Method for preparing thermal reversible starch based gelatin with controllable gelatin strength by enzyme method

A gel strength, starch-based technology, applied in the field of starch modification, can solve the problems of difficult application of starch gel products, uncontrollable, high starch gel strength, and achieve a wide range of applications, good temperature stability, high gelation the effect of strength

Inactive Publication Date: 2020-01-07
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the many defects of native starch gel, the strength of starch gel is too high and uncontrollable, which makes starch gel products difficult to apply

Method used

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  • Method for preparing thermal reversible starch based gelatin with controllable gelatin strength by enzyme method
  • Method for preparing thermal reversible starch based gelatin with controllable gelatin strength by enzyme method
  • Method for preparing thermal reversible starch based gelatin with controllable gelatin strength by enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: the preparation of enzyme

[0041] The artificially synthesized gene fragment whose nucleotide sequence is shown in SEQ ID NO.2 (the amino acid sequence is shown in SEQ ID NO.1) is selected from the pET-28a plasmid to construct a recombinant plasmid, expressed by BL21 expression strain, and fermented by shaking flasks. Centrifuge and precipitate, sonicate the wall to obtain crude enzyme, and finally obtain 4αGTase pure enzyme after nickel column purification (see figure 1 ).

[0042]MLRGAGILLPLFSLPGPHGIGDMGPAAYRFVEFLRDAGQTYWQLLPLNPVLPEYDNSPYSSTSSFAGEPAYISLELLAEQGVLRRSASAEFGAGRADYEAARAYKVKIIAEAERPKDLDEFVGRNEWLEDYALYSALREYFGRPWVEWPRELRDRDPAALRQWREKLRRRIELEYLLQYFFWTQWRELKRHANSLGIFLIGDLPIYLSLDSADVWAHRELFKLDAEGRPLYVAGVPPDYFSPTGQLWGNPVYDWDAMRKEGFRWWLDRLRYTLSVFDYVRLDHFRGYAAYWEVPAGEKTAVNGRWVPAPGGELLERARDELGGLKIIAEDLGYITPDVVELRDRFGLPGMRVLQFAWDGNPANEHKPHNHVKNSVVYTGTHDNNTIVGWFFREASPKARREALEYMGRRSAKDIHWAFIRLAYFSVADVAVVPMQDFMGLGPEARINRPGTRGGNWVWRMTELPGRALARRIRRLARLYGR ...

Embodiment 2

[0043] Example 2: Properties of Enzymes

[0044] (1) The optimal reaction temperature of the enzyme

[0045] The disproportionation activity of 4αGTase was measured at different temperatures (50°C-90°C) to determine the optimum temperature. The optimum reaction temperature of 4-α-glycosyltransferase is 50~80℃.

[0046] Table 1 Optimum reaction temperature of 4αGTase

[0047]

[0048] (2) The optimal reaction pH of the enzyme

[0049] The buffer in the 4αGTase disproportionation assay method was replaced by buffers with different pH values ​​(3.0-10.0), and the activity of 4αGTase was measured at 75°C to investigate the optimum pH of the enzyme. The optimum reaction pH is 5.0-8.0.

[0050] Table 2 Determination of optimum pH of 4αGTase

[0051]

[0052]

[0053] (3) Temperature stability of the enzyme

[0054] Incubate at different temperatures (50°C-90°C), take out part of the enzyme solution every 1 hour, cool down rapidly, and measure the remaining total enzym...

Embodiment 3

[0057] Embodiment 3: the impact of starch type on the performance of starch-based gel

[0058] (1) The starch with a mass concentration of 6% is dissolved in a buffer solution for slurry preparation, and is completely gelatinized at 120°C.

[0059] (2) Lower the temperature to 70°C, adjust the pH to 6.0, add 4-α-glycosyltransferase, react for 1-12 hours, then inactivate the enzyme, dry, and grind to obtain modified starch;

[0060] (3) Dissolving the obtained modified starch, heating and stirring at 70-90° C., and obtaining the modified starch gel after solidification. The starch mainly includes potato starch, corn starch, tapioca starch, wheat starch, pea starch and rice starch, and the results are shown in Table 4.

[0061] Table 4 The impact of starch types on the properties of modified starch

[0062] Starch type Potato starch corn starch Cassava starch wheat starch pea starch rice starch Conversion rates(%) 92.61 32.56 88.79 21.08 54.24 ...

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Abstract

The invention discloses a method for preparing thermal reversible starch based gelatin with controllable gelatin strength by an enzyme method, and belongs to the technical field of starch modification. The method comprises the steps of cloning 4-alpha-Glyt (Tu alpha GT) in Thermoproteus uzoniensis (Thermoproteus uzoniensis), performing escherichia coli heterologous expression, performing enzymology nature characterization, and determining that the 4-alpha-Glyt (Tu alpha GT) is 4 alpha GT, the enzyme has good temperature stability and substrate tolerance, high-concentration starch can be catalyzed at high temperature for modification, and the side chain of branched chain starch can be prolonged. According to the method for preparing thermal reversible starch based gelatin with controllablegelatin strength by an enzyme method, the starch is used as the raw material, an appropriate amount of the 4-alpha-Glyt is added, an appropriate additive quantity and appropriate action time are selected, and the high strength anabiotic resisting starch based gelatin can be prepared.

Description

technical field [0001] The invention relates to a method for enzymatically preparing thermoreversible starch-based gel with controllable gel strength, belonging to the technical field of starch modification. Background technique [0002] Starch is a pure natural raw material with a wide range of sources and low price. Starch colloid produced by using starch as raw material is widely used in many industries such as food and chemical industry. Starch gel is a polymer compound, which is a transparent or translucent gel with certain elasticity and hardness formed after starch gelatinization. In the food industry, compared with other food colloids, starch colloids are easier to be digested and absorbed by the human body, less prone to adverse reactions such as flatulence and nephrotoxicity, and can be added to infant food, with broad market prospects. Among other industries, starch colloids have the advantages of light weight, no corrosion, no pollution and low cost. However, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/18C12P19/04C08J3/075C08L3/02A23L29/30
CPCA23L29/30C08J3/075C08J2303/02C12P19/04C12P19/18
Inventor 柏玉香王禹金征宇
Owner JIANGNAN UNIV
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