Industrial applications of plant cell extracts comprising sod enzymes of extremophilic micro-organisms

A plant cell and extract technology, applied in plant cells, medical preparations containing active ingredients, applications, etc., can solve problems such as pollution, restricted use, anti-UV and poor temperature stability

Inactive Publication Date: 2021-06-04
ARTERRA BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, these enzymes are inactive under harsher conditions of temperature, salinity and pH than those mentioned above, as well as under conditions of exposure to UV radiation, which limits their use at an industrial level
[0014] For example, the use of SOD in sunscreen products for skin presents problems with poor UV resistance and temperature stability; enzymes added to health products exhibit problems related to structural stability due to the extremely acidic pH in the gastric environment; and finally, Products intended for the food market are often characterized by high concentrations of salt, which can also negatively affect the enzymatic activity of SOD
[0015] Furthermore, it is known that enzymes produced by the process of isolation and purification from bacteria may be subject to possible contamination with toxic or allergenic compounds of bacterial origin

Method used

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  • Industrial applications of plant cell extracts comprising sod enzymes of extremophilic micro-organisms
  • Industrial applications of plant cell extracts comprising sod enzymes of extremophilic micro-organisms
  • Industrial applications of plant cell extracts comprising sod enzymes of extremophilic micro-organisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Expression of superoxide dismutase (SOD) in tomato cell cultures and characterization of cell-derived extracts belonging to the microorganism Sulfolobus solfataricus.

[0097] To express the gene encoding superoxide dismutase (SsSOD) (SEQ ID NO 1) of the extremophile Sulfolobus solfataricus in tomato cells (MicroTom cell line), the binary vector pBI121 modified with the above gene was transferred to the root Agrobacterium carcinoma (LBA4404 trunk) was used for gene transformation of tomato cotyledon (S. lycopersicum).

[0098] Infected plant tissue is then transferred to a specific medium for callus induction containing the antibiotic kanamycin for selection of transformed cells. After approximately 6 weeks from gene transformation, abundant calli were regenerated, isolated from remaining plant tissue, transferred to fresh medium, and analyzed by semi-quantitative RT-PCR. figure 1 A shows the analysis involving some SsSOD expressing cell lines. Amplification products ...

Embodiment 2

[0159] Expression of SOD in plant cell cultures and characterization of cell-derived extracts belonging to the microbial thermophilic spring-forming archaea.

[0160] Tomato was genetically transformed with the thermophilic springtime archaea SOD (ApSOD) (SEQ ID NO 2) using Agrobacterium tumefanciens as described in Example 1 . Four transformed cell lines were analyzed by RT-PCR using specific primers for ApSOD, and the amplification products were compared with amplified 18S ribosomal RNA obtained in the same reaction (multiplex PCR) ( Figure 4 A). All clones analyzed showed high levels of SOD expression and, as expected, no amplified bands were found in unprocessed tomato cells (wild type, WT) used as a control.

[0161] Western blot analysis indicated that the transgenic cell lines contained detectable amounts of ApSOD protein, whereas in untransformed cell lines, slight hybridization bands were barely visible due to the high similarity between endogenous tomato SOD and ex...

Embodiment 3

[0184] Expression of SOD belonging to the microorganism Deinococcus radiodurans in plant cell cultures and characterization of cell-derived extracts.

[0185] Stable transfer of Deinococcus radiodurans SOD (DrSOD) (SEQ ID NO 3) to tomato by Agrobacterium tumefaciens.

[0186] Calli selected on selective media were analyzed by RT-PCR using specific primers for the SOD transfer sequence. Figure 7 A shows RT-PCR of 5 transgenic cell lines. Amplification products obtained using specific oligonucleotides directed against the DrSOD sequence were normalized to amplified 18S ribosomal RNA obtained in the same reaction (multiplex PCR). All clones analyzed showed high levels of SOD expression. The cell lines were then analyzed in Western blot as Figure 7 In B, the transgenic tomato cell line expresses DrSOD at high levels, whereas a slight band corresponding to endogenous SOD is present in WT cell extracts.

[0187] Cell line 3 extracts containing DrSOD were used to characterize t...

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Abstract

The present invention relates to a composition with an antioxidant activity, comprising a first dry extract derived from plant cell cultures comprising superoxide dismutase of Sulfolobus solfataricus, a second dry extract derived from plant cell cultures comprising superoxide dismutase of Aeropyrum pernix and a third dry extract derived from plant cell cultures comprising superoxide dismutase from Deinococcus radiodurans, wherein said plant cell cultures are cultures of cells belonging to Solanum lycopersicum species.

Description

technical field [0001] The present invention relates generally to the industrial use of extracts derived from plant cell cultures comprising enzymes of extremophiles for the fields of dermocosmetics, biomedicine, nutraceuticals and food, and related compositions and preparations comprising them . Background technique [0002] Oxidation of proteins, DNA, lipids, and other macromolecules known to form living cells is caused by endogenous (metabolic and other cellular processes) or exogenous (exposure to cigarette smoke, ozone, ionizing radiation, heavy metals, and UV radiation) Chemical processes triggered by reactive oxygen species (ROS) or other free radicals. The main ROS with important physiological significance is: superoxide anion (O 2 - ), free radical hydroxyl (OH - ) and hydrogen peroxide (H 2 o 2 ). At moderate concentrations, ROS are involved in the physiological processes of cells, but at higher concentrations, ROS can change the structure of DNA and irrever...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K8/66A23C9/12A01N3/00A23L3/3571A61K38/44A61Q19/08C12N5/04C12N15/82A61K8/9789
CPCA23L3/3571A61K8/66A61K8/9789A61K2800/522A61K2800/524A61K2800/86A61Q19/08A23B4/22C12N15/8257C12N9/0089A23L29/06A23L33/105A23L3/3472A23V2002/00C12Y115/01001
Inventor 斯特凡尼娅·阿切洛法比奥·安朋玛丽亚·加布里埃尔·科卢奇吉安娜·帕尔米耶里埃尼奥·科卡
Owner ARTERRA BIOSCI
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