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Bacillus licheniformis and method for producing alkali proteinase with sewage sludge as raw material

A technology of bacillus licheniformis and protease, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems of undiscovered production of alkaline protease, expensive raw materials, high product prices, etc., to promote industrial production and Application, environment safe and harmless, high added value effect

Active Publication Date: 2010-06-02
山西卓奇水务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the mainstream production process of alkaline protease is liquid submerged fermentation, generally using soybean powder, starch, minerals, etc. as fermentation raw materials, these raw materials are expensive, resulting in higher product prices
So far, no report on the production of alkaline protease using municipal sludge as fermentation raw material has been found.

Method used

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  • Bacillus licheniformis and method for producing alkali proteinase with sewage sludge as raw material
  • Bacillus licheniformis and method for producing alkali proteinase with sewage sludge as raw material
  • Bacillus licheniformis and method for producing alkali proteinase with sewage sludge as raw material

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Isolation and purification of bacterial classification

[0035] Weigh 10 grams of concentrated sludge into a conical flask containing 90 mL of sterile water (with glass beads inside), shake and cultivate for 15 minutes at 28°C, 150 rpm, and dilute to 10 with sterile water. -5 , pipette 0.1mL of the diluted solution and evenly spread it on a casein medium plate, and cultivate it in a 28°C incubator for 3 days. Spread trichloroacetic acid solution evenly on the medium plate, pick single colonies with obvious hydrolysis circles and culture them repeatedly on the casein plate, and then separate, purify and store them. The ingredients of the casein medium used are: 20 g of casein, 20 g of agar powder, pH 9.0, and 1 L of distilled water. The slant preservation medium used was LB medium, and its composition was (g / L): 10 g of tryptone, 5 g of yeast extract, 10 g of NaCl, pH 7.4, and 1 L of distilled water.

Embodiment 2

[0036] Embodiment 2. strain identification

[0037] (1) Physiological and biochemical characteristics identification

[0038] See Table 1 for the identification results of physiological and biochemical characteristics of B.licheniformis HM-3.

[0039] Table 1. Physiological and biochemical characteristics of strain HM-3

[0040]

[0041] Note: + indicates that it can be used; - indicates that it cannot be used

[0042] (2) BIOLOG identification

[0043] The identification result of automatic microbial identification instrument (BIOLOG) showed that HM-3 was Bacillus licheniformis. The scanning electron microscope photo of Bacillus licheniformis HM-3 is shown in figure 1 .

Embodiment 3

[0044] Embodiment 3. bacillus licheniformis HM-3 takes sludge as raw material fermentation production alkaline protease

[0045] (1) Medium conditioning: adjust the solid content of urban sludge to 3%, add 0.2% foam enemy, and adjust the pH to 7.0 with NaOH, stir evenly, sterilize at 0.1KPa pressure and 121°C for 45 minutes, cool for later use.

[0046] (2) Shake flask culture: Pick a ring of bacterial lawn from the HM-3 slant medium and inoculate it into a conical flask filled with LB liquid medium, and carry out shake flask culture at 30°C and 100 rpm for 12 hours.

[0047](3) Fermentation: With the inoculum amount of 3%, the shake flask bacterial liquid was moved into a fermenter equipped with sludge with a solid content rate of 3%, and fermented. The conditions are: tank temperature 30°C, tank pressure 0.005 MPa, ventilation rate 1:1.0 (V / V / min), stirring speed 200 rpm, fermentation time 64 hours.

[0048] (4) Purification of protease: after the fermentation is completed,...

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Abstract

The invention discloses a bacillus licheniformis and a method for utilizing the sludge as raw material for producing alkaline protease. The bacillus licheniformis is gram-positive bacteria and is shaped like a straight rod, and the collection number is CGMCC 1970. The method for utilizing the bacillus licheniformis for producing the alkaline protease includes (1) the conditioning of a culture medium; (2) shaking culture; (3) the fermentation by a fermentation tank; and (4) the purification of the protease. The bacterial strain of the invention is utilized for producing the alkaline protease, which can not only dispose sludge, but also obtain the alkaline protease product with high additional value and lower cost. The method uses urban waste sludge to replace the traditional food culture medium for producing the alkaline protease product, which not only provides a new way for the resource disposal of urban sludge, but also can greatly reduce the price of the product and promote the industrial production and application of protease preparations.

Description

technical field [0001] The invention relates to the fields of new strain screening and enzyme preparation production, in particular to a bacillus licheniformis and a method for producing alkaline protease using urban sludge as a fermentation raw material. Background technique [0002] Municipal sludge is a sediment produced in the process of sewage treatment. It contains a large amount of organic matter, rich in nitrogen and phosphorus, and pathogenic bacteria. It is an important source of environmental pollution. At present, the annual output of urban sludge in my country is about 8 million tons of dry weight, and with the increase of sewage treatment plants and the improvement of sewage treatment rate, the output will become increasingly large. How to properly dispose of sewage sludge, prevent its secondary pollution, and use it as a useful resource to turn waste into treasure is of great significance. [0003] At present, the disposal methods of municipal sludge mainly i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/56C12R1/10
Inventor 周顺桂王跃强雷发懋
Owner 山西卓奇水务有限公司
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