Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Providencia for synthesizing ethanol-resistant urethanase and urease

A technology of urethane and Providencia bacteria, applied in the field of Providencia bacteria, can solve the problems such as difficult to completely eradicate EC, and achieve the effect of huge application potential and high-efficiency enzyme activity

Inactive Publication Date: 2016-09-28
JIANGNAN UNIV
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since urea is not the only precursor substance that forms EC, it is difficult to completely eradicate the formation of EC by this method.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Providencia for synthesizing ethanol-resistant urethanase and urease
  • Providencia for synthesizing ethanol-resistant urethanase and urease
  • Providencia for synthesizing ethanol-resistant urethanase and urease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Providencia screening method

[0033] Take 5g of the soil sampled around the chemical plant, dissolve it in 10ml of 0.9% physiological saline, and break it up with glass beads. Take 1ml of the supernatant and inoculate it into 25ml enrichment medium (beef extract 10g / L, peptone 10g / L, sodium chloride 15g / L, ethyl carbamate 10g / L) at a temperature of 30°C and a rotation speed of 200r. min -1 Incubate for 24 hours on a shaker. The above culture solution was transferred to fresh enriched medium with 1% inoculum amount and continued to culture for 24 hours. The enriched bacterial solution was serially diluted (10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7) on a screening plate (beef extract 10g / L, peptone 10g / L, sodium chloride 15g / L, ethyl carbamate 30g / L, agar 20g / L), and cultivate at 30°C until obvious colonies grow . Pick colonies with larger colonies and transfer them to fresh nutrient broth medium (peptone 10g / L, beef extract 3g / L, NaC...

Embodiment 2

[0035] Embodiment 2: the preparation of crude enzyme liquid

[0036] Pick the strains stored at -80°C and streak on the nutrient broth solid medium, culture at 30°C for 20 hours, take a single colony and inoculate it in fresh nutrient broth medium at 30°C with a rotation speed of 200r min- 1 Continue culturing for 20 h on a shaker. Centrifuge the bacterial solution at 10000rpm at 4°C for 5min, discard the supernatant, wash the bacterial cells and resuspend them in 50mM citric acid buffer solution with pH 4.5, the final concentration of the bacterial cells is 0.1g / ml. Place the cells in an ice-water bath for ultrasonic crushing. The conditions for ultrasonic crushing are: power 70W, work for 2 seconds with an interval of 4 seconds, and repeat the work for 30 minutes until the cell solution becomes clear. Centrifuge the crushed solution at 12000rpm at 4°C for 10min, transfer the supernatant to a clean centrifuge tube and store it at low temperature.

Embodiment 3

[0037] Embodiment 3: the mensuration of crude enzyme liquid urethane hydrolase and urease activity

[0038] The crude enzyme solution obtained in Example 2 was tested for urethane hydrolase and urease enzyme activity respectively according to the enzyme solution assay method described in Materials and Methods. The result is as figure 1 As shown, the urease activity is 1.62U / mL, and the urethane hydrolase activity is 0.3U / ml.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses providencia for synthesizing ethanol-resistant urethanase and urease, and belongs to the technical field of bioengineering. A bacterial strain obtained by separation is identified as the providencia, and the synthesized urethanase and urease still have efficient enzyme activity under high-concentration ethanol (20 percent v / v) and acidic conditions (pH 4.5). Therefore, the application potential is huge in the removal of ethyl carbamate and urea in yellow wine and other fermented food, accordingly the control on the ethyl carbamate from a source to an end is realized, and the economical and social benefits are huge.

Description

technical field [0001] The invention relates to a Providencia strain for synthesizing ethanol-resistant urethane hydrolase and urease, belonging to the technical field of bioengineering. Background technique [0002] Ethyl Carbamate (Ethyl Carbamate or Urethane, referred to as EC), has genotoxicity and carcinogenicity, and is widely present in a variety of fermented foods (such as soy sauce, vinegar, pickles) and alcoholic beverages (such as rice wine, white wine, wine, etc.) . The International Agency for Research on Cancer listed urethane as a Group 2A carcinogen in 2007. The human body absorbs ethyl carbamate mainly through the consumption of alcoholic beverages and food. Urethane has become a non-negligible factor affecting human health. [0003] There are two main ways to eliminate EC in fermented food: one is to reduce or eliminate the content of EC precursors (mainly urea) in the fermentation broth, which is a common international practice. Urease breaks down urea...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/80A23L5/20C12R1/01
CPCC12N9/80C12Y305/01005C12Y305/01075C12N1/205C12R2001/01
Inventor 康振陈坚堵国成刘庆涛
Owner JIANGNAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com