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Bacterial strain for preparing 3-hydroxy butanone and uses thereof

The technology of hydroxybutanone and strain is applied in the field of 3-hydroxybutanone-producing strains, which can solve problems such as difficulty in producing 3-hydroxybutanone by fermentation, and achieve the effect of alleviating the shortage of petrochemical resources and being environmentally friendly.

Inactive Publication Date: 2008-10-29
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This has directly led to the difficulty of utilizing these microbial strains for industrialized fermentation to produce 3-hydroxybutanone

Method used

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  • Bacterial strain for preparing 3-hydroxy butanone and uses thereof

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Experimental program
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Effect test

Embodiment 1

[0041] Collect soil samples from the surface of the orchard in Shuaiqi Farm, Pukou District, Nanjing, weigh 3.0 grams, bake at 90°C for 36 hours, add to 100 mL of sterile water to make a suspension; take 0.5 mL of the bacterial suspension and transfer it to a 150 mL Erlenmeyer flask, The volume of the enrichment medium in the Erlenmeyer flask was 50mL. After the enrichment culture was carried out at 35°C for 24h, the culture medium was taken and diluted to 10 -6 , take 0.1mL of the diluted solution and spread it on a plate containing enriched medium, and after culturing at 35°C for 24 hours, pick a single colony and conduct V-P (Vagex-Proskauer) experiments one by one. The specific method is: inoculate the strain to be tested on In a 10mL culture tube filled with 5mL of V-P experimental medium, culture at 35°C for 4 days, take 2.5mL of culture medium, first add 0.6mL of pure alcohol solution containing 5% α-naphthol, then add 40% potassium hydroxide Aqueous solution 0.2mL, sha...

Embodiment 2

[0046] The 77 strains of V-P positive strains obtained in Example 1 were transferred one by one to a screening plate equipped with a separation medium with a sterilized toothpick, cultivated in an incubator at a temperature of 37°C, and after cultivating for 24 hours, it was found that there were The 28 corresponding strains can grow normally, and these 28 single colonies were collected respectively, numbered as: ME-J1~ME-J28, and cultured in 10mL culture tubes containing 5mL fermentation medium for 24h; after the culture, 12000r / Centrifuge for 5 minutes, and measure the content of 3-hydroxybutanone in the supernatant by HPLC. It is found that most of the strains produce more lactic acid and acetic acid, and a few strains produce more 3-hydroxybutanone. Among them, the strain ME-J25 produces 3-hydroxybutanone. Butanone is most notable.

[0047] Metabolite distribution of table 1V-P positive strains

[0048]

[0049] Note: ND means "not detected".

[0050] in:

[0051] S...

Embodiment 3

[0054] To the ME-J25 bacterial strain obtained by screening in Example 2, press "Common Bacterial System Identification Manual" (Dong Xiuzhu, Cai Miaoying, etc. Common Bacterial System Identification Manual (First Edition). Beijing: Science Press, 2001, p 59) Physiological and biochemical characteristics were identified (Table 2). According to the comparison of morphology and physiological and biochemical characteristics in Table 2, the strain ME-J25 was identified as Paenibacillus polymyxa, which has been preserved in the China Microbiological Culture Collection Management Committee General Microbiology Center, the preservation number is CGMCC No.2505

[0055] Table 2 Comparison table of identification results of physiological and biochemical characteristics of strain ME-J25

[0056]

[0057] Note: +: >90% strains are positive; -: >90% strains are negative; ++: produce acid and gas

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Abstract

The invention discloses a strain for generating 3-hydroxy butanone and the application thereof, which belong to the field of bioengineering technology. The stain is named as Paenibacillus polymyxa ME-J25 and is preserved in China General Microbiological Culture Collection Center with the preservation number of CGMCC No.2505. The strain can be fermented to generate 3-hydroxy butanone with higher concentration, higher transformation efficiency and extremely low content of the reduced byproduct 2,3-butanediol of 3-hydroxy butanone, and is suitable for industrial production of 3-hydroxy butanone.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a bacterial strain producing 3-hydroxybutanone and an application thereof. Background technique [0002] 3-Hydroxybutanone (3-Hydroxybutanone), also known as acetoin (Acetoin), methyl acetylmethanol (Acetylmethylcarbinol), naturally exists in dairy products and some fruits, is a widely used edible flavor, my country GB2760- 86 stipulates that it is allowed to be eaten, and the safety number of the American Food and Extraction Association (FEMA) is 2008 (Ling Guanting. Food Additive Handbook (Second Edition) [M]. Beijing: Chemical Industry Press, 1997, 301-301). 3-Hydroxybutanone has a wide range of applications as a spice, and the dosage is also relatively large, so it is favored by perfumers. Its main application is for the production of flavors such as cream, dairy products, yogurt and strawberry type (Hu Mingyi , Wang Zhong. Fine and Specialty Chemicals, 2002, 10(1): 20-2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/26C12R1/12
Inventor 黄和纪晓俊胡南李霜朱建国杜军任潇
Owner NANJING TECH UNIV
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