A kind of strain producing surfactant-resistant lipase and preparation method of surfactant-resistant lipase

A surfactant and lipase technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, detergent compounding agents, etc., can solve the problem of no lipase produced by bacteria of the genus Chryseobacterium, no lipase preparation method, and the impact Problems such as the enzymatic properties of Chrysobacterium aureus lipase achieve the effect of enhancing the reactivity

Active Publication Date: 2018-01-12
WILMAR SHANGHAI BIOTECH RES & DEV CENT
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  • Abstract
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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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  • A kind of strain producing surfactant-resistant lipase and preparation method of surfactant-resistant lipase
  • A kind of strain producing surfactant-resistant lipase and preparation method of surfactant-resistant lipase
  • A kind of strain producing surfactant-resistant lipase and preparation method of surfactant-resistant lipase

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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 and ...

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Abstract

The invention discloses a strain WBRD1.11121501 producing surfactant-resistant lipase and a preparation method of the surfactant-resistant lipase using the strain. The lipase prepared by utilizing the bacterial strain provided by the present invention and the preparation method has extremely strong stability in anionic surfactant SDS, nonionic surfactant AEO‑9, Triton X‑100 and cationic surfactant CTAB, and wherein Most surfactants enhance the reactivity of the enzyme. In addition, the oxidant hydrogen peroxide also enhances the reactivity of the Chrysobacterium aureus lipase provided by the present invention to a certain extent.

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 Patents(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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