A kind of Aureobasidium pullulans using starch to produce polymalic acid and its application

A technology of Aureobasidium pullulans and polymalic acid, applied in the direction of microorganism-based methods, microorganisms, microorganisms, etc., can solve the problems of difficult quality control, high cost of raw materials, and increased operating costs of fermentation equipment, so as to reduce the consumption of cooling water , Strain nutritional requirements are simple, the effect of good industrial production value

Active Publication Date: 2020-12-08
GUANGXI UNIV
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Problems solved by technology

[0004] It can be seen that many researchers have obtained many strains of Aureobasidium pullulans capable of synthesizing polymalic acid through natural breeding or mutation breeding, but through comparison, it is found that these strains still have some shortcomings in the process of fermenting and producing polymalic acid: (1) High-concentration glucose is used as a carbon source. On the one hand, the cost of raw materials is high, and on the other hand, high-concentration glucose has a substrate inhibitory effect on the growth of bacteria, which limits the increase in the production of polymalic acid; when molasses is used as a carbon source, due to Molasses itself has a darker color and more impurities. Although the cost of raw materials has decreased, it will increase the cost of extraction and purification of polymalic acid and treatment of fermentation waste liquid
(2) Although the yield of polymalic acid is as high as 62.27g / L, sodium fumarate needs to be added as a fermentation accelerator in the medium, which increases the cost of fermentation raw materials
(3) A certain amount of corn steep liquor or corn extract must be added to the fermentation medium of the above three types of Aureobasidium pullulans. On the one hand, the cost of fermentation raw materials is increased. On the other hand, due to the complex composition of corn steep liquor or corn extract, the quality is difficult to control , there is uncertainty about the effect on the fermentation process
(4) The fermentation temperature of the above three types of Aureobasidium pullulans is 25°C, and microbial fermentation is an exothermic process, so a large amount of cooling water is required to maintain the lower fermentation temperature, thereby increasing the operating cost of fermentation equipment
Although Aureobasidium pullulans ( Aureobasidium pullulans ) GXZ-6 can use tapioca starch, corn starch, wheat starch, soluble starch, starch saccharification solution, and maltose syrup as carbon sources to ferment polymalic acid, and it can be produced in batches at 30 °C More than 60g / L polymalic acid can be obtained by fermentation, but it produces melanin during the fermentation process, which usually makes the downstream separation and extraction of metabolites difficult, affecting the yield of polymalic acid product and product appearance

Method used

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  • A kind of Aureobasidium pullulans using starch to produce polymalic acid and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Aureobasidium pullulans ( Aureobasidium pullulans ) GXL-1 utilizes tapioca starch to produce polymalic acid, including the following steps:

[0030] (1) Strain activation: Aureobasidium pullulans preserved in glycerol ( Aureobasidium pullulans ) GXL-1 was streaked on PDA slant medium and incubated at 30°C for 2 days.

[0031] (2) Seed cultivation: Scrape a ring on the slanted surface of the activated strain, inoculate it into a 500mL shaker flask containing 80mL of liquid seed medium, and cultivate for 36h at a shaker speed of 220rpm and a temperature of 25ºC to obtain seed solution. Described liquid seed medium is glucose 100g / L, ammonium chloride 2g / L, KCl 0.6g / L, KH 2 PO 4 0.2g / L, MgSO 4 ·7H 2 O 0.3g / L, ZnSO 4 ·7H 2 O 0.2g / L, CaCO 3 30g / L.

[0032] (3) Fermentation culture: The seed liquid obtained from the culture was placed in a 250mL conical flask containing 30mL liquid fermentation medium according to the inoculation amount of 8% (v / v), and cultured f...

Embodiment 2

[0035] Aureobasidium pullulans ( Aureobasidium pullulans ) GXL-1 utilizes wheat starch to produce polymalic acid, including the following steps:

[0036] (1) Strain activation: Aureobasidium pullulans preserved in glycerol ( Aureobasidium pullulans ) GXL-1 was streaked on PDA slant medium and incubated at 30°C for 2 days.

