, mainly in medicine and food industry (1, 2). They are considered as a safer option compared to the traditional chemical solutions such as DMSO or liquid nitrogen, and constitute an easy method compared to the laborious cryopreservation processes (1, 2).
In recent years, its applications have widely extended even to the agriculture field (1). Recent studies have successfully modified plants of tobacco, tomato and Arabidopsis thaliana to express AFPs resulting in plants resistant to low temperatures (3-5). Our team also wants to use AFPs to protect crops from cold injury by developing an antifreeze product based on antifreeze proteins.
Biology
The antifreeze proteins are a type of ice binding proteins (IBPs), which adhere to the surface of an ice crystal and follow an adsorption-inhibition mechanism, preventing ice crystal growth. Its function is allowed by two main properties: thermal hysteresis (TH) and ice recrystallization inhibition (IRI). TH allows the IBP to lower the freezing point, meanwhile the IRI property prevents the growth of large ice crystals at the expense of smaller ones. (6)
TmAFP is an antifreeze protein from the insect Tenebrio molitor. Is a small protein (8.4 kDa; 7) which consists of beta helices, numerous disulfide cross-links and threonine side chains on their ice binding faces. (8)
In general, insect AFPs have specific activities 10–100 times greater than those of fish AFPs, even at low concentrations (9). TmAFP has high TH activity of 5.2°C when present at a concentration of 2.7 mg/ml (9). This AFP bonds to the basal plane of the ice crystal blocking ice crystal growth along the c-axis and protecting the crystal from growth down to lower freezing temperatures (10).
References
(1) Xiang H, Yang X, Ke L, Hu Y. The properties, biotechnologies, and applications of antifreeze proteins. Int J Biol Macromol. 2020 Jun 15;153:661–75.
(2) Mahatabuddin S, Tsuda S. Applications of antifreeze proteins: Practical use of the quality products from Japanese fishes. In: Advances in Experimental Medicine and Biology. Springer New York LLC; 2018. p. 321–37.
(3) Balamurugan S, Ann JS, Varghese IP, Murugan SB, Harish MC, Kumar SR, et al. Heterologous expression of Lolium perenne antifreeze protein confers chilling tolerance in tomato. J Integr Agric. 2018 May 1;17(5):1128–36.
(4) Bredow, M., Vanderbeld, B., & Walker, V. K. (2017). Ice-binding proteins confer freezing tolerance in transgenic Arabidopsis thaliana. Plant Biotechnology Journal, 15(1), 68–81.
(5) Bredow M, Walker VK. Ice-binding proteins in plants. Front Plant Sci. 2017;8(December):1–15.
(6) Davies, P. L. (2014). Ice-binding proteins: A remarkable diversity of structures for stopping and starting ice growth. Trends in Biochemical Sciences, Vol. 39, pp. 548–555.
(7) Liou YC, Tocilj A, Davies PL, Jia Z. Mimicry of ice structure by surface hydroxyls and water of a β-helix antifreeze protein. Nature. 2000 Jul 20;406(6793):322–4.
(8) Yue CW, Zhang YZ. Cloning and expression of Tenebrio molitor antifreeze protein in Escherichia coli. Mol Biol Rep. 2009;36(3):529–36.
(9) Bar, M., Bar-Ziv, R., Scherf, T., & Fass, D. (2006). Efficient production of a folded and functional, highly disulfide-bonded β-helix antifreeze protein in bacteria. Protein Expression and Purification, 48(2), 243–252.
(10) Middleton, A. J., Marshall, C. B., Faucher, F., Bar-Dolev, M., Braslavsky, I., Campbell, R. L., … Davies, P. L. (2012). Antifreeze protein from freeze-tolerant grass has a beta-roll fold with an irregularly structured ice-binding site. Journal of Molecular Biology, 416(5), 713–724.
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