Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors
Abstract Miraculin, derived from the miracle fruit (Synsepalum dulcificum), is a taste-regulating protein that interacts with human sweet-taste receptors and transforms sourness into sweet taste. Since miracle fruit is cultivated in West Africa, mass production of miraculin is limited by regional an...
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2020-08-01
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doaj-6105ad3d82f74da182f0c7a620853d3e2020-11-25T02:42:14ZengSpringerOpenAMB Express2191-08552020-08-011011710.1186/s13568-020-01079-3Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactorsYun-Ji Park0Jong-Eun Han1Hyoshin Lee2Yu-Jin Jung3Hosakatte Niranjana Murthy4So-Young Park5Department of Horticulture, Division of Animal, Horticultural and Food Sciences, Chungbuk National UniversityDepartment of Horticulture, Division of Animal, Horticultural and Food Sciences, Chungbuk National UniversityDepartment of Forest Genetic Resources, National Institute of Forest ScienceDepartment of Horticultural Life Science, Hankyong National UniversityDepartment of Horticulture, Division of Animal, Horticultural and Food Sciences, Chungbuk National UniversityDepartment of Horticulture, Division of Animal, Horticultural and Food Sciences, Chungbuk National UniversityAbstract Miraculin, derived from the miracle fruit (Synsepalum dulcificum), is a taste-regulating protein that interacts with human sweet-taste receptors and transforms sourness into sweet taste. Since miracle fruit is cultivated in West Africa, mass production of miraculin is limited by regional and seasonal constraints. Here, we investigated mass production of recombinant miraculin in carrot (Daucus carota L.) callus cultures using an air-lift bioreactor. To increase miraculin expression, the oxidative stress-inducible SWPA2 promoter was used to drive the expression of miraculin gene under various stress treatments. An 8 h treatment of hydrogen peroxide (H2O2) and salt (NaCl) increased the expression of miraculin gene by fivefold compared with the untreated control. On the other hand, abscisic acid, salicylic acid, and methyl jasmonate treatments showed no significant impact on miraculin gene expression compared with the control. This shows that since H2O2 and NaCl treatments induce oxidative stress, they activate the SWPA2 promoter and consequently up-regulate miraculin gene expression. Thus, the results of this study provide a foundation for industrial-scale production of recombinant miraculin protein using transgenic carrot cells as a heterologous host.http://link.springer.com/article/10.1186/s13568-020-01079-3Agrobacterium-mediated transformationAlternative sweetenerCarrot callus culturesMiraculinPlant cell cultureRecombinant protein |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yun-Ji Park Jong-Eun Han Hyoshin Lee Yu-Jin Jung Hosakatte Niranjana Murthy So-Young Park |
spellingShingle |
Yun-Ji Park Jong-Eun Han Hyoshin Lee Yu-Jin Jung Hosakatte Niranjana Murthy So-Young Park Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors AMB Express Agrobacterium-mediated transformation Alternative sweetener Carrot callus cultures Miraculin Plant cell culture Recombinant protein |
author_facet |
Yun-Ji Park Jong-Eun Han Hyoshin Lee Yu-Jin Jung Hosakatte Niranjana Murthy So-Young Park |
author_sort |
Yun-Ji Park |
title |
Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors |
title_short |
Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors |
title_full |
Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors |
title_fullStr |
Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors |
title_full_unstemmed |
Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors |
title_sort |
large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors |
publisher |
SpringerOpen |
series |
AMB Express |
issn |
2191-0855 |
publishDate |
2020-08-01 |
description |
Abstract Miraculin, derived from the miracle fruit (Synsepalum dulcificum), is a taste-regulating protein that interacts with human sweet-taste receptors and transforms sourness into sweet taste. Since miracle fruit is cultivated in West Africa, mass production of miraculin is limited by regional and seasonal constraints. Here, we investigated mass production of recombinant miraculin in carrot (Daucus carota L.) callus cultures using an air-lift bioreactor. To increase miraculin expression, the oxidative stress-inducible SWPA2 promoter was used to drive the expression of miraculin gene under various stress treatments. An 8 h treatment of hydrogen peroxide (H2O2) and salt (NaCl) increased the expression of miraculin gene by fivefold compared with the untreated control. On the other hand, abscisic acid, salicylic acid, and methyl jasmonate treatments showed no significant impact on miraculin gene expression compared with the control. This shows that since H2O2 and NaCl treatments induce oxidative stress, they activate the SWPA2 promoter and consequently up-regulate miraculin gene expression. Thus, the results of this study provide a foundation for industrial-scale production of recombinant miraculin protein using transgenic carrot cells as a heterologous host. |
topic |
Agrobacterium-mediated transformation Alternative sweetener Carrot callus cultures Miraculin Plant cell culture Recombinant protein |
url |
http://link.springer.com/article/10.1186/s13568-020-01079-3 |
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