Rumen clostridium using lactic acid for producing hexanoic acid and application thereof

A rumen bacterium, a technology for use, applied in the field of microorganisms, to achieve good application prospects, good growth stability, and the effect of improving flavor and quality

Active Publication Date: 2016-03-23
SICHUAN JIANNANCHUN GRP CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, current research shows that when most microorganisms use lactic acid as a substrate, they can only produce butyric acid or propionic acid, and basically do not produce caproic acid or other medium-chain fatty acids.

Method used

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  • Rumen clostridium using lactic acid for producing hexanoic acid and application thereof
  • Rumen clostridium using lactic acid for producing hexanoic acid and application thereof
  • Rumen clostridium using lactic acid for producing hexanoic acid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Isolation, purification and cultivation of embodiment 1 bacterial classification

[0028] 1. Culture medium for isolation and culture

[0029] Medium composition (per 100mL): 1g casein peptone, 0.25g yeast juice, 0.4g NaHCO 3 ,0.1g cysteine,0.045gK 2 HPO4, 0.045gKH 2 PO 4 ,0.09gNaCl,0.009gMgSO 4 ·7H 2 O,0.009gCaCl 2 , 0.1mg resazurin, 1mg oxyhemoglobin, 0.5mL lactic acid, 0.1ml trace element solution, 1ml vitamin solution. Distilled water 100ml. in:

[0030] Trace element solution (per L): FeCl 2 .4H 2 O1.5g; CoCl 2 .6H 2 O0.19g; MnCl 2 .4H 2 O0.1g; ZnCl 2 0.07g; NaMO 4 .2H 2 O0.036g; NiCl 2 .6H 2 O0.024g; H 3 BO 3 0.006g; CuCl 2 .2H 2 O0.002g; HCl (25% solution) 10ml.

[0031] Vitamin solution (per L): p-aminobenzoic acid 0.04g; biotin 0.01g; niacin 0.1g; B 5 0.05g; B 6 0.15g; B 1 0.1g; B 12 0.05g; D.L-lipoic acid 0.03g; B 2 0.03g; folic acid 0.01g.

[0032] 2. Separation process

[0033] (1) Enrichment of strains

[0034] Take 1g of pit...

Embodiment 2

[0043] The cultivation and identification of embodiment two strains

[0044] 1. Morphological characteristics

[0045] Colony morphology: Clostridium CGMCCNo:9926 was cultivated on the plate for 7 days, the colony was round, milky white, smooth and moist, slightly raised, sticky, easy to pick, and the diameter of the colony was 1.5mm.

[0046] Bacteria morphology: Gram staining is positive, the bacteria are spherical, with a diameter of 2.0-2.5um. SEM photos see figure 1 (SEM, 5200×)

[0047] 2. Physiological and biochemical characteristics

[0048] with BiologAN MicroPlates TM The physiological and biochemical characteristics of the Clostridium are shown in Table 1 through analysis with the API20A anaerobic identification system.

[0049] Table 1 Physiological and biochemical characteristics of CPC-11

[0050]

[0051]

[0052] 3. Molecular biological characteristics

[0053] The bacterial 16SrRNA universal primer pair 27f-1492r was used to amplify the 16SrRNA of...

Embodiment 3

[0060] Example 3 The impact test of the initial pH value on the production of caproic acid

[0061] Adopt liquid culture medium (see embodiment 1), add lactic acid to final concentration and be 10g / L, regulate the pH value of culture medium with sodium hydroxide and hydrochloric acid, obtain initial pH and be respectively 5.5, 6.0, 6.5, 7.0, 7.5, 8.0 altogether 6 acid-base gradients, filled with N 2 15 minutes, then dispense 50mL into 250ml anaerobic bottles, and sterilize at 121°C for 15min. Inoculate CPC-11 bacterial solution by 5% (v / v) (concentration is about 10 8 CFU / mL), cultured in a shaker at 30° C. at 150 rpm for 7 days, and measured the hexanoic acid production and conversion rate. The results are shown in Table 3.

[0062] Table 3, the influence of different pH on producing caproic acid

[0063]

[0064]

[0065] It can be seen from the table that CPC-11 can use lactic acid to produce caproic acid well in the pH range of 5.0-6.5, and the optimum pH for pro...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a new rumen clostridium CPC-11(Clostridium sp.CPC-11) and application thereof. The new rumen clostridium Clostridium sp.CPC-11 is capable of converting lactic acid into hexanoic acid, using lactic acid or yellow water for producing hexanoic acid through fermentation, and being used in intensified culture of artificial pit mud or the process of increasing hexanoic acid and reducing lactic acid of pit fermentation, the yield of hexanoic acid is increased, and the rumen clostridium has excellent application prospects.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a novel Clostridium sp. CPC-11 and the application of the bacteria in the production of hexanoic acid. Background technique [0002] Caproic acid (n-caproate) is a medium-chain fatty acid containing six carbon atoms (C 5 h 11 COOH), with strong hydrophobicity and high energy density, is an important chemical raw material, which can be used in the fields of food spices, animal feed, green antibacterial agents and corrosion inhibitors. In particular, caproic acid is an important precursor for the synthesis of ethyl caproate (the main aroma substance of Luzhou-flavor liquor quality). Its ester products are also widely used in resin, rubber, pharmaceutical and other industries. [0003] Microbial catalyzed production of caproic acid is a carbon chain elongation process. Clostridium klyuveri is the most studied and most in-depth one. This bacterium uses ethanol as the elec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/40C12G3/02C02F3/34C12R1/145C02F103/32
CPCC02F3/34C02F2103/32C12G3/02C12P7/40C12N1/205C12R2001/145
Inventor 徐姿静陶勇徐占成何晓红唐清兰樊科权
Owner SICHUAN JIANNANCHUN GRP CO LTD
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