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Method for promoting quality of fish scale gelatin

A technology for fish scale gelatin and glue quality, which is applied in the directions of chemical instruments and methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of poor quality and low content of gelatin glue such as melting temperature and strength, etc., Achieve high-value utilization, improve gelatin quality, and achieve the effect of green and sustainable development

Active Publication Date: 2019-03-22
JIANGXI NORMAL UNIV
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Problems solved by technology

However, compared with gelatin derived from mammals, scale gelatin (SG) has relatively low content of imino acid-hydroxyproline, so that gelatin does not have enough hydroxyl groups to form hydrogen bonds with water to stabilize the triplet helix of gelatin. structure, resulting in gelatin melting temperature and strength and other properties inferior to mammalian gelatin

Method used

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  • Method for promoting quality of fish scale gelatin
  • Method for promoting quality of fish scale gelatin
  • Method for promoting quality of fish scale gelatin

Examples

Experimental program
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Embodiment 1

[0028] The construction process of recombinant bacteria and the purification method of P4H comprise the following steps:

[0029] (1) Construction of expression P4H plasmid and construction of recombinant bacteria: select Pseudomonas fluorescens as the source bacteria of P4H, check its use of codons, and find that the sequence has 9 scattered arginine codons aga, which are not used by Escherichia coli, Therefore, after replacing the codons at these sites with arginine codons commonly used in Escherichia coli, add XhoI and EcoRI restriction enzymes to both ends of the DNA sequence and delete the stop codon, and then send it directly to Nanjing GenScript Synthesized by Biotechnology Co., Ltd. (the P4H coding gene is synthesized, because the P4H in this strain contains many preferred codons, so the synthesis is more economical). The synthesized DNA fragment and the vector pET-28(a) were digested and purified with XhoI and EcoRI respectively, and ligated under the action of ligase...

Embodiment 2

[0032] Adopting P4H to modify fish phosphorus gelatin, the modification method comprises the following steps:

[0033] (1) To characterize the characteristics of P4H: take the amount of proline converted into hydroxyproline as the detection standard, explore the best reaction pH value, the best reaction temperature and the best reaction system, and determine the best reaction conditions enzyme activity. Reaction system for converting proline into hydroxyproline: the P4H crude enzyme obtained after culturing the above-mentioned recombinant bacteria for crushing, each reaction system (250 μL) contains proline at a final concentration of 20 mmol / L, and the final concentration is 40 mmol / L L of α-ketoglutaric acid, ferrous sulfate with a final concentration of 4 mmol / L, and vitamin C with a final concentration of 8 mmol / L. The results showed that the optimum reaction pH value was 5.5, the optimum reaction temperature was 26°C, and the optimum reaction buffer system was Tris / HCl b...

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Abstract

The invention discloses a method for promoting quality of fish scale gelatin. L-proline cis-4-hydroxylase (P4H) is effectively expressed in escherichia coli and is used for modifying fish scale gelatin, so as to promote the quality of fish scale gelatin and expand the application scope thereof in the market.

Description

technical field [0001] The invention relates to a method for efficiently expressing proline hydroxylase (L-proline cis-4-hydroxylase, P4H) by using genetic engineering technology and using it to modify gelatin derived from fish scales to improve its quality. Background technique [0002] Edible gelatin (gelatin) is a hydrolyzed product of collagen, a fat-free, high-protein, cholesterol-free, and a natural nutritional food thickener. As a powerful protective colloid, gelatin has strong emulsifying power, which can effectively inhibit the coagulation reaction caused by gastric acid after protein such as milk and soy milk enters the stomach, and is beneficial to improve food digestion. In addition to being widely used in the food field, gelatin also plays its special functions in the fields of medicine, cosmetics, and photography. At present, most of the gelatin used in industry comes from mammals, such as pig skin, cowhide, pig bone, and beef bone. However, in recent years, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/70C12P21/00C07K14/78
CPCC12N9/0071C12N15/70C07K14/78Y02A40/81
Inventor 陈雪岚涂宗财李双黄明珠
Owner JIANGXI NORMAL UNIV
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