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Method for producing gamma-aminobutyric acid by using banana biological enzyme method

A technology of aminobutyric acid and bio-enzyme method, which is applied in the field of producing γ-aminobutyric acid by banana bio-enzyme method, can solve complex processing problems, and achieve the effects of simple production process, specific reaction and convenient operation

Active Publication Date: 2013-07-17
CATCH BIO SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method can improve the processing depth of fruits and vegetables and increase the added value of fruits and vegetables, but the processing is complicated

Method used

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  • Method for producing gamma-aminobutyric acid by using banana biological enzyme method
  • Method for producing gamma-aminobutyric acid by using banana biological enzyme method
  • Method for producing gamma-aminobutyric acid by using banana biological enzyme method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Prepare the extraction solution: 0.05mol / L Na 2 HPO 4 / NaH 2 PO 4 , 0.15mol / L NaCl, 0.2mmol / L vitamin B 6 Or pyridoxal phosphate PLP, 30mg / L CaCl 2 , pH 5.7;

[0032] (2) Chop and grind the ripe whole banana (with belt and pulp), add 2L of extraction solution to each 1kg of banana, stir well, and stand at room temperature for extraction for 4 hours;

[0033] (3) After centrifugation at 7000rpm for 10 minutes, the supernatant is glutamic acid decarboxylase solution;

[0034] (4) Add 15g of glutamic acid or sodium glutamate to the glutamic acid decarboxylase solution, control the pH value at 4.5-5.5, stir at 150r / min for 2-3 hours at 37°C, and then add 3-3 5g of glutamic acid or sodium glutamate, the pH value is controlled at 5.0-5.5, and this is repeated until the enzyme activity disappears. Here, the enzyme activity is measured using the Berthelot reaction principle;

[0035] (5) After the reaction is completed, add 1% (w / v) ...

Embodiment 2

[0040] (1) Prepare the extraction solution: 0.08mol / L Na 2 HPO 4 / NaH 2 PO 4 , 0.15mol / L NaCl, 0.3mmol / L vitamin B 6 Or PLP, 30mg / L CaCl 2 , pH 5.7;

[0041] (2) Chop and grind the ripe whole banana, add 1L of extract solution to every 1kg of banana, stir evenly, and stand at room temperature for extraction for 4 hours;

[0042] (3) After centrifugation at 7000rpm for 10 minutes, the supernatant is glutamic acid decarboxylase solution;

[0043] (4) Add 500 mL of 20 g / L glutamic acid or sodium glutamate solution with a pH value of 4.5 to 5.5 to the glutamic acid decarboxylase solution, and stir at 150 r / min at 37°C for 1 to 2 hours, and then supplement every 1 hour. Add 3-5g of glutamic acid or sodium glutamate, control the pH value at 5.0-5.5, and repeat this until the enzyme activity disappears;

[0044] (5) After the reaction is completed, add 1% (w / v) activated carbon and stir at 37°C and 100r / min for 30min to decolorize;

[0045] ...

Embodiment 3

[0047] (1) Prepare the extraction solution: 0.05mol / L Na 2 HPO4 / NaH 2 PO 4 , 0.15mol / L NaCl, 0.3mmol / L vitamin B 6 Or PLP, 30mg / L CaCl 2 , pH 5.5;

[0048] (2) Chop and grind the ripe whole banana, add 1.5L of extracting solution to every 1kg of banana, stir evenly, and stand at room temperature for extraction for 4 hours;

[0049] (3) After centrifugation at 7000rpm for 8 minutes, the supernatant is glutamic acid decarboxylase solution;

[0050] (4) Add 10g of glutamic acid to the glutamic acid decarboxylase solution, the pH value is 5.0-5.5, at 37°C, stir at 120r / min for 1-2h, then add 3-5g of glutamic acid every 1h, the pH The value is controlled at 5.0-5.5, and this is repeated until the enzyme activity disappears;

[0051] (5) After the reaction is completed, add 1% (w / v) activated carbon and stir at 37°C and 120r / min for 60min to decolorize;

[0052] (6) Filtration, rotary steaming at 55°C until a small amount of crystals precipitat...

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Abstract

The invention discloses a method for producing gamma-aminobutyric acid by using a banana biological enzyme method, which comprises the following steps of: cutting up and grinding bananas, adding 1-3L of leaching liquor into every 1 kg of bananas, and after the obtained mixture is uniformly stirred, carrying out standing extraction on the obtained mixture for 3-5 h at a temperature of 4-25 DEG C; carrying out centrifugation on the obtained product for 5-10 min at a speed of 5000-7000 rpm so as to obtain glutamic acid decarboxylase crude-enzyme; adding 30-50 g of glutamic acid and sodium glutamate with pH of 4.5-6.0 into every 1L of enzyme, so that the obtained product can be converted completely in 12-24 h under the condition of 37 DEG C; and adding 0.1-3.0% of active carbon into the obtained product, stirring for 30-60 min, and carrying out filtering, rotary steaming and crystallization on the obtained product so as to obtain 20-40 g of gamma-aminobutyric acid. The method is simple in production process, specific in reaction, low in cost, high in yield, good in purity, short in cycle, less in waste, and environmental-friendly, and leaching liquor can be repeatedly used, therefore, the method is suitable for industrial production.

Description

[0001] technical field [0002] The invention relates to the technical field of biotransformation enzymatic synthesis, in particular to a method for producing gamma-aminobutyric acid using a banana bioenzyme method. [0003] Background technique [0004] Glutamate decarboxylase (Glutamate decarboxylase, GAD) widely exists in prokaryotes and eukaryotes, is an intracellular enzyme, mainly exists in the cytoplasm, and combines with the cofactor pyridoxal phosphate (PLP) to catalyze the conversion of glutamate for GABA and CO 2 . In plants, the C-terminus of GAD has a special calmodulin (CaM) binding region, which can bind to CaM and increase the activity of the enzyme. The molecular weight of banana glutamic acid decarboxylase is 56kD, and the full length of ORF of banana glutamic acid decarboxylase gene cDNA is 1500bp, encoding 499 amino acids. [0005] γ-aminobutyric acid (γ-aminobutyric acid, GABA) molecular formula is C 4 h 9 NO 2 , is a natural non-prote...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00
Inventor 殷志敏王期孙洪露温尧林
Owner CATCH BIO SCI & TECH
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