Alkali-fast bacillus cereus bacterial strain and produced pectinase thereof

A technology of bacillus and pectinase, applied in the field of bioengineering, can solve the problems of low thermal stability, low degree of industrial production, and high cost of enzyme production, and achieve the effects of simple application and operation, avoiding inhibition, and good repeatability

Inactive Publication Date: 2011-04-06
CHANGSHU INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Yet, because alkaline pectinase high-yielding bacterium is few, the cost of producing enzyme is higher, thermostability is lower, and its industrialized production degree is still very low, therefore, breeding high-yielding bacterial strain improves its thermostability, alkali stability, Improving the yield of alkaline pectinase and reducing the cost of enzyme production are the key to the research, development and application of alkaline pectinase

Method used

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  • Alkali-fast bacillus cereus bacterial strain and produced pectinase thereof
  • Alkali-fast bacillus cereus bacterial strain and produced pectinase thereof
  • Alkali-fast bacillus cereus bacterial strain and produced pectinase thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0032] This example illustrates the procedures for the screening and identification of Bacillus alcalophilus M29.

[0033] Screening medium (g L -1 ): pectin 30, yeast paste 1, ammonium nitrate 3, potassium dihydrogen phosphate 6, sodium carbonate 10 (sterilized separately), magnesium sulfate heptahydrate 0.5, pH 10.0;

[0034] Screening plate medium (g L -1 ): pectin 20, peptone 5, yeast extract 3, sodium carbonate 10 (sterilized separately), agar 20, pH 10.0;

[0035] Fermentation medium (g L -1 ): pectin 5, sucrose 20, yeast extract 1, sodium carbonate 10 (sterilized separately), corn steep liquor 20mL (dry weight about 4g), potassium dihydrogen phosphate 6, magnesium sulfate 0.5, pH 10.0;

[0036] Broth plate medium (g L -1 ): peptone 10, beef extract 5, sodium chloride 5, agar 20, pH 10.0.

[0037] 1. Primary screening:

[0038] About 1,500 soil samples were randomly collected from various places in Jiangsu Province, and about 1 g of each of 3 to 4 kinds of soil sam...

Embodiment 2

[0044] This experiment illustrates the morphological characteristics of Bacillus alcalophilus M29.

[0045] The present invention identifies the biological characteristics of Bacillus fordii MH602, which is Gram-positive bacteria with spore formation. After growing in broth plate medium for 16 hours, the colony size is 2-2.5 mm, the growth temperature range is 45-55 °C, the optimum temperature is 50 °C, the growth pH is 8.0-11, and the optimum pH is 10. Physiological and biochemical manifestations are positive peroxidase test and growth under aerobic conditions.

Embodiment 3

[0047] This example illustrates the thermostability of pectinase produced by Bacillus alcalophilus M29.

[0048] Fermentation method of pectinase:

[0049] Fermentation medium (g / L): 5 pectin, 20 sucrose, 1 yeast extract, 10 sodium carbonate (sterilized separately), 20 mL corn steep liquor (about 4 g in dry weight), 6 potassium dihydrogen phosphate, 0.5 magnesium sulfate, pH 10 .0. Sterilize at 121°C for 20 minutes. Inoculate the strains on the broth medium into the fermentation medium, ferment at 55°C and 200rpm for 18-22h, then centrifuge at 8000rpm for 10min, and the supernatant is the pectinase crude enzyme solution.

[0050] Divide the pectinase solution into different reaction tubes, incubate at 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C for one hour, then take 20 μL for different time Add substrate 20g·L to the pectinase solution after heat preservation -1 Polygalacturonic acid was converted at 55°C for 10 min, and the reaction was terminated with 3 mL of ...

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Abstract

The invention uses high-concentration pectin as a unique carbon source to screen an alkaline-resisting bacillus strain from soil; the strain is named Bacillus alcalophilus M29, the preservation date is January 6th, 2009, the full name of the preservation institution is China Center for Type Culture Collection, and the preservation number is CCTCC NO: M 209006; the invention further relates to pectase produced by alkaline-resisting Bacillus alcalophilus M29, having better thermal stability (45 to 65 DEG C), substrate tolerance and basophilia (8.5 to 11, the optimum pH value being 10) and belonging to heat-resisting basophilia pectase; and the invention also relates to the application of the alkaline-resisting Bacillus alcalophilus M29 strain in fermentation and preparation of pectase.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to an alkali-resistant bacillus Bacillus alcalophilus M29 with high production of thermostable alkalophilus pectinase and the thermostable alkalophilus pectinase produced therefrom. Application of glue enzymes. Background technique [0002] Pectinase refers to a type of enzyme that can decompose pectin, and alkaline pectinase (E.C.4.2.2.2) refers to a type of pectinase with relatively high activity in the alkaline range. Since Japanese scholar Koki Horikoshi made alkaline pectinase with alkaliphilic bacteria for the first time in 1972 (Hoondal G.S., et al. Appl Microbiol Biotechnol, 2002, 59: 409-418), its application research has become more and more extensive. In addition to the application of alkaline pectinase in the purification of plant viruses and pulp bleaching, it has become a vital biological enzyme preparation in the pretreatment of cotton fabrics. Th...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/24C12R1/07
Inventor 齐斌梅艳珍王立梅郑丽雪
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
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