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Acetobacter pasteurianus with excellent heat resistance and application thereof

A technology of Acetobacter pasteuriani and its ability, applied in the direction of bacteria, vinegar preparation, and microbial-based methods, can solve the problems of unreported high-temperature resistance, too expensive fruit vinegar, and low acetic acid production, so as to achieve plumpness, The color is orange-yellow and clear, with a wide tolerance range

Active Publication Date: 2016-11-30
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] 30°C, although acetic acid bacteria can also grow at 37-40°C (Gullo et al.2008), but the growth is slow
In summer, the temperature is high, and the commonly used acetic acid bacteria AS1.41 and Huniang 1.01 are difficult to withstand high temperature, and the output of acetic acid is not high. The production of the above fruit vinegars requires high cooling costs. too expensive
In order to improve the level of acetic acid fermentation under high-temperature conditions, the improvement of the heat-resistant function of acetic acid bacteria has become a research hotspot, such as the Chinese patent application number CN201310549668, "High-temperature-resistant acetic acid bacteria and its application in the production of acetic acid by fermentation", in which the acetic acid Bacteria can grow at 45°C (high temperature resistance)), the ethanol-acetic acid conversion rate at 37°C and 7% ethanol is 88.8%, and the ethanol-acetic acid conversion rate at 37°C and cider (7% ethanol) is 93% %, there is no report of acetic acid bacteria with superior high temperature resistance

Method used

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  • Acetobacter pasteurianus with excellent heat resistance and application thereof
  • Acetobacter pasteurianus with excellent heat resistance and application thereof
  • Acetobacter pasteurianus with excellent heat resistance and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Preliminary separation and purification of high-temperature-resistant high-alcohol acetic acid bacteria

[0020] In order to clearly illustrate the technical features of the solution of the present invention, the present invention will be described below in conjunction with specific examples. However, the protection scope of the present invention is not limited to these examples. All changes or equivalent substitutions that do not depart from the concept of the present invention are included in the protection scope of the present invention.

[0021] Selection of sources of acetic acid bacteria isolation:

[0022] (1) Vinegar grains are from Danyang Vinegar Factory;

[0023] (2) Preliminary separation and purification of high-temperature-resistant high-alcohol acetic acid bacteria

[0024] ① Enrichment culture of acetic acid bacteria:

[0025] Add 1g of vinegar unstrained spirits from a vinegar factory (multiple) with traditional solid-state fermentation te...

Embodiment 2

[0032] Example 2 Identification of acetic acid bacteria DY4

[0033] (1) According to the "Common Bacteria System Identification Manual" and "Bergey's Bacterial Identification Manual" eighth edition, the strains were identified morphologically, physiologically and biochemically (Table 1).

[0034] Table 1 Physiological and biochemical characteristics of DY4 and AS1.41

[0035]

[0036] (2) Identification of 16Sr DNA of acetic acid bacteria

[0037] Pick acetic acid bacteria and inoculate them in 10mL / 100mL Erlenmeyer flask liquid culture medium, shake and culture in a constant temperature shaking incubator at 30°C and 180r / min for 24h. Take 2mL of the bacterial solution in a centrifuge tube, centrifuge at 12000r / min for 2min, remove the supernatant, and extract the genome of the strain according to the instructions of the SK1201-MNIQ-10 Column Bacterial Genome DNA Extraction Kit from Shanghai Sangon Bioengineering Technology Service Co., Ltd. DNA, as a template. Select u...

Embodiment 3

[0046] Embodiment 3 DY4 acetic acid bacteria high temperature resistance experiment

[0047] Take the logarithmic period Acetobacter pasteμ rianμ s DY4 、 Add 4mL of AS1.41 bacteria solution to 96mL liquid medium (2% glucose, 2% yeast powder, 0.01% MgSO 4 , 4% absolute ethanol) in a 500mL Erlenmeyer flask, respectively at 30°C, 33°C, 35°C, 37°C, 40°C, 41°C, 42°C, 43°C, 44°C in a small shaking culture, shaker The speed is 180r / min, and the growth amount OD of the bacteria is measured every day 600 value and acid production until growth OD 600 Until the value and acid production reach a stable level, draw a graph, compare the final growth and acid production, and obtain the limit temperature tolerated by the strain ( Figure 4 , Figure 5 ).

[0048] Conclusion: Bacteria DY4 can have relatively stable growth and acid production (ethanol-acetic acid conversion rate>90.6%) at 30-40°C, the optimum fermentation temperature is 37°C, and the maximum conversion rate can be reac...

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Abstract

The invention discloses acetobacter pasteurianus DY4 with excellent heat resistance and application thereof. The strain was preserved in China Center for Type Culture Collection (Wuhan University, Wuhan, China), and the preservation number is CCTCC NO: M 2015462. Under the condition of 4% ethanol, the acetobacter pasteurianus can have stable growth and acid yield at 30-40 DEG C (ethanol-acetic acid conversion rate is greater than 90.6%), the maximum conversion rate reaches 97.5% at 37 DEG C, OD600 can reach 0.72 at 43 DEG C, acid yield can reach 16.5 g / L (the maximum conversion rate is 40.0%); and at 37 DEG C and under the condition of 8% ethanol, ethanol conversion rate reaches 95.6%. The strain of the invention is suitable for brewing of fruit vinegar in summer, and a vinegar product has superior flavor.

Description

technical field [0001] The invention belongs to the field of microorganism and fermentation industry, and in particular relates to a kind of Acetobacter pasteurii with excellent heat resistance Acetobacer pasteurianus DY4 and its uses. Background technique [0002] Acetic acid bacteria (AAB) are obligate aerobic Gram-negative bacteria capable of oxidizing acetic acid from ethanol (Deppenmeier et al. 2010). AAB is commonly isolated from natural habitats and from vinegar fermented grains (Wu et al. 2010). During the fermentation process, acetic acid bacteria will be subjected to various stresses, such as high temperature. The optimum growth temperature for AAB is 25- [0003] 30°C, although acetic acid bacteria can also grow at 37-40°C (Gullo et al. 2008), but the growth is slow. The heat of fermentation accumulated during the fermentation of acetic acid bacteria will inevitably make the fermentation temperature higher than 37 °C, resulting in slow cell growth and low ace...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12J1/04C12R1/02
CPCC12J1/04C12N1/205C12R2001/02
Inventor 胡勇徐宁李冬生周梦舟祁勇刚高冰汪超石勇徐梅朱于鹏龚元元
Owner HUBEI UNIV OF TECH