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Bacillus addisoni capable of producing neutral raw starch digesting amylase

A technology of Bacillus arborii and raw starch, applied in the field of bioengineering, can solve the problems of insufficient activity of bacteria, poor enzymolysis or fermentation effect, loss of enzyme activity, etc., and achieve the effect of strong ability to hydrolyze raw starch

Active Publication Date: 2018-03-06
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The pH is not the best condition for either party, so it will cause loss of enzyme activity or insufficient bacterial activity, resulting in poor enzymatic hydrolysis or fermentation

Method used

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  • Bacillus addisoni capable of producing neutral raw starch digesting amylase
  • Bacillus addisoni capable of producing neutral raw starch digesting amylase
  • Bacillus addisoni capable of producing neutral raw starch digesting amylase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 produces the screening of amylase strain

[0044] Soil samples were collected from 10-20cm shallow soil of cassava fields in Wuming District and Suining District, Nanning City, Guangxi, and a total of 16 soil samples were collected. Weigh 2g of soil sample into a 250mL Erlenmeyer flask, add 20mL of sterile saline. Stir on a magnetic stirrer for about 30 minutes, take the suspension and heat-treat at 80°C for 15 minutes to kill the vegetative cells, and then enrich and culture at 37°C for 24 hours. Then take the enriched culture solution for gradient dilution (10 3 、10 4 、10 5 and 10 6 ), 50 μL each was spread on a screening medium plate with tapioca starch as the sole carbon source, and cultured at 30°C for 2 days. Observe the growth of the colony, and add 0.05% dilute iodine solution dropwise, measure the diameter (D) of the transparent circle around the colony and the diameter (d) of the colony, and calculate their ratio.

[0045] Select 58 colonies...

Embodiment 2

[0046] Embodiment 2 produces the identification of amylase strain

[0047] (1) Colony characteristics and cell morphology

[0048] The colonies on the solid screening plate are round and regular, with smooth edges, raised surface, milky white, moist, opaque, easy to provoke, and wrinkle-free ( figure 1 ). It is white waxy when observed against light. The colonies will be slightly yellow if cultured for a long time. Microscopically, the bacteria were long rod-shaped, with spores, and Gram staining was positive.

[0049] (2) Physiological and biochemical characteristics

[0050] The starch hydrolysis test was positive. Catalase, oxidase, urease, galactosidase, and phosphatase tests were positive. Trypsin, oxidase (Kovacs), aromatase, and indole tests were negative. The methyl red test was negative. Can utilize glucose, xylose, arabinose, mannose, raffinose. Lactose, acetate, sorbitol, rhamnose cannot be utilized.

[0051] (3) 16S rDNA sequence analysis

[0052] Inocul...

Embodiment 3

[0056] The influence of embodiment 3 different carbon sources on thalline growth and enzyme production

[0057] The preserved Bacillus arzii was inoculated into the seed medium, and cultured in a shaker flask at 37° C. and 200 rpm for 12 hours as seeds. The seeds were inoculated into the fermentation medium with an inoculation amount of 2.5% (V / V), and the shaker was shaken at a rotational speed of 200 r / min.

[0058] Tapioca starch, corn starch, potato starch, glucose, maltose, fructose, glycerol, and sucrose were used as carbon sources for the fermentation of strain GEL-09, and other medium components and conditions were unchanged. The bacterial concentration and enzyme activity were measured after fermentation. The result is as image 3 As shown in A, when different types of starch are used as carbon sources, the growth of bacteria is low, but the activity of enzymes produced is high; when monosaccharides, disaccharides, and glycerol are used as carbon sources, although th...

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Abstract

The invention discloses a bacillus addisoni capable of producing neutral raw starch digesting amylase, belonging to the technical field of bioengineering. The wild bacillus strain capable of high-yield raw starch digesting amylase is selected from Guangxi cassava field shallow soil. The strain is preserved in China Center for Type Culture Collection in Wuhan university in Hubei province, with collection Number of being CCTCC M 2017320. The strain is simple and convenient in culturing conditions, short in fermentation period, high in enzyme activity; the produced raw starch digesting amylase isintermediate-temperature neutral enzyme, the most proper temperature and most proper pH distribution are respectively 50 DEG C and 7.0, the raw starch hydrolyzing capability is strong, and the product is mainly maltose. The bacillus addisoni has wide application prospects in the industrial fields of starch sugar, fermentation, food, feed and the like.

Description

technical field [0001] The invention relates to a strain of bacillus arborii producing neutral amylase, which belongs to the technical field of bioengineering. Background technique [0002] Starch is one of the most abundant carbohydrates in nature. It is not only the main source of nutrition for humans and animals, but also an important industrial raw material. Starch can be used not only to produce starch sugars such as glucose and maltose, but also to produce fermented products such as amino acids, organic acids, liquor, beer, alcohol, and vinegar. Starch is a polysaccharide composed of glucose as a unit. In industrial applications, it first needs to be enzymatically hydrolyzed into monosaccharides or oligosaccharides. According to whether the starch substrate used for enzymatic hydrolysis has been gelatinized, the enzymatic hydrolysis process is generally divided into two categories: high-temperature cooking process and raw meal enzymatic hydrolysis process. In the hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/26A23L33/135A23K10/16C12R1/07
CPCA23K10/16A23L33/135C12N9/2411C12N1/205C12R2001/07
Inventor 段绪果沈镇炎周素雅陶峥赵娅如张钰
Owner NANJING FORESTRY UNIV
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