Part:BBa_K3612008
Lolium perenne antifreeze protein (LpIRI3) - P. nigrifaciens
LpIRI3 is an antifreeze protein from the grass Lolium perenne, and given that has higher IRI activity compared to LpAFP, our team chose it as one of the AFPs to be expressed first in E. coli and later in Pseudoalteromonas nigrifaciens. The expression in the latter organism will allow us to produce LpIRI3 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, 4, 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)
LpIRI3 is an antifreeze protein from the ryegrass Lolium perenne. LpIRI3 is predicted to have the same structure of LpAFP, which means eight helical beta loops in its secondary structure. Moreover, given its absorption to the primary prism and basal planes,it is predicted to be a hyperactive AFP. (4)
LpIRI3 has high IRI activity but low TH activity, indicating that plants rely on restricting ice crystal growth as a primary survival strategy. Although LpIRI3 is not well known as it is LpAFP, recent studies demonstrate that LpIRI3 has higher activity than the latter one in E. coli and in transgenic plants, significantly enhancing freeze survival. (4)
Although LpAFP expression has not been tested in the marine bacteria of Pseudoalteromonas genus, many studies have proposed it as a non-conventional system for recombinant protein production (7-9). Hence, our team considers that it would be an innovative and useful chassis for AFP production in the conditions of the Andean regions.
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) Sannino F, Giuliani M, Salvatore U, Apuzzo GA, de Pascale D, Fani R, et al. A novel synthetic medium and expression system for subzero growth and recombinant protein production in Pseudoalteromonas haloplanktis TAC125. Appl Microbiol Biotechnol. 2017 Jan 27;101(2):725–34.
(8) Yu ZC, Tang BL, Zhao DL, Pang X, Qin QL, Zhou BC, et al. Development of a cold-adapted Pseudoalteromonas expression system for the Pseudoalteromonas proteins intractable for the Escherichia coli system. PLoS One. 2015;10(9):1–14.
(9) Parrilli E, Tutino ML. Psychrophiles: From Biodiversity to Biotechnology: Second Edition. Psychrophiles From Biodivers to Biotechnol Second Ed. 2017;1–685.
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