Method for producing butyric acid through fermentation

A technology of butyric acid and Clostridium tyrobutyricum is applied in the directions of microorganism-based methods, fermentation, biochemical equipment and methods, etc., and can solve the problems of reducing the yield of butyric acid, high cost, and low concentration of fermentation products.

Inactive Publication Date: 2019-02-19
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the substrates of butyric acid fermentation are mainly glucose and agricultural waste (such as bagasse and corn husk) hydrolyzate. Although the latter solves the problem of raw material sources, due to the large amount of toxic inhibitors produced in the pretreatment process, the fermentation products low concentration
While detoxification can mitigate this effect, it results in loss of sugar and additional operating costs
There is another bottleneck in the fermentation of the above two substrates: the existence of the by-product acetic acid not only reduces the yield of butyric acid, but also makes the subsequent separation and purification steps cumbersome and costly

Method used

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  • Method for producing butyric acid through fermentation
  • Method for producing butyric acid through fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Effects of Different Proportions of Glucose and Mannitol Mixed Substrate Fermentation on Butyric Acid Fermentation

[0035] (1) Preparation of culture medium

[0036] CGM (Clostridium Growth Medium) medium (1L system): 2g yeast extract, 4g tryptone, 2g (NH 4 ) 2 SO 4 , 1g K 2 HPO 4 , 0.5g KH 2 PO 4 , 0.1g MgSO4 ·7H 2 O, the carbon source is glucose and / or mannitol, and the carbon and nitrogen are sterilized separately from the medium (121°C, 20min).

[0037] (2) Shake flask fermentation experiment

[0038] In a 100mL anaerobic bottle (filled with 50mL sterile CGM medium without carbon source and 2g / L CaCO 3 ) in different proportions, sterilized glucose and / or mannitol were added as carbon sources. Clostridium tyrobutyricum ATCC 25755 (American Type Culture Collection, American Type Culture Collection) was activated and cultured in CGM medium overnight, and then the seed solution was inserted into the anaerobic bottle, then cultured at 37 °С, 150 r...

Embodiment 2

[0041] Embodiment 2: the preparation of kelp hydrolyzate

[0042] (1) Determination of kelp components

[0043] Kelp was purchased from the Vegetable Market in Suishi Village, University City, Guangzhou. The determination of moisture, total ash, protein, fat and total sugar content in kelp was determined according to GB5009.3-2016, GB5009.4-2016, GB5009.52010, GB5009.6-2016 respectively and phenol-sulfuric acid method. The specific ingredients are shown in the table below:

[0044]

moisture

total ash

protein

Fat

total sugar

content(%)

9.56

28.67

7.21

1.30

58.95

[0045] (2) Preparation of kelp acid hydrolyzate

[0046] The dry kelp was crushed into powder by mechanical pulverization, passed through a 100-mesh sieve and stored in an oven at 65°C for drying, and then the above-mentioned kelp powder was mixed with 0.1M H 2 SO 4 well mixed. Pretreatment at 121°C for 45 minutes, after cooling, the hydrolyzed solution was...

Embodiment 3

[0049] Example 3: Clostridium tyrobutyricum uses glucose, glucose / mannitol mixture, and kelp enzymatic hydrolyzate as substrates to ferment and produce butyric acid respectively

[0050] Glucose (~90g / L), glucose / mannitol mixed substrate (~60g / L glucose and ~30g / L mannitol), kelp enzymatic hydrolysis solution (ammonium acetate in the medium instead of ammonium sulfate, and glucose , make glucose:mannitol=44:23) as the carbon source of the fermentation experiment. The fermentation experiment was carried out in a 5L fully automatic mechanical stirring fermenter. Before fermentation, first sterilize the fermenter device at 121°C for 20 minutes, then add CGM medium (1L), and sterilize it at 121°C for 20 minutes, and after cooling, replace the air in the tank with sterile nitrogen to achieve anaerobic environment. Clostridium tyrobutyricum ATCC25755 was activated and cultured overnight in CGM medium, then the seed liquid was introduced into the fermenter according to the inoculum...

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Abstract

The invention belongs to the technical field of biology, and discloses a method for producing butyric acid through fermentation. The method comprises the following steps that 1, Clostrdium tyrobutyricum is activated, and a seed solution is prepared; 2, a mannitol and glucose mixture is adopted as a carbon source, a liquid culture medium is prepared, and sterilization is performed; 3, the seed solution in step 1 is inoculated in the liquid culture medium in step 2, and cell fermentation is performed on the conditions that the temperature is 35-39 DEG C, pH is 5-7 and the rotating speed is 100-300 rpm to obtain butyric acid. The accumulation of by-product butyric acid can be lowered, the conversion rate and the selectivity of the target product butyric acid are improved, and therefore the follow-up separation and purification cost can be lowered. In addition, the mixed substrate can be replaced with brown alga hydrolysate or brown alga processing waste, and therefore efficient biologicalconversion from cheap marine biomass to high-selectivity butyric acid is achieved.

Description

technical field [0001] The invention relates to a method for using Clostridium tyrobutyricum to ferment and produce highly selective butyric acid with glucose and mannitol as a mixed substrate, belonging to the technical field of organic acid fermentation. Background technique [0002] Butyric acid, also known as butyric acid, refers to a straight-chain saturated monobasic fatty acid containing four carbon atoms, which often exists in nature in the form of free or ester. Butyric acid and its derivatives are widely used in chemical, food, pharmaceutical and feed industries. [0003] At present, the production of butyric acid mainly has two methods: chemical synthesis and microbial fermentation. In the industrial production of butyric acid, propylene oxo synthesis and n-butyraldehyde oxidation are the most commonly used methods. Among them, the propylene carbonyl synthesis method mainly converts propylene into butyraldehyde through glycosylation, and then oxidizes butyraldeh...

Claims

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Application Information

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IPC IPC(8): C12P7/52C12R1/145
CPCC12P7/52
Inventor 王菊芳傅宏鑫廖正平索玉凯张亚南胡佳磊郭晓龙
Owner SOUTH CHINA UNIV OF TECH
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