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Culture medium and method for high-yield production of amylase by ascosphaera apis fermentation

A technology of glomus amylii and a culture medium is applied in the field of culture medium for high-yielding amylase fermented by glosoma melis, can solve the problems of difficulty in industrialization, complicated extraction process and high cost, and achieves good repeatability and high amylase yield. , the effect of short fermentation cycle

Inactive Publication Date: 2014-01-15
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although amylase exists in both animals and plants, it is difficult to industrialize due to the complicated extraction process and high cost

Method used

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  • Culture medium and method for high-yield production of amylase by ascosphaera apis fermentation
  • Culture medium and method for high-yield production of amylase by ascosphaera apis fermentation
  • Culture medium and method for high-yield production of amylase by ascosphaera apis fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Single factor experiment to determine the optimal composition of culture medium

[0020] (1) Seed medium preparation: Peel potatoes, cut into pieces, weigh 200g, boil in water for 30min, filter through 6 layers of gauze, weigh 20g of glucose, add distilled water to 1000mL, pH is natural. Autoclave at 121°C for 20min.

[0021] (2) Preparation of seed fermentation liquid: inoculate Ascomonas melii on potato agar medium, culture at 30°C for 3 days, and activate after it produces spores; Bacteria blocks (0.5×0.5 cm) were inoculated into the seed medium, and cultured for 72 hours at 25°C with a rotation speed of 140 r / min, which was the seed fermentation broth.

[0022] (3) The above seed medium was inoculated into a Erlenmeyer flask (250 mL) containing 50 mL of fermentation medium at a 5% inoculum amount, and cultured at 25 °C for 48 h at a rotational speed of 140 r / min.

[0023] (4) Preparation of enzyme liquid: take the fermentation liquid in step (3) and centrifuge a...

Embodiment 2

[0044] Orthogonal experiment to determine optimal fermentation conditions

[0045] (1) Orthogonal experiment to determine the best medium

[0046] On the basis of the culture medium determined by the single factor experiment, the concentration of each component and the pH value were changed to prepare different culture medium, and the method was the same as that of steps (1) and (2) in Example 1. See Table 6, inoculate 250mL of the same amount of inoculum (5%) into 250mL of culture solution, and incubate at 25°C for 48 hours at a rotational speed of 140 r / min. Then measure the enzyme activity according to the steps (4) and (5) of Example 1. Select the experimental group number with the optimal result of the orthogonal experiment and the highest enzyme production, and carry out the verification experiment. In order to avoid being cumbersome, the preparation of the culture medium, the preparation of the crude enzyme solution, and the enzyme activity assay method in the emb...

Embodiment 3

[0059] Fungal Amylase Fermentation Method

[0060] (1) Preparation of fermented seeds

[0061] (A) Strain and culture medium

[0062] Bacterial species: Glomus spp. Slant medium: PDA medium

[0063] Liquid seed medium: PDB medium.

[0064] (B) Preparation of seed solution

[0065] Transfer the pure Bacillus palisae on the slope to multiple 250mL Erlenmeyer flasks containing 100mL of culture medium, and then at 25°C (lower temperature fermentation culture is also to inhibit contamination), in reciprocating shaking Cultivate at a bed speed of 140 r / min for 72 hours. When the mycelium grows vigorously and the bacterial liquid becomes viscous, it means that the seeds have grown well.

[0066] In addition, antibiotics can be added to the seed solution before inoculation to kill or inhibit the growth of miscellaneous bacteria, so as to obtain pure strains without miscellaneous bacteria.

[0067] (2) Fermentation culture

[0068] Fermentation medium: optimize the orth...

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Abstract

The invention relates to a fungus fermentation culture medium and a fungus fermentation culture technology, and especially relates to a culture medium and a method for high-yield production of amylase by ascosphaera apis fermentation. The culture medium comprises maltose, yeast extract powder, Ca<2+> and vitamin E. The method comprises that the culture medium is inoculated with strain fermentation liquor, 50ml of the culture medium loaded in a triangular flask having the capacity of 250ml is used, and an inoculation amount ratio is 5%; and the strain fermentation liquor is cultured at a temperature of 25 DEG C at a rotation rate of 140r / min. The method comprises strain activation, strain fermentation liquor culture and fermentation optimization. The culture medium and the method can greatly improve an amylase yield.

Description

technical field [0001] The invention relates to a culture medium and a method for high-yielding amylase fermented by Glomus melii. Background technique [0002] Amylase (amylase) is a general term for hydrolyzing starch and glycogen enzymes. It is a class of enzymes that can hydrolyze starch and glycogen molecules into dextrin and smaller molecules. It is widely found in animals, plants and microorganisms. Amylase is widely used in industries such as fuel ethanol, starch syrup, traditional brewing, food processing, and textile desizing. [0003] Although amylase exists in both animals and plants, it is difficult to industrialize due to the complicated extraction process and high cost. The extraction process of amylase derived from microorganisms is simple, low in cost, high in yield, [0004] Due to its short cycle time, stable properties, and mild operating conditions, amylase derived from microorganisms has been widely used in industry, especially in the starch hydrolysi...

Claims

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Application Information

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IPC IPC(8): C12N9/30C12R1/645
CPCC12N9/242C12Y302/01001
Inventor 邱君志李小霞曹丽萍孟丽雪郭庆丰何肖云张以盼涂洁董冬姚灵丹
Owner FUJIAN AGRI & FORESTRY UNIV
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