A strain of Bacillus arborii producing neutral amylase

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

Active Publication Date: 2018-12-18
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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  • A strain of Bacillus arborii producing neutral amylase
  • A strain of Bacillus arborii producing neutral amylase
  • A strain of Bacillus arborii producing neutral 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 strain of bacillus arborii producing neutral amylase, which belongs to the technical field of bioengineering. The wild bacillus strain with high yield of neutral amylase is selected from shallow soil of cassava field in Guangxi. The strain of the present invention is preserved in the Chinese Type Culture Collection Center of Wuhan University, Wuhan City, Hubei Province, and its preservation number is CCTCC M 2017320. The culture conditions of this strain are simple, the fermentation period is short, and the enzyme activity is high; the amylase produced is a medium-temperature, neutral enzyme, and the optimum temperature and optimum pH distribution are 50°C and 7.0. It has a strong ability to hydrolyze raw starch, and the product is mainly maltose. . It has broad application prospects in industrial fields such as starch sugar, fermentation, food and feed.

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 Patents(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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