[0037] (2) Seed cultivation: Scrape a ring on the slanted surface of the activated strain, inoculate it into a 500mL shaker flask containing 80mL of liquid seed medium, and cultivate for 36h at a shaker speed of 220rpm and a temperature of 25ºC to obtain seed solution. Described liquid seed medium is glucose 100g / L, ammonium chloride 2g / L, KCl 0.6g / L, KH 2 PO 4 0.2g / L, MgSO 4 ·7H 2 O 0.3g / L, ZnSO 4 ·7H 2 O 0.2g / L, CaCO 3 30g / L.

[0038] (3) Fermentation culture: The seed liquid obtained from the culture was placed in a 250mL conical flask containing 30mL liquid fermentation medium according to the inoculation amount of 8% (v / v), and cultured for...

Embodiment 3

[0041] Aureobasidium pullulans ( Aureobasidium pullulans ) GXL-1 utilizes cornstarch to produce polymalic acid, which involves the following steps:

[0042] (1) Strain activation: Aureobasidium pullulans preserved in glycerol ( Aureobasidium pullulans ) GXL-1 was streaked on PDA slant medium and incubated at 30°C for 2 days.

[0043] (2) Seed cultivation: Scrape a ring on the slanted surface of the activated strain, inoculate it into a 500mL shaker flask containing 80mL of liquid seed medium, and cultivate for 36h at a shaker speed of 220rpm and a temperature of 25ºC to obtain seed solution. Described liquid seed medium is glucose 100g / L, ammonium chloride 2g / L, KCl 0.6g / L, KH 2 PO 4 0.2g / L, MgSO 4 ·7H 2 O 0.3g / L, ZnSO 4 ·7H 2 O 0.2g / L, CaCO 3 30g / L.

[0044] (3) Fermentation culture: The seed liquid obtained from the culture was placed in a 250mL conical flask containing 30mL liquid fermentation medium according to the inoculation amount of 8% (v / v), and cultured ...

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Abstract

The invention discloses a kind of Aureobasidium pullulans that utilizes starch to produce polymalic acid and its application. Aureobasidium pullulans ) GXL‑1, the deposit number is CCTCC NO: M 2018519, the deposit date is August 1, 2018, the deposit unit: China Center for Type Culture Collection, deposit address: China. Wuhan. Wuhan University. The Aureobasidium pullulans of the present invention ( Aureobasidium pullulans ) GXL‑1 can directly use raw starch or starch liquefaction liquid as the carbon source. Without adding fermentation promoters and growth factors, batch fermentation at 30°C can obtain up to 57g / L of polymalic acid, and no fermentation process occurs. The production of melanin has the advantages of low fermentation cost, easy product purification and simple process, and has great potential for industrial production.

Description

technical field [0001] The invention belongs to the technical field of microorganism application, in particular to Aureobasidium pullulans for producing polymalic acid by utilizing starch and application thereof. Background technique [0002] Polymalic acid [poly(β-L-malic acid), PMA] is a new type of polymer compound obtained by dehydrating polyester with α-hydroxyl and β-carboxyl groups with L-malic acid as the only monomer. It has good properties. Water solubility, biodegradability, biocompatibility and chemical modification have broad application prospects in medicine, food, agriculture and environmental management. The production methods of polymalic acid include chemical synthesis and biosynthesis. The chemical synthesis of polymalic acid has complex steps, many by-products, small molecular weight and high cost; while the biological synthesis of polymalic acid has simple operation, fewer by-products, large product molecular weight and low cost, so the use of biologica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12P7/62C12R1/645
CPCC12P7/625C12N1/145C12R2001/645
Inventor 曾伟梁智群陈桂光张斌蒋莉刘瑶李凌甫
Owner GUANGXI UNIV
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