Bacterial strain LT3 producing alkalescence cellulase and breeding method and initial optimization of cellulase production conditions thereof

A cellulase and strain technology, applied in the field of microorganisms, can solve the problem of few reports on bacteria, and achieve the effect of rapid and accurate identification

Inactive Publication Date: 2009-10-14
FUZHOU UNIV
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  • Bacterial strain LT3 producing alkalescence cellulase and breeding method and initial optimization of cellulase production conditions thereof
  • Bacterial strain LT3 producing alkalescence cellulase and breeding method and initial optimization of cellulase production conditions thereof
  • Bacterial strain LT3 producing alkalescence cellulase and breeding method and initial optimization of cellulase production conditions thereof

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Embodiment Construction

[0016] The following are specific breeding methods of the present invention, and fully illustrate the present invention in conjunction with examples.

[0017] Breeding embodiment: 1 material and method

[0018] 1.1 Materials

[0019] Main reagents:

[0020] Sodium carboxymethyl cellulose, nitrosalicylic acid reagent (weigh 91g potassium sodium tartrate, dissolve in 500mL distilled water, heat to 50°C, then add 3,5-dinitrosalicylic acid (DNS) 3.5g , NaOH 20g, phenol 2.5g, anhydrous sodium sulfite 2.5g, stir to dissolve completely. After cooling, dilute to 1L with distilled water. Store in a brown bottle at 4°C, use after one week, filter before use.), PH4.6 acetic acid Buffer; the acetic acid buffer is configured by glacial acetic acid and sodium acetate, Congo red, anhydrous glucose, PCR mix enzyme (purchased from Beijing Tiangen Company), gel recovery kit (purchased from Dalian TaKaRa Company), and chemical reagents are all is analytically pure;

[0021] Medium (steriliza...

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Abstract

The invention provides a bacterial strain LT3 producing alkaline cellulose and a breeding method and an initial optimization of cellulose production conditions thereof. The bacterial strain LT3 with higher secretory volume of alkaline cellulose is obtained by steps of conducting accumulation culture, separation and purification, identification of Congo red dye and liquid fermentation on samples obtained from rotten wood or nearby soil thereof; the bacterial strain is identified to be bacillus cereus by initial identification, sequence clone and phylogenetic analysis; and 16s and r DNA sequences are shown as sequence list SEQ ID NO.1. The invention has potential theoretical and application values for research on the action mechanism of the alkaline cellulose and the production of the alkaline cellulose preparation; and the bacterial strain LT3 of the alkaline cellulose can produce the alkaline cellulose of single component and is convenient for separation and purification, thus being an ideal material for gene clone of the alkaline cellulose and having potential theoretical and practical significances for constructing alkaline cellulose fibrinolysis engineered bacterial strain and questing the action mechanism of the alkaline cellulose, and the method is simple, fast and easy for implementation.

Description

technical field [0001] The invention belongs to the technical field of microbes, and more specifically relates to an alkaline cellulase-producing strain LT3, a breeding method thereof, and preliminary optimization of enzyme-producing conditions. Background technique [0002] Cellulosic materials are the most abundant renewable resources in nature. The use of modern biotechnology to convert cellulose materials into liquid fuels such as ethanol can not only benefit human beings as new resources and new energy, alleviate or solve the problem of environmental pollution caused by fossil energy, but also realize the efficient use of agricultural surplus resources. It has been widely valued by countries all over the world. Using microbial technology to treat straw is currently the most researched straw treatment method. Cellulase can degrade natural cellulose and generate cellulose molecular chains, cellobiose and glucose. One of the key factors is the inefficiency of cellulase e...

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Application Information

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IPC IPC(8): C12N1/20C12Q1/68C12Q1/04C12R1/085
Inventor 吕暾李殿殿郑蓉黄爱玲吕静琳
Owner FUZHOU UNIV
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