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Method for improving stability of liquid aminopeptidase and application of aminopeptidase

An aminopeptidase and stability technology, applied in the field of enzyme preparations and food additives, can solve the problems of limiting the application of protein hydrolyzate, and achieve the effect of easy expansion of production and good application prospects

Inactive Publication Date: 2012-05-23
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Protein hydrolysis produces peptides and a small amount of amino acids, whose solubility, thermal stability, and nutritional properties are greatly improved, but the hydrolyzate contains bitter peptides, which generally have single or multiple hydrophobic amino acids at the amino terminal Polypeptide chains, sometimes with hydrophobic amino acids at the carboxyl end, the bitter taste limits the application of protein hydrolysates

Method used

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  • Method for improving stability of liquid aminopeptidase and application of aminopeptidase
  • Method for improving stability of liquid aminopeptidase and application of aminopeptidase
  • Method for improving stability of liquid aminopeptidase and application of aminopeptidase

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Embodiment 1

[0044] Centrifuge the Bacillus subtilis Zj016 fermentation broth at 4°C for 10 min at 10,000 rpm to collect 3.5L of the fermentation broth, add (NH 4 ) 2 SO 4670g (stirring while adding), 4°C, 10000rpm centrifuge for 10min to collect 3.6L of clarified liquid, select 50kDa ultrafiltration membrane to concentrate to 1.4L, flow rate 30-35mL / min; select 30kDa ultrafiltration membrane to desalinate 1.4L, The flow rate is 40mL / min; add 3.3L of pre-cooled absolute ethanol to the desalination solution, and after 10min, centrifuge at 4°C and 8000rpm for 10min to collect the precipitate, dissolve the precipitate, add sodium benzoate as a stabilizer, and control the quality of sodium benzoate in the enzyme solution The concentration is 0.1%, and the liquid aminopeptidase is obtained. Soybean protein isolate is mixed with a 6% solution, placed in a constant temperature water bath at 85°C for 15 minutes, cooled to adjust the pH to 8.5, and the amount of enzyme added E / S=3.25×10 5 U / g, t...

Embodiment 2

[0046] Centrifuge the Bacillus subtilis Zj016 fermentation broth at 4°C for 10 min at 10,000 rpm to collect 2.5L of the fermentation broth, add (NH 4 ) 2 SO 4 470g (stirring while adding), 4°C, 10000rpm centrifuge for 10min to collect 2.5L of clarified liquid, select 50kDa ultrafiltration membrane to concentrate to 0.85L, flow rate 30-35mL / min; select 30kDa ultrafiltration membrane to desalinate 0.85L, The flow rate is 40mL / min, add 1.8L of pre-cooled absolute ethanol to the desalination solution, and after 10min, centrifuge at 4°C and 8000rpm for 10min to collect the precipitate, dissolve the precipitate, and add 0.2μmol / L Co 2+ The cofactor is liquid aminopeptidase. Soybean protein isolate is mixed with a 6% solution, placed in a constant temperature water bath at 85°C for 15 minutes, cooled to adjust the pH to 8.5, and the amount of enzyme added E / S=2.5×10 5 U / g, the content of free amino acids in the protein hydrolyzate hydrolyzed at 50°C for 4h is 64.72mg / mL, macromole...

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Abstract

The invention discloses a method for improving the stability of liquid aminopeptidase and an application of the aminopeptidase, which belong to the technical field of enzymic preparations and food additives. The invention relates to an extracting process of bacillus subtilis Zj016 liquid aminopeptidase to improve the stability of the liquid aminopeptidase and an application of the aminopeptidase in deep hydrolysis of soy isolated protein. The bacillus subtilis Zj016 liquid aminopeptidase is centrifuged, clarified, concentrated, desalinized, deposited by alcohol and redissolved to obtain the liquid aminopeptidase; the liquid aminopeptidase can keep stable by controlling the pH condition and / or the additive; the aminopeptidase can carry out deep hydrolysis on a substrate of the soy isolatedprotein to generate a small peptide and amino acid with higher content and has a combined application potential with a plurality of incision enzymes, thereby having wide application prospect in food processing of protein raw materials.

Description

technical field [0001] A method for improving the stability of food-grade bacillus subtilis liquid aminopeptidase and the application of the enzyme. Using the fermented liquid of Bacillus subtilis, the liquid aminopeptidase is obtained by centrifugation, clarification, concentration by ultrafiltration, desalination, and alcohol precipitation. The stability is improved by adding stabilizers or cofactors, and the application in food and additives is studied. The invention belongs to the technical field of enzyme preparations and food additives. Background technique [0002] Enzyme preparations have been widely used in various fields such as food, brewing, pharmaceuticals, organic acids, starch sugar, feed industry, textiles, leather, detergents and health products, and the application fields are becoming more and more extensive. Food is closely related to people's lives, so the application value of enzymes in the food industry is becoming more and more important. At present, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/56C12P21/06C12R1/125
Inventor 田亚平吕广林高新星李欣艳
Owner JIANGNAN UNIV
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