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KMID : 0380020210360040285
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Korean Journal of Biotechnology and Bioengineering
2021 Volume.36 No. 4 p.285 ~ p.295
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Over-expression and Host-specific Algicidal Effect of Virus-like Particles from Capsid Gene of HcRNAV34 Virus Infecting against a Harmful alga, Heterocapsa circularisquama
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Seo Sun-Il
Ryu Jae-Won
Kim Si-Wouk
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Abstract
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Harmful algal blooms (HABs) is a rapid increase or accumulation in the population of harmful microalgae in freshwater and marine aquatic system, and can harm the health of the environment, plants or animals. Conventional chemical and physical techniques to mitigate HABs such as chlorination, ozonation, ultrasonic treatment, and clay-based flocculation are limited by the low efficiency and risk of destroying the aquatic ecosystems. Therefore, host-specific virus-like particles (VLPs) as an alternative biological method for the efficient control of harmful microalgae that can be combined with chemically synthesized algicidal compound has been successfully performed in this lab. This system was efficient for algicidal activity but also involved several significant disadvantages such as difficult purification and less production rate. To overcome these issues, we attempted to develop a low-cost, constitutive overexpression, and easy purification system of viral capsid protein to obtain large amounts VLPs. The capsid protein of HcRNAV34 as a singlestranded RNA virus that infects the toxic dinoflagellate, Heterocapsa circularisquama, was successfully expressed in E. coil without inducer. The maximal condition for production of HcRNAV34 capsid protein in the E. coil BL21(DE3) was optimized, and then finally 3.25 g/ℓ (wet weight) cell mass was obtained from 30 L reactor containing M9 medium at 37oC for 20 h. The expressed insoluble capsid protein was partially purified by low speed centrifugation and labelled with green fluorescent FITC. The resulting protein was visualized on the cell surface of H. circularisquama 9433 and 92-1, the hosts of native HcRNAV34 virus. In addition, HcRNAV34 VLPs absorbed with TD49 as algicidal substance showed a high algicidal effect against H. circularisquama 9433 and 92-1 in a host-specific manner. This host target-specific VLPs can be used for the management of HABs in aquatic ecosystems.
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KEYWORD
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harmful algal bloom, Heterocapsa circularisquama, HcRNAV virus, capsid protein, virus-like particles
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