Expansion penicillium strain as well as culture method and use thereof

A technology for Penicillium dilatans and strains, which is applied to Penicillium dilatans strain and its cultivation method and application field in the production of galactooligosaccharides, and can solve the problems of few batches used, poor stability and the like

Active Publication Date: 2010-06-09
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are differences in the stability of immobilized β-galactosidase from different sources. For example, the stability of immobilized enzymes from some bacteria is poor, and fewer batches are used.

Method used

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  • Expansion penicillium strain as well as culture method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Penicillium dilatantum F3 strain, the preservation number of which is CGMCC No.2352 in the General Microbiology Center of China Committee for Culture Collection of Microorganisms.

[0036] (1) Observation of morphological characteristics

[0037] The spores of Penicillium expansica F3CGMCC No.2352 were picked with an inoculation loop and placed on the potato medium (PDA) plate and the Chase medium plate respectively, and cultured at 30°C. On the PDA plate, a large number of green spore groups are produced in 2 to 3 days, and the colony spreads; on the Chase plate, the colony grows rapidly, 5 to 6 cm in diameter, 1 to 2 mm thick, and radial grooves in 12 to 14 days , white edge, many spores, olive green, light pink in late growth period, less exudate, no special smell, bright yellow to reddish brown on the reverse side. The conidiophores are smooth, the bristle-like branches are asymmetrical, the conidia are born in chains on the small stalks, and the conidia are ellips...

Embodiment 2

[0044] A kind of culture method of Penicillium dilatantum F3 bacterial strain, concrete steps are as follows:

[0045] Take Penicillium expansica F3 (strain number CGMCC No.2352) and inoculate it into a fresh slant culture medium for activation for 48 hours at a temperature of 28°C. Take the activated Penicillium expanse F3 and inoculate it in 5 mL of seed culture solution. After culturing for 30 hours, Transfer to 500mL enzyme-producing fermentation broth and expand culture for 40 hours at a culture temperature of 28°C and a shaker speed of 180 rpm. Penicillium expanses F3CGMCC No.2352 grows and produces enzymes well in the above medium.

[0046] The composition of the slant medium: glucose 5g / L, peptone 5g / L, yeast powder 5g / L, agar 20g / L, pH natural, sterilized at 115°C for 30 minutes.

[0047] The composition of the seed culture solution: glucose 5g / L, peptone 5g / L, yeast powder 5g / L, pH natural, sterilized at 115°C for 30 minutes.

[0048] The composition of the enzyme-...

Embodiment 3

[0051] A method for preparing immobilized enzymes to produce galacto-oligosaccharides by Penicillium expanses F3 strain, the specific steps are as follows:

[0052] The method described in Example 2 was used to cultivate Penicillium dilatantum F3 (bacteria preservation number CGMCC No.2352), and the bacterial cells were obtained by filtration, and the bacterial cells were suspended in pH6.4, 50mM pH6.4, 50mM Phosphate buffer solution, after being completely frozen below -20°C, thawed at room temperature, and then repeated freezing and thawing twice, the resulting cell suspension was used as the source of β-galactosidase enzyme;

[0053] Take the frozen-thawed cell suspension and immobilize it with calcium alginate embedding method, mix the bacterial suspension with 3% (w / w) sodium alginate solution at a ratio of w:v=1:2, drop Put into 3% (w / w) calcium chloride solution and harden at 4°C to prepare the immobilized enzyme.

[0054] The immobilized enzyme was mixed with 380g / L l...

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Abstract

The invention discloses a strain of penicillium expansum F3 and the application of the penicillium expansum F3 in preparing oligo-galactose. The preservation number of the strain is CGMCC No. 2352, abacterial colony thereof grows rapidly with radiation rills and olive green color, the strain is light pink in the late growth stage, a conidiophore is smooth, broom-shaped branches are not symmetrical, a conidia is burn on a little peduncle with a chain shape, and the conidia takes an ellipsoid shape. Immobilized enzyme which is obtained through taking freeze-thaw whole cells of the strain as anenzyme source and adopting an alginate calcium embedding method is very stable and can be repeatedly used to synthesize the oligo-galactose, and the immobilized enzyme reacts for 11 batches and keepsthe output more than 20%. The strain of the invention can be simply cultured, the hereditary property is stable, the method for preparing the immobilized enzyme is convenient, the property of enzyme is stable, the advantages for applying method in industrial production of the oligo-galactose are prominent, and the prospect is optimistic.

Description

technical field [0001] The invention relates to a Penicillium strain, a culture method and application thereof, in particular to a Penicillium dilatantum strain, a culture method thereof and an application in the production of galacto-oligosaccharides. technical background [0002] Galacto-oligosaccharides (GOS) are a kind of non-digestible oligosaccharides, which have many beneficial physiological functions. There are outstanding advantages in terms of less gas and so on. This type of oligosaccharide can be synthesized by β-galactosidase produced by microorganisms using lactose as a substrate to transglycosylate. There are two preparation methods, one is using free enzymes, and the other is using immobilized enzymes. The free enzyme is unstable during the reaction process, and it is difficult to reuse, so it is not suitable for industrial production, while the immobilized enzyme can effectively improve the stability of the enzyme, can be reused, and is suitable for industr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N11/04C12P19/04C12R1/80
Inventor 肖敏卢丽丽李玉梅李正义
Owner SHANDONG UNIV
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