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High-sweetness sweet protein gene and synthesis method thereof

A synthetic method and gene technology, applied in the field of bioengineering, can solve the problem of sweetness and sweetness of sweet proteins

Inactive Publication Date: 2009-11-04
刘松财 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention provides a high-sweetness sweet protein gene, which is a modification of the Brazzein gene, and solves the problem that the sweet protein expressed by genetic engineering has no sweet taste or low sweetness

Method used

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  • High-sweetness sweet protein gene and synthesis method thereof
  • High-sweetness sweet protein gene and synthesis method thereof
  • High-sweetness sweet protein gene and synthesis method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. SOE-PCR synthesis of NBrazzein gene:

[0035] For the first PCR, add each component in sequence according to Table 2-1 and perform PCR reaction. Reaction conditions: pre-denaturation at 94°C for 5 minutes, 55°C for 15 minutes, and extension at 72°C for 15 minutes after one cycle.

[0036] Table 2-1 Primary PCR reaction system

[0037]

[0038] For the second PCR, add each component in sequence according to Table 2-2 and perform PCR reaction. Reaction conditions: 94°C pre-denaturation for 3 minutes, 94°C for 30s, 55°C for 30s, 72°C for 60s, 30 cycles, 72°C for 10 minutes. The primary PCR product was used as a template, and then PCR was carried out using P1 and P6, P3 and P8 as primers respectively.

[0039] Table 2-2 Primary PCR reaction system

[0040]

[0041]

[0042] Three-time PCR: The product of the second PCR PCR is used as a template, and the third PCR is performed with P1 and P8. The system is shown in Table 2-3. The reaction conditions were pre-d...

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Abstract

The invention discloses a high-sweetness sweet protein gene and a synthesis method thereof. Codons preferred by yeast are adopted to design a gene sequence by knocking out the fist amino acid at an amino terminal, turning 29-bit aspartic acid Asp into lysine Lys, turning 41-bit glutamic acid Glu into lysine Lys, designing four pairs of primers according to the characteristics of pGAPZ alphaA multiple clone sites and the restriction enzyme property of a Brazzein gene, inducing an Xbak restriction enzyme recognition site at an upstream primer, inducing an Xhol restriction enzyme recognition site at a downstream primer and adopting an SOE-PCR method to obtain a Brazzein gene product with the full length of 188bp. The sweetness of modified sweet protein is 2 to 6 times of the sweetness of the original Brazzein gene.

Description

Technical field: [0001] The invention provides a high-sweetness sweet protein gene, and also provides a synthesis method of the sweet protein gene, which belongs to the technical field of bioengineering. Background technique [0002] Sweetener refers to a flavoring agent that can impart sweetness to food, and is mainly used as an additive or pharmaceutical excipient in food processing. The sweeteners in the early years were all derived from nature, and monosaccharides and oligosaccharides with various structures are the most important type, so it is often necessary to add a large amount to food to obtain sufficient sweetness, and it can also induce diabetes, Obesity and dental caries, etc., so people's demand for non-sugar sweeteners is increasing. Sugar alcohol sweeteners have low sweetness and can easily cause abdominal discomfort, flatulence and even diarrhea after eating. Although sugar ligand sweeteners have higher sweetness, their sweetness appears slowly in the mouth...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/10C12N15/63C07K14/415
Inventor 刘松财张丽萍王长远史勇唐丹
Owner 刘松财
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