Strain producing surface active agent resistance lipase and surface active agent resistance lipase preparation method

A surfactant and lipase technology, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, detergent compounding agents, etc. Properties, no problems such as lipase preparation methods, etc., to achieve the effect of enhanced reactivity

Active Publication Date: 2014-05-21
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0012] In addition, there are almost no reports about the lipase produced by Chryseobacterium bacteria, and there is no report on the enzymatic properties of Chryseobacterium lipase affected by any surfactant, and there is no report on the properties of Chryseobacterium lipase affected by surfactants. Report on the preparation method of lipase with surfactant tolerance from

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  • Strain producing surface active agent resistance lipase and surface active agent resistance lipase preparation method
  • Strain producing surface active agent resistance lipase and surface active agent resistance lipase preparation method
  • Strain producing surface active agent resistance lipase and surface active agent resistance lipase preparation method

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Embodiment 1

[0074] Embodiment 1, strain screening and identification

[0075] The bacterial strain Chryseobacterium WBRD1.11121501 provided by the present invention is isolated from the soil in Shanghai, China. A bacterial strain that can produce extracellular lipase was obtained by screening the newly collected soil samples on rhodamine B plates through enrichment culture or dilution coating. The strain was identified as Chryseobacterium sp. WBRD1.11121501 by 16SrRNA identification.

[0076] The strain has been deposited in the General Microorganism Center (CGMCC) of China Committee for Culture Collection of Microorganisms. The address of the depository unit is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard No. 1, Beichen West Road, Chaoyang District, Beijing, zip code 100101. The preservation date of the bacterial strain is June 21, 2012, the preservation number is CGMCC No.6261, and the classification name is: Chryseobacterium sp.

[0077] The screening...

Embodiment 2

[0083] Embodiment 2, preparation of surfactant-resistant lipase

[0084] Get a freshly cultivated Chryseobacterium aureus WBRD1.11121501 slant (LB culture medium 30 ℃ and cultivate 48h, connect a ring to the seed shake flask that contains 30ml seed culture medium, 30 ℃, 150rpm cultivate 24h. Then with 1% inoculum size Inoculate into the fermentation medium and culture on a shaker at 30°C and 150rpm for 48 hours. The culture solution is centrifuged or micro-filtered to remove bacteria, and the supernatant is concentrated by ultrafiltration using a 10kDa membrane. Then the concentrated solution is subjected to segmented ammonium sulfate salting out and collected 20-70% saturation of the salted-out precipitate. Then use the Hitrap phenyl HP chromatographic column (GE HealthCare, product number: 17-5195-01) to carry out hydrophobic chromatography purification on the salted-out protein sample, first use the starting buffer and The elution buffer carries out gradient elution, then e...

Embodiment 3

[0089] Embodiment 3, partial enzymatic property of lipase produced by Chryseobacterium WBRD1.11121501

[0090]The lipase prepared by the present invention has the following properties. Unless otherwise specified, the detection of lipase activity adopts the pNPP method (see the "Specific Embodiments" section).

[0091] Impact on Stability

[0092] The detection method that surfactant affects the stability of Chryseobacterium lipase is to get 2ml of the enzyme solution to be tested with an activity of about 0.3 units / ml, and add the surfactant to be detected until the surfactant concentration is 0.5% or 1.0 %(v / v), put it at room temperature, take out 0.1ml samples of some samples at the 20th, 45th and 70th minutes respectively for detection of lipase activity. The control enzyme activity is 100%, and the relative enzyme activity of the experimental sample is calculated.

[0093] The effect of surfactant on the stability of Chryseobacterium lipase is shown in figure 2 with ...

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Abstract

The invention discloses a strain WBRD1.11121501 producing surface active agent resistance lipase and a surface active agent resistance lipase preparation method using the strain. According to the invention, the lipase which is prepared by using the strain and the preparation method has strong stability in an anionic surface active agent SDS, nonionic surface active agents AEO-9 and Triton X-100 and a cationic surface active agent CTAB, and most of the surface active agents can enhance the reaction activity of the lipase. In addition, oxidant hydrogen peroxide can enhance the reaction activity of chryseobacterium lipase provided by the invention.

Description

technical field [0001] The invention relates to a strain producing lipase and a preparation method of lipase. More specifically, the present invention relates to a strain producing surfactant-resistant lipase and a method for preparing surfactant-resistant lipase using the strain. Background technique [0002] Lipase (lipase, EC3.1.1.3) is a special class of ester bond hydrolase, which widely exists in animals, plants and microorganisms. Lipase can catalyze the hydrolysis, alcoholysis, esterification, transesterification and compound synthesis of ester compounds, which can be used in food, medicine, detergent, textile, biodiesel, papermaking, leather, cosmetics and environmental protection, etc. Various industrial fields (Hasan F, Ali SA, Hameed A. Industrial applications of microbial lipases. Enzyme Microb. Technol. 2006, 39: 235-251.). [0003] Lipase, like other industrial enzyme preparations, has a variety of properties in different application fields. [0004] Becaus...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/20C12P7/64C11D3/386C12R1/01
Inventor 徐正军周美凤许骏柴丹
Owner WILMAR SHANGHAI BIOTECH RES & DEV CENT
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