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Whey protein film and preparation method thereof

A technology of whey protein and film-forming liquid, which is applied in the field of whey protein film and its preparation, and achieves the effect of important application value

Inactive Publication Date: 2013-10-16
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above shows that recombinant ana-LOX has the potential to be used as a cross-linking agent to prepare packaging films, but the research in this area has not been reported yet

Method used

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  • Whey protein film and preparation method thereof
  • Whey protein film and preparation method thereof
  • Whey protein film and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Cloning of LOX gene from Anabaena sp. PCC 7120

[0023] Anabaena sp. PCC 7120 cells were collected by centrifugation, and the genomic DNA of Anabaena sp. was extracted with the Shanghai Sangon Genomic DNA Extraction Kit.

[0024] Two primers were designed according to the genome sequence published by NCBI (NC_003267):

[0025] Upstream primer: 5'-GGAGTGTCTGGTGCC-3' (SEQ ID NO.3)

[0026] Downstream primer: 5'-CTAAATGTTGATACTCATCAT-3' (SEQ ID NO.4)

[0027] In a 50 μl system, the final concentration of each primer is 1 μM, the final concentration of dNTPs is 0.2 mM, 10 ng of Anabaena genomic DNA, and 2 U of Pfu DNA polymerase. The amplification program was 94°C for 3min; 30×(94°C for 30s, 59°C for 50s, 72°C for 40s); 72°C for 10min. Agarose gel electrophoresis, gel cutting, recovery by Shanghai Sangon kit, connection of the recovered PCR product with TaKaRa pMD19-T vector, transformation of E.coli DH5α, and spreading on LB plates containing IPTG, X-gal, and...

Embodiment 2

[0028] Example 2: Construction of anabaena sp. PCC 7120 ana-LOX gene expression vector

[0029] According to the obtained LOX gene sequence, two primers were designed, the upstream primer plus the SacI recognition sequence, and the downstream primer plus the XhoI recognition sequence (the underlined part is the restriction enzyme recognition sequence):

[0030] Upstream primer ana-LOX-F: 5′-CGC GAGCTC GGAGTGTCTGGTGCC-3' (SEQ ID NO.5)

[0031] Downstream primer ana-LOX-R: 5′-CCG CTCGAG CTAAATGTTGATACTCATCAT-3' (SEQ ID NO. 6)

[0032]Add each component according to the following PCR system to amplify the LOX gene:

[0033]

[0034] The PCR program was 94°C for 2min; 30×(94°C for 45s; 58°C for 50s; 72°C for 4min); 72°C for 10min.

[0035] Purify the PCR product with Shanghai Sangon PCR Product Purification Kit, add SacI, XhoI double enzyme digestion, inactivation, ethanol precipitation, ddH 2 O was redissolved, ligated with an appropriate amount of vector pET-23a digest...

Embodiment 3

[0036] Example 3: Expression of Anabaena sp.PCC 7120 LOX gene in Escherichia coli

[0037] Transform the Escherichia coli expression host strain BL21(DE3)pLysS (purchased from Novagen) containing the ana-LOX expression plasmid pET-23a-ana-LOX, pick small colonies after culturing at 37°C for 10-11 hours, and insert them into the Penicillin 50ml LB liquid medium, 70-90rpm, culture overnight at 30°C, take the seed solution according to the volume ratio of 1:40 and add it to 100ml LB liquid medium containing ampicillin, shake at 35°C and 180rpm for 2-3 hours until the OD600 is about 0.6 Then add IPTG (final concentration 100μg / ml) to induce. After 5 hours, the cells were collected by centrifugation.

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Abstract

The invention belongs to the field of food industrial biotechnology, and relates to a whey protein film and a preparation method thereof. The whey protein film is prepared by adding whey protein and glycerol into purified water to prepare film-forming liquid, heating the film-forming liquid in 45-50 DEG C water bath for 0.5-2h, cooling to room temperature, adding recombinant lipoxygenase and linoleic acid, ultrasonically degassing, pouring into a glass culture dish covered with a plastic film, reacting for 2-3h at room temperature (20-25 DEG C), drying at 50-55 DEG C for 36-48h, and uncovering the film. With the whey protein as a based material and the glycerol as a plasticizer, the whey protein film prepared by adding the recombinant anabaena lipoxygenase as a crosslinking agent is green, edible and good in tensile strength, transparency, water absorption and plasticity.

Description

technical field [0001] The invention belongs to the field of food industry biotechnology, and relates to a whey protein film and a preparation method thereof. Background technique [0002] While plastic products have brought great convenience to society since their creation, they have also brought great threats to the living environment of human beings. This kind of packaging made of high molecular chemical polymer materials is not only difficult to degrade, but also produces polluting gases due to combustion. With the enhancement of people's awareness of environmental protection, the use of natural green materials to prepare edible packaging films has become the focus of many researchers. Edible packaging materials are made of natural edible substances, such as proteins, polysaccharides (starch, cellulose), etc., and have a porous network structure formed by intermolecular cross-linking. They are edible, easy to degrade, and have a certain Pack protection features. [00...

Claims

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Application Information

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IPC IPC(8): C08J5/18C08J3/24C08L89/00C08K5/053C08K5/09C12N15/53C12N15/70C12N9/02
Inventor 陆兆新张充吕凤霞别小妹赵海珍
Owner NANJING AGRICULTURAL UNIVERSITY
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